Eggshell color is an important intuitive economic trait and attracts much attention for a long time. In China, uniform color eggs gain more popularity among consumers. However, different chicken breeds produced eggs with different eggshell color, and even in the same breed, there is difference in eggshell color among different individuals. Thus, it is inevitable to improve the uniformity of eggshell color for breeders. To reveal the determinants for eggshell color at the molecular level, this review elucidated the basic property of eggshell color and discussed its genetic patterns, genes and QTLs involved in eggshell color regulation, which would provide reference for designing breeding goals and breeding program.

Automation and information are the important development directions of technologies in dairy industry. To break through the bottleneck of automatically monitoring technology for quiet estrus cows, the regulation pattern of body temperature has become another study focus related to reproductive cycle of dairy cattle after the regulation pattern of activity quantity. Both age and diurnal variation characteristics of body temperature in cow have been shown. Meanwhile, the regulation pattern of body temperature related to estrus, pregnancy and delivery on reproductive cows was observed, and the regulation patterns varied with different measurement sites. Among these detection sites, the regulation pattern of viginal temperature plays a more instructive role in marking reproductive behavior for its close relationship to reproductive activities. Additionally, the body temperature was usually affected by reproductive hormones, environmental conditions, diseases, measurement methods and other factors. The premises of making better use of body temperature data are scientific analysis of factors affecting body temperature and construction of an automatic collection technology and database for it, which will promote the automation, elaboration and intelligentization of both cow reproduction and health management.

The present study aimed to explain the domestication regions of Chinese chicken and its genetic propagation by analyzing the distribution and frequencies of D-loop haplotype clades of domestic chicken and wild red jungle fowls. The results showed that 198 haplotypes gained by sequencing 1 347 mtDNA D-loop fragments were clustered into 12 clades. Cluster analysis indicated that the island-type red jungle fowls (G. g. bankiva) were significantly different from domestic chicken and continental red jungle fowls (G. g. gallouse) of Southeast Asian, and that continental red jungle fowls (G. g. spdiceus) in Sumatra Island were independently branched from those of Southeast Asian. The distribution and frequencies analysis of haplotype clades indicated that the early domestic chicken with maternal blood presented by Clade C could origin from Southeast Asian and its surroundings(including Yunnan), but was not directly introduced into China via Yunnan. The maternal blood presented by Clade D could be domesticated in Shandong, Henan and its surroundings at early age, and then was introduced into Sichuan. From then on, the maternal blood began to reproduce on a large scale, propagated to surroundings, and became an important genetic source of Chinese domestic chicken. The present study can be an additional evidence to support the inference that Chinese domestic chicken have multiple maternal origins, and also indicate that Southeast Asian is an important original region of chickens in South China. In addition, Henan, Shandong and its surroundings are also important domestication regions of Chinese chicken.

The study aimed to investigate the mutation of GNAS gene and its effect on skin color traits in chicken. Dongxiang Blue-eggshell chicken (Gallus gallus domestica) was chose and the GNAS mutation sites related to skin color were screened using PCR-restriction length polymorphism (RFLP) and DNA sequencing in this study. We detected expression of the EDN3, MC1R, GNAS and ADCY3 genes using fluorescence quantitative PCR, and the mutation sites among 3 chicken breeds. The results showed that there was a T/C mutation at base 11 167 660 on chicken chromosome 20 and the mutation (T2270C) was crucial for the GNAS gene promoter. After Dra I enzyme digestion for PCR products, the CC homozygous genotype, the TC heterozygous genotype and the TT recessive homozygous genotype presented one band at 342 bp, two bands at 171 and 342 bp, and one band at 171 bp, respectively. The L values of skin color were statistically significantly different among 3 genotypes (P＜0.01). The individuals with CC genotype had the lowest L value, which presented the darkest skin, and the individuals with TT genotype had the highest L value, which presented the lightest skin. Expression of the EDN3 gene was highest and lowest in individuals with the CC and TT genotype, respectively, and the difference among 3 genotypes was extremely significant (P＜0.01). Expression of the MC1R, GNAS and ADCY3 genes was highest in individuals with CC genotype, but the difference among genotypes was not significant (P＞0.05). The correlation of GNAS with MC1R was non-significant in expression, and the correlation with ADCY3 was extremely significant (P＜0.01). The typing results according to mutation detection in Jiangshan Black-bone chicken, ISA layer, test-crossing offspring of Dongxiang Blue-eggshell cock and ISA hen, conformed to the theoretical values (P＞0.05). The study indicate that the T2270C mutation in GNAS gene promoter in chicken is correlated strongly with the skin color traits, and detection of the mutation can be used in the breeding of black-bone chicken.

The objectives of the present study were to investigate the characteristics of expression and regulation of CYP2C8 gene in chicken. A total of 14 tissues (including heart, liver, spleen, lung, kidney, pectoral muscle, abdominal fat, gizzard, glandular stomach, ovaries, pancreas, duodenum, jejunum and ileum) were collected from Lushi green-shelled-egg chickens at the age of 30 weeks old, and the liver tissues were obtained from the chickens at different development stages (from the age of 1 day to 30 weeks old) . The samples were used to detect the spatiotemporal expression profiles of CYP2C8 gene using quantitative real-time PCR (qRT-PCR) technology. In addition, the pullets (at the age of 10 weeks old) which were treated with 17β-estradiol, and chicken embryo primary hepatocytes which were treated with either 17β-estradiol alone or combination of 17β-estradiol and estrogen receptor (ER) antagonists were used to analysis the expression regulation mechanism of the CYP2C8 gene. The results showed that CYP2C8 gene was expressed extensively in different tissues of chicken. The relative expression level of CYP2C8 was higher in the main lipid metabolism tissues such as liver, kidney, and small intestine (duodenum, jejunum and ileum) than that in other tissues. The expression level of CYP2C8 gene in liver of hens was significantly increased with the sexual maturity (P<0.01). Estrogen could significantly up-regulate the expression of CYP2C8 in liver of chicken and hepatocytes (P<0.05). MPP, as an ER-α specific antagonist, could completely antagonize the expression of CYP2C8 gene (P<0.05). Therefore we concluded that CYP2C8 gene was expressed extensively in different tissues with higher expression levels in liver, kidney, and small intestine (duodenum, jejunum, and ileum) tissues, and the highest expression level in liver. It was suggested that the effect of estrogen on the expression of CYP2C8 gene was predominantly mediated via ER-α in the chicken liver.

To understand the molecular regulation differences of muscle growth and development in purebred and crossbred pigs, and analyze its effect on carcass and meat quality in pig. Duroc, Landrace, Large White pigs and Duroc×(Landrace×Large White) crossbred pigs with the average body weight of 50 kg were used. Each group was repeated by 6 individuals, and longissimus dorsi muscle was collected and analyzed at both phenotypic and molecular levels. The results showed that, compared with the purebred pigs, crossbred pigs had significant higher myofibre cross-sectional area, white myofibre ratio, lactate dehydrogenase A gene expression level and mtDNA copy number (P<0.01); On the whole, the expression level of IGF1, PDK1 and GLUT4 genes promoting myofibre growth and development in crossbred pigs were significantly increased (P<0.05), the expression level of MSTN and FOXO1 genes inhibiting myofibre growth and development in crossbred pigs were significantly decreased (P<0.05); the expression level of PGC1A gene promoting white myofibre conversion in crossbred pigs was significantly increased (P<0.05), the expression level of MEF2A, MYOZ1, MRF4 and NFATC1 genes inhibiting white myofibre conversion in crossbred pigs were significantly decreased (P<0.05); the expression level of miR-1 and miR-133 related to myofibre proliferation in crossbred pigs were significantly increased (P<0.01), while the expression level of miR-499 inhibiting white myofibre growth in crossbred pigs was significantly decreased (P<0.01). These results suggest that improving carcass traits in pigs through corssbreeding is achieved by changing its molecular regulation level, which had adverse effects on meat quality.

The objectives of this study were to investigate the developmental changes of FAM134B, PPARγ, FAS and HSL gene expression in the process of sheep muscles development and their association with intramuscular fat (IMF) accumulation, to lay a theoretical foundation for studying molecular regulation mechanism of the IMF deposition in sheep. Five Aohan fine wool sheep were slaughtered at 2, 4, 5, 6 and 12 months old to collect samples from longissimus dorsi and biceps femoris, for the purpose of determining the IMF content. Then mRNA expression level of the FAM134B, PPARγ, FAS and HSL gene in different development period of muscles were analyzed by real-time PCR, and association between genes expression and IMF contents were analyzed. The results showed that: (1) Among 2-5 months old, with growing of the age, the IMF contents of longissimus dorsi and biceps femoris were increasing. Among 5-12 months old, the IMF contents remained unchanged; The IMF content of longissimus dorsi were extremely significantly higher than biceps femoris (P<0.01) at the same age. (2) In longissimus dorsi, FAM134B expression level presented first increased and then declined with growing, the highest was in 6 months old(P<0.01). In biceps femoris,FAM134B expression level presented increased and then stabilized with growing. FAS expression level increased with growing in both of two parts of muscles. PPARγ expression level presented first declined and then increased with growing. HSL expression levels was showed “declined-increased-declined” patterns in two parts of muscles. In the same month, target genes expression level had tissue-dependent in two parts of muscles.(3) Correlation analysis showed that the expression level of FAM134B was significantly positively related to FAS in two parts of muscles(P<0.05), and was negatively related to PPARγ and HSL expression levels at different levels. In longissimus dorsi, FAM134B, FAS expression levels were significantly positively related to IMF content(P<0.01 or P<0.05, respectively), PPARγ expression level was significantly negatively related to IMF content(P<0.05). In biceps femoris, FAM134B expression level was significantly positively correlated with the IMF content(P<0.05). In conclusion, FAM134B, FAS expression might have positively effect on IMF accumulation, and PPARγ, HSL expression might have negatively effect on IMF accumulation. FAM134B was likely to regulate IMF content through stimulating expression level of fat synthetic genes and inhibiting adipose decompose genes. The results would provide reference for studying related gene functions and molecular mechanism of the IMF deposition.

In order to understand the expression and cellular localization of double sex and mab-3 related transcription factor 7 (Dmrt7) in Small-Tail Han sheep testis tissues, fifteen sheep testis tissues were collected at 0, 2, 6, 12 and 24-month-old stages (3 replicates each), and the expression and cellular localization of Dmrt7 on testis tissues were detected by qRT-PCR, Western blot and immunohistochemistry. The results showed that Dmrt7 mRNA and protein were detected in sheep testes at different months of age, and the change pattern was similar. But, there were differences concerning the expression level of Dmrt7 for the different month groups. Dmrt7 mRNA and protein were lowly expressed in 0, 2 and 6 -month-old sheep testes, but it were highly expressed in 12, 24-month-old sheep testes. Meanwhile, the expression of Dmrt7 mRNA and protein in 12 and 24-month-old sheep testis tissues were extremely significantly higher than those in 0, 2 and 6-month-old sheep testes tissues（P<0.01）. The immunohistochemically staining revealed that immunoreactivity of Dmrt7 protein was localized to minority of spermatogonia in 0, 2 and 6-month-old sheep testis tissues, but it was mainly localized to secondary spermatocytes and partial primary spermatocytes and spermatids in 12 and 24-month-old sheep testis tissues. The results demonstrated that Dmrt7 played an essential regulation role to the division processes of various spermatogenic cells in testicular seminiferous tubule, especially meiotic division from primary spermatocytes to spermatids.

The aim of this study was to construct a dual luciferase reporter plasmid containing the 3′untranslated region(UTR) of bta-GHR, and verify the targeting relationship between miR-139 and bta-GHR. The binding site of miR-139 to bta-GHR were predicted using bioinformatics. The synthetic 3′UTR fragment of bta-GHR was cloned into psiCHECK-2 vector to construct wild type (psiCHECK-2-GHR-W 3′UTR) or mutant type (psiCHECK2-GHR-M 3′UTR) dual luciferase reporter plasmid. Cultured Hela cells were divided into 4 groups, they were co-transfected with miR-139 mimic or negative control and psiCHECK2-GHR-W 3′UTR or psiCHECK2-GHR-M 3′UTR. The relative luciferase activity was detected. The cultured bovine mammary epithelial cells were transfected with miR-139 mimic, negative control or miR-139 inhibitor. Using real-time quantitative PCR (qPCR), we measured the mRNA level of GHR and miR-139 in cow. The results showed that, by digestion of restriction enzymes, the wild type recombinant plasmid (psiCHECK2-GHR-W 3′UTR) and mutant type recombinant plasmid (psiCHECK2-GHR-M 3′UTR) were proved to be successfully constructed. After psiCHECK2-GHR-W 3′UTR and miR-139 mimic co-transfected Hela cells, the luciferase activity was significantly lower than other treatment groups (P<0.05). Using qPCR,we further confirmed that the mRNA level of GHR was down-regulated by miR-139 (P<0.05). The dual luciferase reporter plasmid containing 3′UTR of wild type or mutant type of bta-GHR was constructed successfully; The targeting relationship between miR-139 and bta-GHR was confirmed by dual luciferase assay and qPCR assay.

On the basis of previous results, we transformed a bovine Desmin promoter fragment to obtain some high efficiency and muscle tissue-specific promoters which could be useful. We obtained 3 artificial Desmin promoter fragments successfully through adding several positive specific regulatory elements into the bovine Desmin promoter of 300 bp length, which included pGL3-desmin300-desmin180, pGL3-MD-αD①-desmin300 and pGL3-αD②-αD①-desmin300. The results showed that the activities of pGL3-desmin300-desmin180, pGL3-MD-αD①-desmin300 and pGL3-αD②-αD①-desmin300 were 1.67, 2.56 and 2.88 times more than activity of pGL3-desmin300, respectively. Bovine skeletal muscle satellite cells and fibroblast cell experiments showed that the 3 artificial Desmin promoter fragments had high muscle specificity. The application of Desmin fragments with high efficiency and muscle specificity will provide an important help to increase the output of muscle production of transgenic livestock.

To compare and analyze the differences and similarities between two versions of Code of Practice of Type Classification in Chinese Holstein (use Code of Practice for short) , a total of 2 174 first lactation Holstein cows from 26 farms in Ningxia area were evaluated by 9-points type classification system. The weights from the two versions of Code of Practice were used to evaluate the score of 5 composites (Body size, rump, feet and legs, udder, dairy form) and type final score. The results showed that the weight of Udder was increased while the weight of Dairy Form was decreased. Compared with the final score derived from the old version, the range of final score from the new version was wider, which can explore the difference more efficiently among individuals within the population and difference among daughters from different bulls, respectively. The results conclude that the new version of Code of Practice can better fit to Chinese Holstein, and more beneficial to promote the genetic gain.

The objective of this study was to investigate the effects of long-term high-grain (HG) feeding on rumen environment, lipopolysaccharide concentration in jugular vein blood, and the occurrence of subacute ruminal acidosis in goats. Ten castrated male goats with rumen fistula were randomly fed with either a hay diet (0% grain; n = 5) or HG diet (65% grain; n = 5) with continuous feeding of 7 weeks. Weekly 0, 2, 3, 4, 6, 8 and 12 h after morning feeding, rumen fluid was collected to monitor the changes of pH in rumen. The rumen fluid collected at 0, 3, 6 and 12 h after morning feeding was used to determine volatile fatty acid concentration. The rumen fluid collected at 6 h after morning feeding was used to determine lipopolysaccharide concentration. At 6 h after morning feeding, the jugular vein blood was collected to measure the free lipopolysaccharide levels in peripheral blood. The results showed that, at 1st, 2nd, 3rd, 6th and 7th weeks, the rumen pH below 5.8 lasted for more than 4 h within 12 h after morning feeding in HG-fed goats. The results suggested that, at 1^st, 2^nd, 3^rd, 6^th and 7^th weeks, the experimental subacute ruminal acidosis had been successfully induced by HG. Across the whole period, compared with hay feeding, HG feeding significantly decreased (P<0.05) the ruminal pH, acetate proportion, and acetate/propionate ratio, while significantly increased (P<0.05) ruminal propionate proportion, butyrate proportion, and free lipopolysaccharide concentration in rumen. The free lipopolysaccharide in the peripheral blood was not detectable in all hay-fed goats across the whole period. At the 1^st and 2^nd weeks, the free lipopolysaccharide level in the peripheral blood of HG-fed goats was not detectable. At 3^rd and 4^th weeks, the free lipopolysaccharide in the peripheral blood was detectable in some HG-fed individuals. At 5^th, 6^th and 7^th weeks, the free lipopolysaccharide in the peripheral blood were detectable in all HG-fed goats. These results indicate that experimental subacute ruminal acidosis has been successfully induced by long-term HG diet feeding, and the incidence of subacute ruminal acidosis experienced the process of occurrence, adaptation and repeated occurrence.

This experiment was conducted to estimate distributions of Cu, Fe, Mn, Zn and the net requirements for maintenance and growth of Dorper × Jinzhong crossbred ram lambs at 35-50 kg. A total of 30 ram lambs were used in this experiment. A comparative slaughter trial was carried out, and 6 lambs were randomly chosen and slaughtered at approximately 35 kg BW as the baseline group for measuring initial body composition. Another 6 lambs were also randomly chosen and offered a pelleted mixed diet for ad libitum intake and slaughtered at approximately 43 kg BW. The remaining 18 lambs were equally assigned to 3 groups in a randomised design according to the level of dry matter intake (ad libitum or 65% or 40% of ad libitum intake of the same diet), the 3 groups were slaughtered when the sheep fed ad libitum reached approximately 50 kg BW. The contents of Cu, Fe, Mn and Zn in different tissues, including muscle, adipose, bone, blood+visceral, leather and wool, were measured; mathematical model for analyzing distributions of trace element in tissues and predicting trace element requirements for maintenance and growth were established. The results showed that the growth rate of bone, muscle and fat decreased, decreased and increased, respectively. Cu, Fe Mn were mainly stored in viscera, accounting for 86.34%, 49.12%, 68.65% of the total contents in body, while Zn was mainly stored in muscle and bone, accounting for 42.17%, 24.19% of the total contents in body. The net requirements for maintenance of Cu, Fe, Mn, Zn expressed as empty body weight (EBW) were 0.016, 0.281, 0.007 and 0.085 mg•kg^-1 EBW•d^-1, and expressed as body weight (BW) were 0.013, 0.228, 0.006 and 0.069 mg•kg^-1 BW•d^-1, respectively. Net requirements for growth expressed as empty body weight (EBW) were 12.61-15.62, 77.82-82.55, 2.74-3.45 and 25.36-23.98 mg•kg^-1 EBW, and expressed as body weight (BW) were 10.17-12.75, 62.76-67.39, 2.21-2.82 and 20.45-19.57 mg•kg^-1 BW, respectively. The net requirements for maintenance of trace element in this study were higher than that recommended by NRC, excepted for Zn; net requirements for growth of Fe and Zn were similar with the recommendation of NRC, while requirements of Cu and Mn were higher than the recommendation.

The objective of this study was to investigate the effects of starter age on Hu lamb genes expression involved in cellular growth in rumen papilla and rumen morphology.Two factor design (starter age and age of lambs) was adopted in this study.A total of 78 male Hu lambs with similar born body weight （(3.41±0.27) kg） were selected.Of which，six were slaughtered immediately after birth and the remaining 72 were equally divided into 2 groups，a group fed by starter diet 1 at d 7 and a group fed by starter diet 1 at d 42，and lambs from both groups were weaned at d 56 and transacted to starter diet 2 at d 60.The transition period lasted 10 d.Six lambs from each group were slaughtered at d 14，28，42，56，70 and 84，respectively.The ventral sac of the rumen was collected，rumen papilla height，width and muscular thickness were measured，and total RNA was extracted.Real-time qPCR was used to analyze the expression of genes related to rumen papilla growth.Tissue sections were prepared to observe rumen development.The results revealed that rumen papilla width of 7 d-starter group was higher (P<0.05) than that of 42 d-starter group，however，rumen muscular thickness in 7 d-starter group was lower (P<0.05) than that in 42 d-starter group.The IGFBP3 expression of rumen tissue in 7 d-starter group were lower (P<0.05) than that in 42 d-starter group.In contrast，IGFBP5 expression in 7 d-starter group were higher (P<0.05) than that in 42 d-starter group.In 7 d-starter group，rumen papilla height and width were positively correlated with expression of TGFβ1 and IGFBP3 (R=0.507，P<0.001；R=0.444，P<0.001；R=0.465，P<0.001；R=0.299，P=0.011)，while were negatively correlated with expression of IGFBP6 (R=－0.443，P<0.001；R=－0.477，P<0.001).In 42 d-starter group，rumen papilla height was positively correlated with expression of IGFBP5 (R=0.227，P=0.018)，additionally，rumen papilla width were positively correlated with expression of IGFBP3 and IGFBP5 (R=0.338，P=0.005；R=0.293，P=0.002).In conclusion，this study result indicate that starter feeding for lambs at day 7 can improve rumen morphology，and the expression levels of genes involved in cellular growth in ruminal epithelium can regulate rumen development.

To establish an method for rapid and accurate detection of the yak enterovirus, the yak enterovirus SWUN-AB001 obtained by our laboratory was used to immunize healthy rabbits, and then establish an indirect immunofluorescent assay (IFA) by using the rabbit hyperimmune serum. And the working conditions and time were screened and optimized. The best dilution and working time of hyperimmune serum in the IFA method were 1:100 and 30 minutes, and these of FITC conjugated goat anti-rabbit IgG were 1:800 and 45 minutes. Specificity of bright green fluorescent particles about the yak enterovirus were clearly visible; it can′t react with Bovine viral diarrhea virus, Rotavirus, E. coli, Salmonella, Streptococcus, Staphylococcus and Pasteurella; the minimum concentration of SWUN-AB001 can be detected by the method was about 20TCID₅₀•mL^-1. Forty clinical diarrhea samples were detected by using the RT-PCR and the IFA, and there was no difference in the experiment results between the RT-PCR and the IFA (P=0.329). The results indicate that the method has strong specificity, good repeatability and high sensitivity, it can be used in study of epidemiological investigation, clinical diagnosis and laid a foundation of the fast diagnostic reagents.

This experiment was developed to simultaneously detect these nine viral pathogens that cause infections in ducks including avian influenza virus (AIV) subtypes H5, H7, and H9; duck hepatitis A virus (DHAV-A); duck Tembusu virus (DTMUV); duck enteritis virus (DEV); egg drop syndrome virus (EDSV); Newcastle disease virus (NDV); duck circovirus (DuCV); muscovy duck reovirus (MDRV); and muscovy duck parvovirus (MDPV). Twelve pairs of specific primers were designed according to the conserved sequences of the genes from each pathogen available in the GenBank database. Single, mixed pathogen cDNA/DNA templates, other common duck pathogens and duck organs nucleic acids were used to evaluate the specificity of the GeXP-multiplex assay. Serial dilution from 10⁶ to 10¹ copies•μL^-1 of in vitro transcript RNA of target genes and plasmids were used to test the sensitivity of GeXP multiplex PCR assay. Different amounts of the templates (10³ and 10⁷ copies•μL^-1) were selected at random, mixed and tested in the GeXP multiplex PCR assay. The results were compared with those of the single template GeXP multiplex PCR assay. The GeXP assay was evaluated using 150 clinical specimens and compared with the single PCR. All positive specimens in the GeXP multiplex PCR and conventional PCR were identified via sequencing to verify the accuracy and reliability of the GeXP assay. These results showed that the corresponding specific fragments of genes were amplified by the single and multiplex GeXP PCR assay. Other pathogens did not result in amplification products. The detection limit of GeXP was 10³ copies•μL^-1 when all nine types of duck virus were present. The results of the interference assay showed that three specific amplification peaks can still be observed in the case of combination of three different concentration templates. The results of these experiments showed that mixed infections can be detected by GeXP multiplex PCR with minimal interference. Compared with the results of conventional PCR, the GeXP multiplex PCR method was more sensitive and accurate in the detection of 150 clinical samples. In conclusion, this GeXP-based multiplex PCR is a high-throughput, specific and sensitive test to detect nine duck viruses. This assay provides a new method in rapid molecular diagnosis for mix clinical duck virus samples.

Newcastle disease virus (NDV) is the pathogen of Newcastle disease (ND), which causes serious economic losses to poultry industry. For a better understanding of innate immune response in the pathogenesis of NDVs, SPF chicken embryos were infected with a virulent NDV strain and an avirulent vaccine strain-LaSota, respectively, with different infection doses set for both viruses. At the early stage of viral infection, the dynamics of viral proliferation and the transcriptional level of innate immune-related molecules were tested with fluorescence quantitative PCR assays. The results showed that both the virulent and avirulent NDV strains could activate the innate immune signaling pathways, and resulted in the transcriptional changes of cellular pattern recognition receptors, interferons, interleukins and antiviral proteins. NDV infection of high doses in comparison with low doses resulted in quicker viral proliferation and earlier peaks of innate immune-related gene expression. The virulent NDV strain induced higher levels of interferons and inflammatory cytokines in chicken embryos, which was consistent with the severe inflammation and pathological damages of both chicken embryos and chickens caused by virulent viral infection. In conclusion, this study found that the response patterns of chicken innate immune signaling pathways varied due to NDV virulence and infection dose, and the genotype VII NDV strain could induce strong cellular inflammatory responses, which may contribute to its high pathogenicity to chickens.

A multiple antigenic peptide (MAP), composed of six B-cell epitopes and three T-cell epitopes derived from antigenic proteins of Mycoplasma capricolum subsp. capripneumoniae (Mccp), was designed and expressed in Escherichia coli cells, which was then injected into mice to evaluate humoral and cellular immune responses induced by the recombinant MAP. Antibody levels against MAP, each single B-cell epitope, and the ultrasonic-treated crude Mccp antigen were all significantly increased (P<0.01). Results of in vitro metabolic inhibition test indicated that diluted sera (1:64) from immunized mice inhibited Mccp growth. Splenic lymphocytes from the immunized mice exhibited remarkable proliferation when stimulated by each single T-cell epitope and crude Mccp antigen (P<0.01). The production of IFN-γ, TNF-α, IL-1β, and IL-10 were increased (P<0.001), while that of IL-12 was decreased (P<0.001). These data showed that the constructed MAP stimulated immune responses in mice. The results laid a certain foundation to develop subunit vaccine against contagious caprine pleuropneumonia caused by Mccp.

The mRNA relative expression and tissue distribution of wnt gene family wnt1, wnt2, wnt4, wnt5 and wnt11B in different proglottides of Moniezia expansa were investigated in the present study to provide basic information for exploring the roles of the wnt signaling pathway in development of M. expansa. The relative mRNA expression of wnt1, wnt2, wnt4, wnt5 and wnt11B in different proglottides of M. expansa were determined by qRT-PCR. Tissue location of wnt1, wnt2, wnt4, wnt5 and wnt11B in different proglottides of M. expansa were detected using in situ hybridization. The quantitative results showed that the gene family of wnt were transcribed in all proglottides of adult M. expansa. The transcription of wnt1, wnt4, wnt5 and wnt11B were higher in scolex than other proglottides, and in the gravid proglottides, the transcription of these genes was lowest. On the contrary, the transcription of wnt2 was lowest in the scolex, but higher in other proglottides. The in situ hybridization results showed that the wnt gene family mRNA were mainly distributed in suckers of scolex; in the mature proglottides, they were mainly distributed in ovary, testis and interproglottidal; in the immature and gravid proglottides, they were mainly distributed in interproglottidal. Based on these findings, the mRNA relative expression of these 5 wnt genes in scolex and immature proglottides was higher than in other proglottides, and mainly distributed in suckers, interproglottidal and germ cells. The results suggested that the Wnt signaling pathway may be involved in development of segments and germ cell in M. expansa, which remains to be further verified.

The aim of the study was to observe comparatively the ultrastructural changes of hook and no-hook cysticercus pisiformis (C. pisiformis) under relative rest and moton. The well-developed C. pisiformis were obtained in the procedure of slaughter inspection and autopsy. The C. pisiformis was taken out by cutting cyst directly, or cultured with normal saline containing 10% rabbit’s bile until evagination. Then samples were made, observed and recorded by scanning electron microscopy. Ultrastructural changes of hook and no-hook C. pisiformis mainly appeared on rostellum. The head end of hook C. pisiformis was relatively dull, and connected antlers-liking hook through cutaneous muscular columns. In the mild movement the cutaneous muscular columns on rostellum contracted, hooks were outward stretch and the head end of rostellum become flat. When cutaneous muscular columns sostenuto contracted, the head end of rostellum began to invaginate, hook outspread toward diagonal upward side and formed bell mouth-like structure. When rostellum powerfully contracted, the cutaneous muscular column got into the rostellum and become a cylinder, hook arranged along cylinder mouth and formed chrysanthemum-like structure. At relative rest the restellum of no-hook C. pisiformis was flat, liking several layers of oblaten stacked together, there was a round nodule in central part and gear-like decorative design on periphery. When rostellum mild exercised the nodule in central part invaginated and presented valvula-like structure, periphery of rostellum had half round cyclic structure and looked like a life buoy. When rostellum was in strong movement, the valvula-like structure of nodule was opened and looked like a bell mouth. Tegument around rostellum expanded toward outside, looking like a tyre structure. There were several radial folds between the bell mouth of rostellum and tyre-like structure. The rostellum turned into wheel-like structure. The suckers of hook and no-hook C. pisiformis were located behind restellum and had three contraction patterns that were annular contraction, longitudinal contraction and radical contraction. Ultrastructural structure of reostellum of hook and no-hook C. pisiformis is totally different and respective special structural changes appear in the different motion state.

Cytochrome P450 is a superfamily of enzymes responsible for the metabolism of drugs, xenobiotics and endogenous compounds. In them, the member of the CYP2D6 constitutes only about 2% of total hepatic CYPs, however, it is responsible for the metabolism of about 20% of commonly prescribed therapeutic compounds. The purpose of this research was to systematically study the activity of bactrian camel CYP2D6 enzyme and its sensitivity to the specific inhibitors in vitro. Liver microsomes of bactrian camels was prepared by modified differential centrifugation method and the protein concentration, total content of CYP enzyme were measured by BCA method and CO reduction method, respectively. Then the characteristics of Bactrian camel CYP2D6 enzyme kinetics, substrate metabolic rate and inhibitory effect of quinidine on the enzyme were studied respectively on the basis of optimization of liver microsomal system and the establishment of the HPLC method for testing the specific substrate dextromethorphan hydrobromide of CYP2D6 enzyme and its active metabolite dextrophan. The results showed that the both liver microsomal protein concentration and the total CYP enzyme content could meet the study requirements. The established HPLC method for determining the specific substrate of CYP2D6 enzyme and its metabolite was high sensitivity and stability, as well as all the peaks of each component were separated completely with no interference. The V_max value and K_m value of CYP2D6 enzyme were 0.141 6±0.052 7 nmol•(min•mg)^－1 and 12.986 0±0.357 2 μmol•L^－1, respectively. The IC₅₀ value of quinidine was 2.779 0±0.063 9 μmol•L^－1. Therefore, the incubation system of Bactrian camel liver microsomal was established and optimized successfully for the further studying of the CYP2D6 enzyme activities in vitro, and found out the kinetic parameters of Bactrian camel CYP2D6 enzyme and the specific inhibitor's IC₅₀ value of the enzyme. Hence, the experimental results filled the blank in research area of CYP2D6 enzyme activities in vitro.

The Real-time PCR and Immunohistochemistry were used to examine the expression and localization of Ghrelin in the reindeer organs. The results of Real-time PCR indicated that Ghrelin was expressed in 17 kinds of organs and the expression in the abomasum was highest, followed by pancreas, duodenal, testis and oesophagus, in which the expression of Ghrelin were remarkably higher than that of the rest organs (P<0.05). Immunohistochemical results showed that Ghrelin-positive cells in the oesophagus and abomasum mainly distributed in the tunica mucosa, tunica submucosa and tunica muscularis and also detected in the tunica mucosa, tunica submucosa of rumen, reticulum, omasum. In the intestine, Ghrelin-positive cells were mainly localized in the tunica mucosa, tunica submucosa and tunica muscularis of duodenum, jejumum, ileum, colon, especially more cells were found in the epithelium of intestinal villi and the tunica submucosa. Ghrelin-positive cells were observed in the liver, pancreas, lung, spleen, kidney, testis, anterior pituitary, thyroid gland. Real-time PCR and immunohistochemistry results showed that Ghrelin was presented in all of the organs that were examined, where the expression and distribution of Ghrelin in the esophagus and gastrointestinal were most widely, suggesting its possible role in regulating the digestive function.

Two cases of pet dogs were confirmed to be infected with Brucella canis based on detection to their excretion by use of the BRU real time PCR and conventional PCR kits, and the partial genetic sequences of the Brucella canis infected were characterized. The results showed that the Brucella strains from the pet dogs belong to the Brucella canis, and atypical canine brucellosis. Phylogenetic analysis of BSCP31 gene sequences showed 99%-100% nucleotide identity with the published sequences in GenBank, and OMP25 gene showed 99.9% nucleotide identity between Brucella canis xue1 strain and Brucella canis HSK A52141 strain. Sequence analysis on 16S rRNA showed the strains from French Bulldog and Teddy Dog belong to Brucella. These data might provide a valuble infomation for future screening and prevention of canine brucellosis.