Abstract:The objective of this study was to evaluate the effect of diet supplementation with potassium nitrate on rumen gas production in vitro and bacterial diversity in vivo in in sheep. First, rumen fluid was obtained from 3 cannulated Hu sheep fed basal diet to compare gas production of in vitro diet contained nitrate 0%(A, control group), 1% (B), 2% (C), 3% (D), 4% (E), 5% (F), respectively. In a second experiment, 3 cannulated sheep were fed the six diets above, with nitrate concentration increased gradually. Each period had 16 days; before morning feeding at the end of each period, rumen fluid was collected for DNA extraction, 16S rRNA Illumina sequencing and qPCR analysis. In vitro study showed that there was no significant difference in the net gas production, theoretical maximum gas volume and speed of gas production among groups, however, the net gas production and theoretical maximum gas volume of 5% group tended to declined(P<0.1). Paired-End sequencing of 16S rRNA gene targeting showed that Chao1 index of 5% group was significantly higher than control. No difference in Shannon index was observed. At the phylum level, the total percentage of Bacteroidetes, Firmicutes, Proteobacteria, Fibrobacteres, Verrucomicrobia was more than 90%. At the genus level, Prevotella, Selenomonas, Succinivibrio were dominant genus in each group, the proportion was 47.87%, 5.76%, 2.37% respectively. qPCR showed increase in the relative abundance of Campylobacter fetus with nitrate concentration increased (P<0.01). These results indicate that nitrate addition ranged from 0% to 5% change the abundance of bacteria, especially nitrate reduction bacteria, but do not change gas production, rumen bacterial diversity and advantages community structure.

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	1. Effect of dietary nitrate dosage on gas production in vitro and rumen bacterial abundance of Hu Sheep
	Chen Zhiyuan,Ma Tingting,Gong Xiaoxiao,Su Xiaoshuang,Li Guodong,Zhao Guoqi,Lin Miao
	Animal Husbandry, Veterinary Medicine 24 August 2015
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	Abstract:The objective of this study was to evaluate the effect of diet supplementation with potassium nitrate on rumen gas production in vitro and bacterial diversity in vivo in in sheep. First, rumen fluid was obtained from 3 cannulated Hu sheep fed basal diet to compare gas production of in vitro diet contained nitrate 0%(A, control group), 1% (B), 2% (C), 3% (D), 4% (E), 5% (F), respectively. In a second experiment, 3 cannulated sheep were fed the six diets above, with nitrate concentration increased gradually. Each period had 16 days; before morning feeding at the end of each period, rumen fluid was collected for DNA extraction, 16S rRNA Illumina sequencing and qPCR analysis. In vitro study showed that there was no significant difference in the net gas production, theoretical maximum gas volume and speed of gas production among groups, however, the net gas production and theoretical maximum gas volume of 5% group tended to declined(P<0.1). Paired-End sequencing of 16S rRNA gene targeting showed that Chao1 index of 5% group was significantly higher than control. No difference in Shannon index was observed. At the phylum level, the total percentage of Bacteroidetes, Firmicutes, Proteobacteria, Fibrobacteres, Verrucomicrobia was more than 90%. At the genus level, Prevotella, Selenomonas, Succinivibrio were dominant genus in each group, the proportion was 47.87%, 5.76%, 2.37% respectively. qPCR showed increase in the relative abundance of Campylobacter fetus with nitrate concentration increased (P<0.01). These results indicate that nitrate addition ranged from 0% to 5% change the abundance of bacteria, especially nitrate reduction bacteria, but do not change gas production, rumen bacterial diversity and advantages community structure.
	TO cite this article:Chen Zhiyuan,Ma Tingting,Gong Xiaoxiao, et al. Effect of dietary nitrate dosage on gas production in vitro and rumen bacterial abundance of Hu Sheep[OL].[24 August 2015] http://en.paper.edu.cn/en_releasepaper/content/4652935


	2. Insulin stimulates goose liver cell growth by activating PI3K-AKT-mTOR signal pathway
	HAN Chunchun,LIU Dandan,HE Fang,WAN Huofu
	Animal Husbandry, Veterinary Medicine 05 December 2014
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	Abstract:Our previous result showed in the fatty liver formation induced-by overfeeding goose, it was accompanied by an activation of PI3K-Akt-mTOR pathway and an increase of plasma insulin concentration. Recent studies have suggested a crucial role for PI3K-Akt-mTOR pathway in regulating cell proliferation, so we hypothesize that insulin acts goose hepatocellular growth by PI3K-Akt-mTOR signal pathway. Here we first showed that insulin evidently increased the cell DNA synthesis rate, rate of BrdU-positive cells, the mRNA level and protein content of factors involved in the cell proliferation of goose primary hepatocytes. Meanwhile, insulin evidently increased the mRNA level and protein content of factors involved in PI3K-Akt-mTOR pathway. However, the up-regulation of insulin on cell proliferation was decreased significantly by the inhibitors of PI3K-Akt-mTOR pathway, LY294002, rapamycin or NVP-BEZ235. These findings suggest that PI3K-Akt-mTOR pathway plays an essential role in insulin-regulated cell proliferation of goose hepatocyte.
	TO cite this article:HAN Chunchun,LIU Dandan,HE Fang, et al. Insulin stimulates goose liver cell growth by activating PI3K-AKT-mTOR signal pathway[OL].[ 5 December 2014] http://en.paper.edu.cn/en_releasepaper/content/4619499

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	3. Expression and methylation status of imprinted gene IGF2R in lung of deceased cloned transgenic dairy goat
	ZHANG Yanli,ZHANG Guomin,WAN Yongjie,Wang Feng
	Animal Husbandry, Veterinary Medicine 20 November 2014
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	Abstract:Dairy goat is a good model for production of transgenic proteins in milk by somatic cell nuclear transfer (SCNT). However, animals produced from SCNT are often linked with lung deficiencies. Imprinted genes are important regulators of lung growth, and may be subjected to faulty reprogramming. In the present study, firstly, microsatellite analysis, PCR and DNA sequence identification were conducted to confirm that these three dead kids were genetically identical to the transgenic donor cells. Secondly, the CpG island methylation profiles of imprinted insulin-like growth factor receptor (IGF2R) gene was assessed by Bisulfite Sequencing PCR (BSP) in lungs of the three dead transgenic clones and normally produced lambs. In addition, the relative mRNA level of IGF2R was also determined by real-time PCR (RT-PCR). Results showed that IGF2R gene in the lung of dead cloned goat fetus showed abnormal hypermethylation and higher mRNA expression levels (P<0.05) comparing to the control, indicating that aberrant DNA methylation reprogramming may be one of the important factors for the death of transgenic cloned animals.
	TO cite this article:ZHANG Yanli,ZHANG Guomin,WAN Yongjie, et al. Expression and methylation status of imprinted gene IGF2R in lung of deceased cloned transgenic dairy goat[OL].[20 November 2014] http://en.paper.edu.cn/en_releasepaper/content/4619349


	4. Effects of Cold Stress on the Intestinal Microbiota and Morphology in Quail Cecum
	FuJing,Liu Chunpeng,XuShiwen
	Animal Husbandry, Veterinary Medicine 22 September 2014
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	Abstract:The objective of the current study was to determine the influence of cold stress on cecal microbial populations and morphology in quails. The normal microflora of intestinal is considered very important for animal health and growth performance. In modern poultry production systems, environmental stressors may influence bird performance and susceptibility to pathogens. A total of 192 quail with 15 days of age were randomly allocated into 12groups (16 each group) for exposure to acute (up to 12 h) and chronic (up to 20 days) cold temperature (12±1 C). Changes in commensal intestinal microbial populations were determined by denaturing gradient gel electrophoresis assay, and alterations in cecum morphology were observed in histopathology. The data showed that under acute cold stress, although the microbial community structure is altered, but the change was not significant. However, chronic cold stress induced the microbial community structure from simple to complex effectively. Morphologically, significant damage was also observed in cecum after both acute and chronic cold stresses to the quails. The data from the current study indicated that both acute and chronic cold stress were effect on the cecal microbiota which might be due to the cold-damaged intestinal morphology.
	TO cite this article:FuJing,Liu Chunpeng,XuShiwen. Effects of Cold Stress on the Intestinal Microbiota and Morphology in Quail Cecum[OL].[22 September 2014] http://en.paper.edu.cn/en_releasepaper/content/4609784


	5. Expression of Nerve Growth Factor (NGF), and Its Receptors TrkA and p75 in the Reproductive Organs of Laying Hens
	PU Shaoxia,QU Changwei,LI Chunmei
	Animal Husbandry, Veterinary Medicine 10 July 2014
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	Abstract:In order to investigate the expression levels of nerve growth factor (NGF) and its receptors (TrkA and p75) in the prehierarchical follicle and oviducts of hens, five 130-day-old laying hens were examined by immunohistochemistry and RT-PCR analyses. NGF and its receptors were expressed in the theca cells and granulosa cells of prehierarchical follicles, and they also expressed in the epithelial cells of oviducts. The expressions of NGF mRNA were discrepant in different parts of the prehierarchical follicle and oviducts. The expression of NGF and p75 mRNA levels in large white follicle (LWF) were high, as well as TrkA in small yellow follicle (SYF). In oviduct, the expression of NGF is highest in infundibulum but lowest in magnum. These results suggest that NGF may play an important role in the regulation of reproduction in hens.
	TO cite this article:PU Shaoxia,QU Changwei,LI Chunmei. Expression of Nerve Growth Factor (NGF), and Its Receptors TrkA and p75 in the Reproductive Organs of Laying Hens[OL].[10 July 2014] http://en.paper.edu.cn/en_releasepaper/content/4603982


	6. In vitro culture and induced differentiation of goat fetal lung-derived mesenchymal stem cells (LMSC)
	Gao Jing,Liu Pengxia,Tai Dapeng,Cang Ming,Liu Dongjun
	Animal Husbandry, Veterinary Medicine 24 February 2014
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	Abstract:Mesenchymal stem cells (MSCs) occur in a variety of mammalian fetal and adult tissues. To date, most studies have been carried out on bone marrow-derived MSCs, with few studies reported on fetal lung-derived MSCs (LMSC). The purpose of this study was to separate fibroblastic cells from goat fetal lung tissue; culture these cells in vitro; identify their MSC characteristics; induce differentiation of goat fetal LMSCs into osteoblasts, adipocytes, chondrocytes, islet β-cells, and neurons; and validate their pluripotency. Our findings confirmed that the separated goat fetal LMSCs were fibroblasts capable of efficient growth and proliferation. RT-PCR showed positive expression of the following MSC marker genes compared to the control group: CD13, CD29, CD44, CD90, and CD106, and negative expression of CD45. Induced differentiation of goat fetal LMSCs into adipocytes, osteoblasts, chondrocytes, islet β-cells, and neurons was determined by cytochemical staining methods using Oil Red-O, silver nitrate (AgNO3), alcian blue, dithizone, and neuron-specific enolase (NSE), respectively; and confirmed by RT-PCR identification of the marker genes: PPARγ2, collagen I, COL2A1, PDX1, and neuron-specific NSE, respectively. In conclusion, this study showed that goat fetal lung-derived cells possessed the biological characteristics and multipotent differentiation potential of MSCs. In addition, we have established successful methods for the separation, purification, and validation of goat fetal LMSCs, thereby preparing the ground for further cell cloning therapies and associated animal model studies.
	TO cite this article:Gao Jing,Liu Pengxia,Tai Dapeng, et al. In vitro culture and induced differentiation of goat fetal lung-derived mesenchymal stem cells (LMSC)[OL].[24 February 2014] http://en.paper.edu.cn/en_releasepaper/content/4587206


	7. Evaluation of reference genes for investigation of gene expression profiles in chorioallantois from Meishan and Yorkshire pigs on gestational days 26 and 50
	HOU Chunyan,YU Mei,LI Xiaoping
	Animal Husbandry, Veterinary Medicine 24 December 2013
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	Abstract:Selection of suitable reference gene is important for accurate measurement of gene expression data obtained using the quantitative real-time PCR (qPCR) method. There has been no systematic investigation of reference genes for porcine chorioallantois which plays important roles in mediating concepts/fetus growth and survival. In this study, six candidate reference genes expressed in pig chorioallantois from Meishan and Yorkshire pigs on gestational days 26 (GD26) and 50 (GD50) were evaluated by qPCR. These genes included TATA box binding protein (TBP), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), ribosomal protein L32 (RPL32), actin-beta (ACTB), ribosomal protein S20 (RPS20) and RNApolymerase polypeptide G (RPG). The expression stability of these genes was investigated by using the GeNorm software program and statistical analyses. The rank order of the expression stability for these genes was (from the most stable to the least stable): TBP, GAPDH, RPL32, ΒACTIN, RPS20 and RPG. Collectively, the results suggest that the TBP and GAPDH could be considered as the suitable reference genes for investigation of gene expression in the pig chorioallantois from the two breeds at different gestational stages.
	TO cite this article:HOU Chunyan,YU Mei,LI Xiaoping. Evaluation of reference genes for investigation of gene expression profiles in chorioallantois from Meishan and Yorkshire pigs on gestational days 26 and 50[OL].[24 December 2013] http://en.paper.edu.cn/en_releasepaper/content/4577871


	8. Effects of Ultradry Storage on Fluidity of Plasma Membrane of Haloxylon persicum Seeds
	TONG Lirong
	Animal Husbandry, Veterinary Medicine 08 August 2013
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	Abstract:The DPH fluorescent probe (1,6-diphenyi-1,3,5,-hexatriene) was used to study the the effects of ultradry seed storage on the fluidity of plasma membrane. Results indicated that the micro-viscosity of plasma membrane of ultradried seeds had no significant changes compared with the Haloxylon persicum seeds which were stored under 4℃ condition. However, there was a little adverse effect on the seeds with extreme dehydration. The results were consistent with higher vigor level of ultradried seed. It indicated that ultradry seed storage could maintain the physiological function of seed, protect the integrity of the membrane and improve the storability of seed.
	TO cite this article:TONG Lirong. Effects of Ultradry Storage on Fluidity of Plasma Membrane of Haloxylon persicum Seeds[OL].[ 8 August 2013] http://en.paper.edu.cn/en_releasepaper/content/4554086


	9. The total AvBD-6 gene expressed by Pichia pastoris
	Wenyu Sun,Anshan Shan,Qingquan Ma,Na Dong,Yanping Cao,Liang Wang,Zeyun Wang,Changjiang Yan
	Animal Husbandry, Veterinary Medicine 09 January 2013
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	Abstract:Avian β-defenisin 6 (AvBD-6) is a 67 amino acid-long peptide that plays an important role in the innate immunity of chickens and is widely expressed in most tissues. In this study, the codon-optimized AvBD-6 gene was synthesized and cloned into the eukaryotic expression vector pPICZαA and expressed in Pichia pastoris. Expression levels of the recombinant AvBD-6 were tested by Tricine-SDS-PAGE, and the product concentration reached 213.0 mg/l after optimization of the expression conditions. Recombinant AvBD-6 exhibited antimicrobial activity against bacteria with the minimal inhibition concentration (MIC) from 5.8 to 46.6 μg/ml and showed no hemolytic activity.
	TO cite this article:Wenyu Sun,Anshan Shan,Qingquan Ma, et al. The total AvBD-6 gene expressed by Pichia pastoris[OL].[ 9 January 2013] http://en.paper.edu.cn/en_releasepaper/content/4512514


	10. Studies on Physiological Characteristics of Aging-resistence of Ultradried Ceratoides arborescens Seeds
	TONG Lirong,DONG Kuanhu
	Animal Husbandry, Veterinary Medicine 14 July 2012
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	Abstract:Ceratoides arborescens seeds were tested for the effects of aging-resistance and free radical scavenging following seed ultradry. The results indicated that: (1)Ultradried seeds under the conditions of certain moisture content showed no adverse effect on seed vigor. Furthermore, ultradried seeds significantly enhanced aging-resistant capability under aging-accelerating condition (45℃, 2 days). The critical moisture content for Ceratoides arborescens seeds was about 3.78%. (2) Activities of the antioxidant enzymes including catalase and superoxide dismutase were reserved during storage in an ultradry state. (3) During ultradrying of Ceratoides arborescens seeds, the level of free radicals increased, but decreased as prehumidification before imbibition, which had a scavenging action on free radicals.
	TO cite this article:TONG Lirong,DONG Kuanhu. Studies on Physiological Characteristics of Aging-resistence of Ultradried Ceratoides arborescens Seeds[OL].[14 July 2012] http://en.paper.edu.cn/en_releasepaper/content/4484927

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