Acta Veterinaria et Zootechnica Sinica

Genetic Identification of Swamp-River Buffalo Hybrids Based on Microsatellite DNA Markers

ZHANG Yi;SUN Dong-xiao;YU Ying;ZHANG Yuan

2009, 40(4): 453-458. doi:

Abstract ( 814 )

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In order to develop a molecular method to detect swamp-river buffalo hybrids using genetic markers and characterize the status of Chinese buffalo recourses, thirty microsatellite markers were genotyped in 992 individuals that represented 18 indigenous swamp buffalo populations, 2 exotic river buffalo breeds, and 1 crossbreeding population. Results reflected that there existed considerable genetic differentiation between two buffalo types with F_ST = 0.310±0.193. Based on microsatellite multi-locus genotypes, both the model-based Bayesian STRUCTURE analysis and the multivariate correspondence analysis could sensitively distinguish pure swamp or river buffalo and hybrid individuals. Moreover, results of genetic admixture analysis indicated that all Chinese local buffalo populations generally showed pure swamp ancestry. These findings may provide a technique basis for objectively assessing Chinese swamp buffalo genetic resources and making conservation decisions.

Evaluation of Selection Schemes for Beef Line of Luxi Cattle

ZHANG Qing-feng;XU Shang-zhong

2009, 40(4): 459-467. doi:

Abstract ( 741 )

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On the basis of the current two-tire nucleus breeding scheme and young bull system for beef line of Luxi cattle, the backfat thickness and eye muscle area of rib scanned in yearling live cattle and mean concentration of IGF-Ⅰin plasma at 28th,42nd and 56th day of post-weaning period in calf were used as early records of carcass traits in this study. Four selection schemes which differed in the records available for use as selection criteria were defined, and deterministic simulation was used to evaluate 4 selection schemes for breeding efficiency. The results showed that taking if the scheme 1 (current scheme) contrast with other schemes, then index accuracy, genetic gains per year and profitability were improved 0.19, 39.05％ and 63.60％ for scheme 2; 0.01, 25.55％ and 14.90％ for scheme 3; 0.21, 40.68％ and 64.08％ for scheme 4. According to the results, the schemes taking the ultrasound scanning and IGF-Ⅰas selection criteria would get more ideal breeding efficiency in the breeding schemes.

Bone Morphogenetic Protein 15 as a Candidate Gene for Prolificacy of Jining Grey Goat

FENG Tao;ZHAO You-zhang;CHU Ming-xing;DI Ran;ZHANG Ying-jie;FANG Li

2009, 40(4): 468-475. doi:

Abstract ( 1086 )

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According to the sequence of sheep bone morphogenetic protein 15 (BMP15) gene, two pairs of primers (P1 and P2) were designed to amplify the exons 1 and 2 of BMP15 gene in five randomly selected individuals from Angora and Jining Grey goats, respectively. Sequencing showed that sequences of BMP15 exon 1 (P1 amplification) of Angora and Jining Grey goats were identical. There were three nucleotides (CTT) insertion from 268 to 270 positions in goat BMP15 exon 1 compared with that of sheep (AF236078), which caused a leucine insertion at the 12th position in amino acid sequence. Sequence length of goat BMP15 exon 2 (P2 amplification) was identical with that of sheep (AF236079), but there were seven nucleotide and four amino acid changes between goat and sheep. The nucleotide at the 963rd position in BMP15 exon 2 was A for Angora goat and sheep, and G for Jining Grey goat. Based on A963G mutation,primer P3 was designed to detect SNP of BMP15 exon 2 in high prolificacy breed (Jining Grey goat), moderate prolificacy breed (Boer goat) and low prolificacy breeds (Angora and Inner Mongolia Cashmere goats) by PCR-SSCP. Three genotypes (AA, AG and GG) were detected in Jining Grey goats, two genotypes (AG and GG) in Boer goats, only AA genotype in Angora and Inner Mongolia Cashmere goats. Sequence analysis revealed one mutation (A963G) in genotype GG compared with genotype AA, and this mutation resulted in an amino acid change of serine→glycine (S300G). In Jining Grey goats, frequencies of AA, AG and GG genotypes were 0.008, 0.059 and 0.933, respectively. Genotype distributions of BMP15 gene were significantly different (P<0.05 or P<0.001) among Jining Grey goats and Boer, Angora, Inner Mongolia Cashmere goats. In Jining Grey goats, the individuals with genotype GG had 0.71 (P<0.05) or 1.57 (P<0.05) kids more than those with genotypes AG or AA, the individuals with genotype AG had 0.86 (P<0.05) kids more than those with genotype AA. These results preliminarily indicated that the BMP15 gene is either a major gene which affects the prolificacy in Jining Grey goats or a molecular marker in close linkage with such a gene.

Polymorphism of the Prolactin Receptor (PRLR) Gene and Its Association with Litter Size in Swine

CAO Guo-qing;LI Bu-gao;SHI Jian-zhong;LIU Hong;LIU Jian-hua;ZHOU Zhong-xiao

2009, 40(4): 476-480. doi:

Abstract ( 1462 )

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The aim of this study was to detect the polymorphism in exon 10 of porcine PRLR gene and study the effects of the polymorphic site on litter size traits in swine. A total of 221 pigs from Large White, Mashen, Shanxi Black and Shanxi White pig populations were used to detect the polymorphism of the exon 10 of PRLR gene by PCR-RFLP, and the simple univariate animal model was used to analyze the effects of the PRLR genotypes on total number born (TNB) and number born alive (NBA) at first parity. The results showed that there was a NaeⅠ polymorphic site within the exon 10 of PRLR gene. Two alleles of A and B, and two genotypes of AA and AB, were detected in examined populations. The A allele was the superior allele in any one of the four populations, with the frequencies ranging from 0.86 to 0.93. The PRLR genotype was significantly associated with TNB and NBA of first parity. Sows with AA genotype had 1.797 and 1.293 more TNB and NBA than those of sows with AB genotype (P<0.05). This implied that the reproduction performance of pig could be improved if the NaeⅠ polymorphic site detected in this study was applied as a genetic marker in pig breeding and production.

An Alu Insertion Polymorphism in the Porcine FSHβ and GDF9 Gene

ZHANG Lu;CAO Hong-zhan;LU Chun-lian;HU Cheng-bo

2009, 40(4): 481-486. doi:

Abstract ( 741 )

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The Alu insertion sequence of the porcine FSHβ and GDF9 genes were obtained using the PCR(Polymerase chain reaction) technique, from Liaoning Black，Landrace，Large White，Danyu Black and Duroc pigs, and the polymorphism was investigated，and the sequence was analyzed by bioinformatics in this study. The GLM model was constructed to study the relationship between litter size and the polymorphism of the Alu. The result showed that litter size of individuals with dominant genotype BBDD were 15 more than that with genotype AACC（P<0.01）. Otherwise, the FSHβ and GDF9 genes were correlated deeply to the major gene influencing on the litter size, which may be considered as a genetic marker associating with pig litter size. The dominant genotype (BBDD) can improve the litter size traits.

The Genetic Distribution and Population Genetic Structure Analysis of Antiviral Resistant Mx Gene Locus in Twelve Chinese Indigenous Chicken Breeds

LI Hui-fang;CHEN Kuan-wei;HAN Wei;ZHU Yun-fen;ZHANG Xue-yu;WANG Qiang

2009, 40(4): 487-492. doi:

Abstract ( 685 )

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In the experiment,PCR-RFLP technique was used to detect the diffrences of the resistant and sensitive alleles distribution of S631N locus of Mx gene in twelve Chinese indigenous chicken populations.The results showed that the PCR-RFLP technique properly detected the mutation between resistant allele A and sensitive allele G.Among the twelve populations,the average frequencies of resistant allele A and sensitive allele G were 0.304 and 0.696,respectively.The average value of observed heterozygosis and Shannon′s index at this locus were 0.657 2 and 0.524 0,respectively.Except for Xianju chicken,the other eleven populations were all in Hardy-Weinberg equilibrium at this locus (P>0.05).Except for Baier chicken,the EwensWatterson test indicated that this locus was neutral in all the other populations.The dendrograms based on allele frequency divided the twelve indigenous chicken populations into three clusters,which reflected the difference and advantage of Mx antiviral property in twelve indigenous chicken populations.

Cloning，Expression and Bioinformatics Analysis of KIT Gene in Alpacas (Lama pacos)

ZHANG Qiao-ling;JIANG Jun-bing;FAN Rui-wen;DONG Chang-sheng

2009, 40(4): 493-499. doi:

Abstract ( 758 )

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Three primers were designed to amplify the fragments of exon10 to exon14, exon13 to exon17 and exon16 to exon19 in alpacas. These fragments were 414, 539 and 426 bp, respectively. 1 044 bp cDNA sequence of KIT gene, spanned from exon10 to exon19, was obtained by contig software, and submitted to GenBank (DQ450844). The nucleotide sequence was aligned with the corresponding part of other animals, and the similarity was above 94%, which indicated that the sequences were homologous. It was confirmed that the sequence obtained in this study was the KIT gene of alpacas. And the deduced polypeptide was got by the translation software and it coded 347 amino acids. The multiple sequence alignments revealed that the similarity of alpacas KIT sequence were above 95% with other species. Three amino acid substitutions were detected in the polypeptide coded by exon1019 of KIT gene in alpacas, which were I→V at the position of 2, S→P at the position of 201 and K→R at the position of 344. And bioinformatics analysis indicated that there was much alpha-helix in the secondary structure; And the protein encoded the mast/stem cell growth factor receptor. These results would be the foundation of further researching the association between KIT gene and coat color of alpacas.

Effects of Nitric Oxide on the Growth and Development of Porcine Preantral Follicles Cultured in Vitro

YAO Gui-dong;TAO Yong;ZHANG Xiao-rong;QUAN Qing

2009, 40(4): 500-506. doi:

Abstract ( 779 )

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In order to understand the effects of nitric oxide (NO) on the growth and development of porcine preantral follicle, we treated the follicles with different concentrations of sodium nitroprusside (SNP, 0, 0.001, 0.01, 0.1 and 1 mmol·L^-1), a nitric oxide donor. The results showed that the follicle diameter increased during in vitro culture, but there were no significant differences (P>0.05); the survival rate in 1 mmol·L^-1 SNP group was significantly lower than that in 1 μmol·L^-1 SNP group (61.61% vs 81.52%, P<0.05), but no significant differences were found between other treatments (P>0.05); the antrum formation rate in 1 μmol·L^-1 SNP group reached highest to 50% on day 4, and the rate in 1 μmol·L^-1 SNP group on day 6 was higher than that of 0.01 mmol·L^-1 SNP; In addition, the antrum formation rate in 1 μmol·L^-1 SNP group was significantly higher than that in 0.1 and 1 mmol·L^-1 SNP group (73.07% vs 50%, 47.62%, P<0.05). After 6 days′culutre, the oocyte normal rate in 1 mmol·L^-1 SNP group was significantly lower than that in 1 μmol·L^-1 SNP group (71.21% vs 48.18％，P<0.05), and there were no significant difference between other treatments (P>0.05). The recovery rate of cumulus cells oocyte complexes (COCs) in 1 μmol·L^-1 SNP group was significantly higher than that of control and any other treatments (37.27% vs 22.88%, 25.59%, 20.74% and 19.39%, P<0.05). The results indicated that during the in vitro culture of porcine preantral follicle, low concentration of NO which is released from SNP improves the oocyte growth, development and follicle antrum formation while high level of NO is toxic to follicle survival.

Effects of Glycerol Supplementation on Feed Intake, Lactation Performance and Metabolites in Early Lactation Holstein Dairy Cows

WANG Cong;HUANG Ying-xiang;LIU Qiang;HUO Wen-jie;YANG Xiao-min;HE Dong-chang;ZHANG Shuan-lin;ZHANG Yan-li

2009, 40(4): 507-514. doi:

Abstract ( 768 )

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Thirty-six multiparous Holstein dairy cows, blocked by lactation number, corrected milk yield in last lactation, and expected calving date, were designated into four treatments in a randomized block experiment. Graded levels (0,100,200, and 300 g·d^-1) of glycerol were supplemented in the four treatments. The effects of graded glycerol supplementation on feed intake, lactation performance, changes of body weight, energy balance, concentration of blood metabolites and urine ketones in early lactation Holstein dairy cows were observed. The results showed that average feed intake, milk yield, milk protein percentage, milk lactose percentage and milk dry matter percentage were not affected significantly by glycerol supplementation (P>0.05). Feed efficiencies of the 200 and 300 g·d^-1 glycerol supplementation treatments were higher than those of the 100 g·d^-1 treatment and the control (P<0.05). Milk fat percentage was significantly depressed in the 300 g·d^-1 treatment compared with those in the control and 100 g·d^-1 treatment(P<0.05). Cows receiving 200 and 300 g·d^-1 glycerol had a higher plasma glucose and lower plasma NEFA than those of the contol (P<0.05), and 300 g·d^-1 glycerol supplementation treatment had lower content of β-hydroxybutyrate than that of control(P<0.05). 200 and 300 g·d^-1 glycerol supplementation decreased the contents of ketones in excreted urine compared with the control (P<0.05). The energy balance was improved in cows receiving the 200 and 300 g·d-1 glycerol supplementation treatments as reflected energy balance calculations, and a lower body weight loss compared with cows receiving the control (P<0.05). Results of the present study indicated that glycerol supplementation was beneficial to early lactating cows in improving their energy balance status.

Developing Rule of Digestive Organs in Weaned Rex Rabbit from American Offspring

LI Qing-hong;REN Ke-liang;WANG Bao-jun;YE Zhi-yuan;ZHANG Ya-qiang;HAN Jun-wen

2009, 40(4): 515-521. doi:

Abstract ( 692 )

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An experiment involving a total of 250 weaned Rex Rabbits was conducted to investigate the development rule of digestive organs. The weight of digestive organs and villous height（VH）,crypt depth（CD）,intestinal wall thickness（IWT）and pepsin, amylase and trypsin activities in small intestine were measured weekly from 28 day-old. The model parameters of Gompertz and Logistic were fitted with software of CurveExpert 1.3. The results showed that：①The inflexion point of weight gain of digestive organs was from 30 to 60 d, and the duration of the fastest weight gain of small intestine, large intestine, liver and stomach occurred from 35 to 63 d，35 to 63 d, 63 to 70 d,49 to 56 d, respectively; and ultimate weight 20.619 7 g for stomach was gained on 98 d, and 71.958 2 g for liver on 122 d, 42.500 3 g for small intestine on 119 d, 46.905 7 g for large intestine on 120 d. Maturing order of digestive organs is small intestine, stomach, large intestine and liver. ②IWT of small intestine，VH of front and middle duodenum, jejunum and ileum，and CD of the front and middle duodenum and jejunum were affected significantly by days（P<0.01,or P<0.05）. ③Days had no significant influence on the activities of of pepsin trypsins and the starch enzyme in small intestine（P>0.05），the enzyme activity showed a increasing tendency with growth and kept steady-going on 60 d. The determing values of trypsin and amylase activities betwen different small intestinal sections were different, the highest for the jejunum, the second for ileum, the smallest for the duodenum. It was concluded that Gompertz model is better to describe development of the digestive organs in Rex Rabbits，the growth of Rex Rabbit digestive organs is closely related to days, and its inflexion point is on 60 d, the mature of the digestive organs is on 120 d, and the recovery of morphologic changes caused by weaning needs about 14 d.

Effect of Aluminum Salt on Ammonia Release from Poultry Litter

JIAO Hong-chao;ZHANG Hong-fang;LIN Hai

2009, 40(4): 522-527. doi:

Abstract ( 733 )

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To investigate the effect of aluminum salt on ammonia production from poultry litter in commercial broiler houses, aluminum was added into hen dung at three different levels: 0, 2% and 4% (aluminum (g)/dung(g)). After aluminum salt treatment, the pH value, ammonia production, urease activity of treated dung and the content of ammonia nitrogen or total nitrogen in treated dung were measured respectively at indicated days. We found that ammonia released from dung and the pH value of dung decreased significantly after treatment compared with that of control dung. Furthermore, the urease activity in dung was significantly inhibited with a time course dependent manner after aluminum salt treatment (at 7 and 14 d ). In addition, the total nitrogen content, but not ammonia nitrogen content in dung was significantly decreased (P＜0.05) after 3 days′ treatment, especially in high level (4%) treatment group. These results suggested that pH value reduction, urease activity suppression and total nitrogen consumption induced by aluminum addition contributed to the decrease of ammonia production from dung. Taken together, these data showed that aluminum salt addition in poultry litter was an effective method to reduce ammonia volatilization, which may benefit to the control of ammonia production and environmental condition in hen house.

Analysis of Receptor Binding Domain on the S Protein of Porcine Epidemic Diarrhea Virus

SUN Dong-bo;;CHEN Jian-fei;SHI Hong-yan;SHEN Shi-chuan;LV Mao-jie;FAN Xiu-ping;CHEN Hong-yan;FENG Li

2009, 40(4): 528-532. doi:

Abstract ( 811 )

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Porcine epidemic diarrhea virus (PEDV) S1 gene-targeted library was micropanned three times by using porcine aminopeptidase N (pAPN) as a cellular receptor of the PEDV, and thirty phage clones were sequenced using the specific primers. The result of sequence analysis indicated that polypeptides displayed by twenty-eight phage clones were focused on amino acids 249-529 (MER) of the S protein with the exception of two meaningless sequences. The MRR recombinant protein was expressed using Bac-to-Bac baculovirus expression kit. The result of Western blot demonstrated that MRR recombinant protein could react with rabbit antisera against PEDV.

Cellular Localization and Tissue Profile of Porcine Integrin αvβ6 as FMDV Receptor

ZHAO Jian-yong;CHANG Hui-yun;DU Jun-zheng;GAO Shan-dian;SONG Shuai;FU Xiao-ping;XIE Qing-ge

2009, 40(4): 533-537. doi:

Abstract ( 758 )

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CHOK1 cell was stablly transfected by pcDNA3.1(+)β6p eucaryon expression vector, then the expressing disposition of integrin αvβ6 was detected by indirect immunofluoresence assay (IFA). Tissue profile of porcine integrin αvβ6 in porcine tongue, lip, trachea and so on, was detected by indirect immunohistochemistry assay(IHA). The monoclonal antibody against porcine integrin β6 subunit was used as first antibody, two antibodies of goat against mice IgG labelled with fluorescein isothiocyanate(FITC) and horseradish peroxidase(HRP) respectively were used as secondary antibody. RESULTS: Viridis fluorescence was appeared in cell membrane and cytoplasm of CHOK1 cell stablly transfected by pcDNA3.1(+)β6p eucaryon expression vector, buffy idio-positive reaction was appeared in cell membrane and cytoplasm of porcine tongue, lip, trachea and so on, which indicates integrin αvβ6 receptor is generally expressed in porcine tissue cell, which lay a foundation for further understanding of the roles of integrin αvβ6 in FMDV infection.

Cloning and Prokaryotic Expression of Bovine Leukemia Virus env（gp51） Gene and Development of an Indirect Enzyme-Linked Immunosorbent Assay for Detecting Antibody against Bovine Leukemia Virus

ZHOU Yi;WU Yu-shi;DUAN Hong-an;ZHANG Rui;ZHOU Zu-tao;BI Ding-ren;SHI De-shi

2009, 40(4): 538-543. doi:

Abstract ( 749 )

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The DNA of bovine leukemia virus (BLV) provirus was extracted from fetal lamb kidney (FLK) cell line persistently infected by BLV. The env（gp51）gene which encode gp51 protein was amplified by PCR. Sequencing results showed that the amplified env gene was a 828 bp fragment, the gene was inserted into the expression vector pET-32a, the recombinant plasmid was named pET-32a-gp51. A fusion protein was expressed in BL21 (DE3) that transfected by pET-32a-gp51 and induced by IPTG. The molecular weight of the recombinant protein was about 43 000 by SDS-PAGE, and the immunoreaction activity of the recombinant protein was confirmed by Western-blot, in which positive serum against BLV was used. An indirect enzyme-linked immunosorbent assay for detecting antibody against BLV was developed by using expressed protein gp51 as coating antigen. Results showed that the optimal coating concentration for gp51 is 2.19 μg·mL^-1 and the serum dilution fold is 1∶80. The positive criterion for this ELISA assay is OD450 nm> 0.451. Accuracy rate for detecting 12 standard BLV sera is 91.67%.

NF-κB Activated in PrP106-126-induced Neurotoxicity in Mouse Neuroblastoma Cells N2a

BAI Yu;ZHOU Xiang-mei;YIN Xiao-min;YANG Jian-min;ZHAO De-ming

2009, 40(4): 544-547. doi:

Abstract ( 728 )

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The neurotoxic mechanism of PrP106-126 still remains uncertain. Herein PrP106-126-induced neurotoxicity in mouse neuroblastoma cell line N2a, for the first time we found that PrP106-126 induced p65 subunit of nuclear transcription factor NF-κB translocated into the nucleus by Western Blot assay and activated NF-κB signaling pathway. Pretreatment with nuclear translocation inhibitor NF-κB SN50 attenuated PrP106-126-induced NF-κB activity, using apoptotic DNA Ladder assay and flow cytometry (FCM) assay with Annexin V-FITC/PI double-staining, we also found that attenuated NF-κB activity was accompanied by reduced apoptotic effect by PrP106-126. The results showed that NF-κB signaling pathway was involved in PrP106-126-induced neurotoxicity, and NF-κB activation showed pro-apoptotic effect in N2a cells. The in vitro study on PrP106-126-activated NF-κB signaling pathway and its pro-apoptotic effect on N2a cells were significantly important to elucidate the neurotoxic mechanism of prion.

Cloning and Genetic Analysis of the Entire Genome of A/mallard/Yancheng/2005(H4N6) Avian Influenza Virus

PENG Yi;XUE Feng;LI Yan-fang;CHEN Hao;PENG Da-xin;LIU Xiu-fan

2009, 40(4): 548-553. doi:

Abstract ( 1321 )

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An H4N6 subtype avain influenza virus(AIV), A/mallard/Yancheng/2005（H4N6）,was isolated from cloacal swab of mallard in the national protection grade in Yancheng country. The eight full-length genes， including the 5′and 3′ends of virus were amplified by using the universal primers. Amino acid sequence comparisons and phylogenetic analysis of A/mallard/Yancheng/2005（H4N6）AIV was made with other sequences available in the Influenza Sequence Database. The results showed that the motif of HA cleavage site is PEKASR， which was the characteristics of low pathogenic AIV. Both the NA and NS gene had no residue lost. The amino acid residues related to host specialty of matrix protein 2 were all avain specialty，and the 627 residue of PB2 was Glutamic acid (E). The nucleotide homology of M gene with A/duck/Yangzhou/02/2005(H8N4) AIV was 99.4%, the PA gene had genetically close relationships with that of A/duck/Jiangxi/1286/2005(H5N2) AIV, these results suggest that both M and PA gene had circulated in the domestic duck and involved in gene reassortment.

Determination of the Outer Membrane Protein (OMP) Patterns of Pathogenic E. coli Isolated from Ducks

YU Xue-hui;CHENG An-chun;WANG Ming-shu;TANG Cheng;WANG Yuan-wei;WANG Ying;YUE Hua;ZHANG Huan-rong

2009, 40(4): 554-560. doi:

Abstract ( 822 )

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The outer membrane patterns of 130 pathogenic E. coli strains isolated from ducks of 3 provinces or municipal cities in southwest China were determined in this study. Among these 130 strains, 53 isolates belonged to prominent serotype O93, O92, O78 and O76, while 77 strains were serotype O46 and other serotypes. There were 4 different OMP patterns among all strains tested. 62.3% (81/130) of strains that belong to 29 serotypes were identified to be OMP-1， including O93，O92，O78 and O76, suggesting that OMP-1 was the main OMP pattern. Subsequently, a pair primers for amplification of ompA gene was designed according to sequence of the E. coli K-12 available in GenBank. The ompA gene were then amplified and sequenced from 9 prominent serotype strains and 1 serotype O46 strain. Sequence analysis revealed that the ompA gene of all 9 prominent serotype strains was 1 171 bp long and composed of only one large open reading frame (ORF) with a size of 1 053 bp, which encoded a protein consist of 350 amino acids, proompA. The first 21 amino acids encoded a signal peptide and the mature OmpA protein was composed of 329 amino acids. The ORF of ompA gene from O46 serotype strain was 1 041 bp in length with a 12 bp depletion at the site of 400-411 bp compared to other serotype strains, and the mature OmpA protein contained 325 amino acids. It was also found that the nucleotide sequences of ompA genes hared high homology with an identity ranging from 95.8% to 100% among 10 pathogenic E.coli strains from ducks.

Influence of Amino Acids on Traps Forming of Nematode-trapping Fungus—Arthrobotrys oligospora

DENG Chang-sheng;WANG Rui;YANG Xiao-ye;HAN Hai-bing;YANG Lian-ru;LIU Zhen-lian;ZHANG Xiao-dong

2009, 40(4): 561-565. doi:

Abstract ( 703 )

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In order to study the effects of amino acids on the nematode-trapping organs forming of Arthrobotrys oligospora, 0.5 mL solutions of amino acids（0.5, 0.05 and 0.005 g·L^-1）were spreaded over the CMA with A. oligospora in different growing stages respectively, then they were incubated in 25 ℃. The results showed that different amino acids had different effects on the traps forming. Among all of the amino acids tested, L-Phenylalanine, L-Methionine and DL-Valine of 0.05 g·L^-1 solutions had the best effects on inducing traps, and L-Threonine, DL-Phenylalanine, L-Isoleucine, L-Tryptophanea, DL-Leucine, L-Leucine and L-valine had better effects than the others, but worse than the former ones. It was easier for young hyphae to produce traps, which were similar in trapping and killing nematodes with those induced by nematode larvae, than the older ones when they were induced by amino acids. All those showed that the nematode-trapping organs of A.oligospora could induce not only by nematodes but also by amino acids, and they could also trap and kill larvae of nematodes effectively, which was very useful and important to study the biochemical mechanism of nematode-trapping fungi.

Effects of Two Histamine Receptor Antagonists on Lung Injury Induced by Endotoxin in Chicken

OU De-yuan;FU Xue-zhi;GAO Ming-yu;XU Le-ren;YAO Hong-yan

2009, 40(4): 566-571. doi:

Abstract ( 720 )

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The experiment was conducted to investigate the effects of two histamine receptor antagonists, chlorphenamine maleate and cimetidine, on acute lung injury induced by endotoxin in chickens. Methods: One hundred and twenty birds aged 20 d were randomly divided into five groups. (1) Endotoxin group (EG): The normal saline was given twice on morning and afternoon, and endotoxin was given on noon;（2）Chlorphenamine maleate group (CM): The chlorphenamine maleate was given twice on morning and afternoon, and endotoxin was given on noon; (3)Cimetidine group (CMT): The cimetidine was given twice on morning and afternoon, and endotoxin was given on noon;(4)Chlorphenamine galeate and cimetidine group (CMC): The chlorphenamine galeate and cimetidine was given twice on morning and afternoon, and endotoxin was given on noon;(5) Control group(CG): The normal saline was given twice on morning and afternoon. Seven chickens were randomly taken from each group on the 5th, 10th, 15th day. The birds were weighed and killed. The lung was weighed, then it was fixed in neutral buffer formalin or stored in ultra cold freezer. The histamine was measured by OPT. Paraffin sections were made. Hematoxylin and eosin staining was used. The score criterion of pathology changes were measured. Results: (1) The contents of histamine in lung of CMC was lower than that of EG, CM and CMT. At the 5th day，CMC was significantly lower than that of CM and CMT (P<0.05).At the 10^th day,EG was significantly (P<0.05) higher than that of CMC and CG. At the 15^th day, EG was significantly higher than that of CMC (P<0.01), CM and CG (P<0.05). (2) The index of lung in CM and CG was significantly less (P＜0.05 ) than that in EG at the 10th day， the index of lung in CM, CMC and CG was significantly less (P＜0.05 )than that in EG at the 15^th day. (3) The degree of lung injury of EG was significantly severer than other groups all the time (P<0.01), CMC was significantly lower than that of CM and CMT at the 5^th day (P<0.01)， but CMC was significantly lower than that of CM (P<0.05) and CMT (P<0.01) at the 10^th, 15^th day. Those data indicated that histamine play roles in the endotoxin-induced lung injury through H₁ and H₂receptors, but mainly through H₁ receptor in chicken.

Effect of High Copper on the Antioxydic Function of Kidney in Ducklings

CUI Wei;PENG Xi;ZHAO Li;YANG Fan;CUI Heng-min

2009, 40(4): 572-576. doi:

Abstract ( 728 )

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The experiment was conducted with the objective of examining the effect of dietary high copper on the antioxydic function of kidney in ducklings. 360 one-day-old Tianfu meat ducklings were randomly divided into six groups and feed on diets as follows: Controls (Cu 8 mg·kg^-1) and high copper (Cu 100 mg·kg^-1，high copper groupⅠ；Cu 200 mg·kg^-1，high copper groupⅡ；Cu 400 mg·kg^-1，high copper group Ⅲ；Cu 600 mg·kg^-1，high copper group Ⅳ；Cu 800 mg·kg^-1，high copper groupⅤ) for six weeks. Compared with those of control group, the renal free fatty acids, malondialdehyde and hydroxy radical contents in high copper groups Ⅱ,Ⅲ, Ⅳ and Ⅴ, and the renal cuprozincsuperoxide dismutase(CuZn-SOD) in group Ⅴ were increased (P<0.01 or P<0.05). Also, the serum copper-protein (CP) activities in high copper groups Ⅲ, Ⅳ and Ⅴ and the serum CuZn-SOD activities in group V were lower (P<0.01 or P<0.05 ) than that in control group.The results showed that dietary copper exceed 200 mg·kg-1 impaired renal antioxydic function, and the renal function was finally depressed.

Dynamic Changes of Renal Function and Urinary Sediment during the Onset of Urolithiasis in Goats

SUN Wei-dong;WANG Jin-yong;YAN Li-hua;WANG Xiao-long

2009, 40(4): 577-583. doi:

Abstract ( 652 )

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The urolithiasis of 6 wether lambs was successfully induced by a new mode of feeding with cottonseed meal diet and adding MgO in drinking water, and the changes of urinary sediment and renal function were evaluated in different experimental phase of urolithiasis. The experiment was divided into five phases, the end of pre-testing designed as phase Ⅰ, days 15-60 of experiment as phase Ⅱ, pre-dysuria after adding MgO in drinking water as phase Ⅲ, the initial stages of dysuria as phase Ⅳ, 2-5 days after dysuria as phase Ⅴ. The results showed that: (1) Crystals emerged largely with coffinlid-shaped and different size radicel crystals in phases Ⅱ and Ⅲ in urine of goat after cottonseed meal and rice straw diet feeding, and basically disappeared in phase Ⅳ, but small crystal could be found again in urine of phase Ⅴ with a significant lower quantity than that in phases Ⅱand Ⅲ (P<0.05). (2) Small amounts of tubules epithelium was observed in urine of goat in phases Ⅰ and Ⅱ. At the initial phase of occurrence of urolithiasis induced by MgO treatment (phases Ⅲ and Ⅳ)，cast-off cells of tubules epithelium and pelvis epithelium with small amounts of cast were observed in urine from bladder. Several days after the occurrence of urolithiasis (phase Ⅴ) amounts of cast-off cells and casts got a further increase. (3) Urea in plasma got a significant increase (P<0.05) in phases Ⅱ, Ⅲ, Ⅳ and Ⅴthan in phase Ⅰ. Plasma creatinine was found no significant change in phases Ⅰand Ⅱ, and significant increase (P<0.05) in phases Ⅲ, Ⅳ and Ⅴ than in phases Ⅰand Ⅱ. The ration of plasma urea to plasma creatinine increased significantly (phase Ⅱ) after cottonseed meal and rice straw diet feeding but decreased significantly (phases Ⅳ and Ⅴ) after the occurrence of lithogenesis. Urine protein got a significant increase (phases Ⅱ and Ⅲ) but decreased significantly at the initial stage of lithogenesis (phase Ⅳ)，then a significant increase was found several days later (phase Ⅴ) of anuresis. Conclusion: High intake of magnesium could provoke the shedding of nephric epithelium, this condition contributed to the change of renal function. At the same time the cast-off cells and urinary protein also offered a convenience of calculous nucleation and promoted the occurrence of urolithiasis in goat. These changes could be helpful to understand the question how does crystal convert to calculi.

Dynamic Effect of Tiletamine on AC Activity and cAMP Content in Different Brain Regions of Rats

FAN Hong-gang;LU De-zhang;ZHANG Jian-tao;HU kui;WANG Hong-bin

2009, 40(4): 584-588. doi:

Abstract ( 1124 )

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To observe the dynamic effects of tiletamine on AC activity and cAMP content in different brain regions of rats, 168 SD rats were randomly divided into two groups, and the activity of AC and the cAMP content were measured respectively. Every group was divided into control group, high dose group(ip tilelamine 60 mg·kg^-1) and low dose group(ip tilelamine 30 mg·kg^-1). The group was divided into three subgroups of maintenance of anesthesia, recovery from anesthesiaⅠand recovery from anesthesiaⅡ according to their anesthesia state. The activity of AC and the cAMP content were measured by radioimmunoassay (RIA). The results of experiment showed that in the high and low dose maintenance of anesthesia groups, the AC ctivity in cerebral cortex and thalamus were obviously increased, and the cAMP content in abovementioned regions were also obviously increased(compared with the control group, P<0.05). In the recovery from anesthesiaⅠgroups of high dose group and low dose group, the activity of AC and the cAMP content were decreased at different degree, and the high dose group was decreased obviously(compared with maintenance of anesthesia group, P<0.01). In the recovery from anesthesiaⅡ, they were obviously decreased(compared with maintenance of anesthesia group, P<0.01). Two dose groups have insignificant effect on the AC activity and the cAMP content in hippocampus, brain stem, and cerebellum. The results suggested that the AC activity and the cAMP content might play an important part in the molecular mechanisms of general anesthesia mediated by tiletamine. The anesthetic action of tiletamine might relate to enchance the activity of AC and increase the cAMP content in the cerebral cortex and thalamus.

The Cloning, Sequencing, and Protein Structure Analysis of Type ⅠSmall Peptide Transporter in the Gastrointestinal Tissue of Bovine

LIN Xue-yan;LI Yan;JIANG Yun-liang;SU Peng-cheng;WANG Yun;WANG Zhong-hua

2009, 40(4): 589-594. doi:

Abstract ( 1333 )

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cDNA was synthesized using the whole RNAs extracted from bovine omasal flakes in the present study. The cDNA of bovine type Ⅰ small peptide transporter (BPepT1) was amplified by RT-PCR using primers designed according to the published sequences of human, mouse, rabbit, and sheep PepT1 in GenBank. The amplified segment was sequenced after purifying and cloning. After comparing with the published PepT1 sequences of other animals, the 1 566 bp sequenced segment (DQ309694) was a sequence encoding the peptide chain from 3rd to 10th transmembrane structural domains of BPepT1. Homologous analysis indicated that the sequenced segments of BPepT1 had 82.89%, 80.14%, 78.93%, 96.04%, 83.08%, 63.90%, 85.66% nucleotide identity and 81.45%, 79.62%, 76.25%, 94.06%, 81.49%, 61.41%, 83.56% amino acid identity with the corresponding segments of human, rat, rabbit, sheep, dog, chicken, and pig PepT1, respectively. Comparison of the PepT1 amino acid sequences encoded by the sequenced segments of eight animal species (bovine, human, mouse, rabbit, sheep, dog, chick, and pig) indicated that PepT1 was a highly conservative protein during evolution, and the amino acid sequences in the transmembrane structural domains were more conservative than those in the outside domain sequences. The amino acid identities of the 12 structural domains were all above 90%, and the identities from 3rd to 10th domains were 90.97%, 91.07%, 97.62%, 93.45%, 94.05%, 91.67%, 91.45%, and 95.83%, respectively. The amino acid identities of the outside domain regions ranged from 70% to 99%, and the amino acid identities of the 3-4, 4-5, 5-6, 6-7, 7-8, 8-9, and 9-10 outside-domain regions were 77.63%, 98.86%, 86.72%, 90.13%, 91.25%, 88.33%, and 71.74%, respectively. The outside domain region between the 9-10 trans-membrane domains is a big extracellular loop, which may be an important structure in substrate recognition by PepT1. The amino acid identity in the region between bovine and chick was quite and only 29.3%, but the amino acid identities between bovine and mouse, dog, human, rabbit, pig, or sheep were 65.17%, 71.64%, 67.16%, 60.20%, 73.13%, and 88.56%, respectively. Predicated from the published consensus amino acid sequences, there were six N-linked glycosylation sites, three cAMP-dependent protein kinase (PKA) sites, and one protein kinase C (PKC) site in the amino acid sequence encoded by the sequenced segment of BPepT1.

Molecular Cloning and Sequence Analysis of the Amino Acid Transporter ^b0,+AT-2 of Pigs

ZHI Ai-min;FENG Ding-yuan;ZHOU Xiang-yan;HUANG Zhi-yi;ZOU Shi-geng;WANG Xiu-qi;ZUO Jian-jun;YE Hui;WANG Xiao-lan;LIU Zhun

2009, 40(4): 595-601. doi:

Abstract ( 1166 )

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The cloning, sequencing of porcine b^0,+AT transcript variant 2 was described in this study. Based on the sequences of human and mouse b^0,+AT published on the NCBI, we identified a partial cDNA using the primers designed after bioinformatics analysis. Using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine b^0,+AT-2 was sequenced. The porcine b^0,+AT-2 cDNA was 1 488 bp long, including 90 bp 3′UTR and 126 bp 5′UTR, and CDS encoded 423 amino acids. The b^0,+AT-2 protein sequence was found to be 75.1%，71.9% and 86.6% identities with human b^0,+AT, mouse b^0,+AT and swine b^0,+AT-1.

HA and NA Gene Sequence Analysis of Ten Avian Influenza Virus Subtype H9N2

XU Dong;ZHANG Xiu-mei;HUANG Yan-yan;HU Bei-xia;LI Jian-liang;CUI Yan-shun

2009, 40(4): 602-608. doi:

Abstract ( 1214 )

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In order to explore the genetic mutations of the hemagglutinin gene and the neuraminidase gene and the law of molecular epidemiology of H9 subtype avian influenza in China, ten avian influenza virus subtype H9N2 were isolated from some poultry yards in Northern China, the whole HA and NA cDNA fragments of them were amplified by reverse transcription polymerase chain reaction, with primers specific to HA gene and NA gene. After cloning and sequencing, gene sequences were analyzed on homology and heredity evolution. The sequence analysis showed little degree of variation, the various types and the high rate of mutation. They belonged to a branch of the CBJ194-like in the phylogenetic tree. The nucleotide homology of the HA gene to the CBJ194 was from 88.8% to 95.8%. The nucleotide homology of the NA gene to the CBJ194 was from 93.4% to 97.6%, so they might come from the same ancestor AIV strain. Because of the gene mutation, the new potential glycosylation site appear in the HA gene of the ACHB101 and a stenoplastic receptor-binding site have changed in ACHN102. In the deduced amino acids sequence, the 63, 64 and 65 sites lost T, E and I, which cause loss of the 61 glycosylation sites. Otherwise, there was conspicuous mutation in the acetylneuraminic acid adsorption sites. These findings may be fruitful for making further study of genetic mutations of AIV in the field and improving the control strategies.

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