Acta Veterinaria et Zootechnica Sinica

Construction of Eukaryotic Expression Vector Containing Human Cardiac α-actin Promoter and It’s Application

2006, 37(11): 1093-1098. doi:

Abstract ( 1288 )

PDF (595KB) ( 620 )

Related Articles | Metrics

Human cardiac α-actin promoter obtained from the human heart genome by polymerase chain reaction(PCR) was ligated into the same fragment of pEGFPN1（moved CMV promoter by AseI/Bgl II）.The constructed expression vector, named α-actin-pEGFP-N1 which contains neor gene and GFP gene as positive selection markers,was transferred into mice ES cells by Lipofectamine 2000 It’s found that DNA concentration should be 3.0-5.0μg/mL, transfer time was 2-3h, 200μg/mL G418 was used as perfect working concentration. Fluorescent observation and immunocytochemistry assay (PCNA and antibody of α-actin) demonstrated the transfected cells show high levels of α-actin and GFP expression, basically maitain its natural characteristic compared with wild mice ES cells. The results suggest that human cardiac αactin promoter may be useful in the study of mice ES cells specific differentiation into cardiomyocytes.

Construction of Resource Populations for Mapping Chicken Growth and Meat Quality Traits

2006, 37(11): 1099-1106. doi:

Abstract ( 705 )

PDF (618KB) ( 615 )

Related Articles | Metrics

F₂ resource populations, consisting of 4 branch families, were constructed from reciprocal out-crossings of Xinghua chickens (Line A), White Recessive Rocks (Line B) and Taihe Silkies (Line C), in which the mating of male line A and female line B, male line B and female line A, male line A and female line C, male line C and female line A produced full-sib and half-sib individuals, with outbred F-2 design. From then growth performance and meat quality traits in F₂ individuals were measured and depicted by stages. The analysis of phenotypic values indicated that the measured traits on growth performance and meat quality were segregating well in F2 individuals. The population size is large enough, and the resource population can correct maternal and environmental effects. It can promise the scale and density of single nucleotide polymorphisms and the accuracy of QTL mapping. Therefore, the construction of resource populations achieved the expected aim.

Study on Molecular Genetic Diversity of Native Duck Breeds in China

2006, 37(11): 1107-1113. doi:

Abstract ( 674 )

PDF (721KB) ( 634 )

Related Articles | Metrics

Twenty four Chinese native duck breeds were studied to estimate genetic diversity and genetic structure using microsatellite markers. The genetic relationships between breeds were analyzed in combination with their geographic distribution. A total of 28 microsatellite markers were amplified .All breeds exhibited genetic diversity, through the estimation of polymorphic information content (PIC) and mean heterozygosity (H) index. The mean genetic diversity of the populations was 0.569, while the PIC values for Chinese native duck breeds showed a relatively high level, except APL23 and APL79 were similar (0.414) and of medium polymorphic. The Neighbor-joining was used to calculate the genetic distances. Cluster analysis divided the twenty-four populations into five groups, and the genetic relationships among the populations had obvious association with their phylogenetic relationship with the historical relations and geographical distribution. The development history of native duck breeds in China seemed to have an effect on their genetic structure.

Analysis on Genetic Variation of Adipocyte Fatty Acid Binding Protein Gene in Different Chicken Breeds

2006, 37(11): 1114-1117. doi:

Abstract ( 670 )

PDF (698KB) ( 730 )

Related Articles | Metrics

The genetic variation of Adipocyte fatty acid binding protein gene on the sequence sites of 85 and 1 805 in 96 chicken including Youxi Partridge, Luyuan and Recessive Whitefeather chicken were detected by PCR-RFLP and DNA sequence analysis. The results showed as follows: 1）All these chicken breeds had genetic variation in A-FABP gene on sites 85 and 1 805. Recessive Whitefeather chicken presented mostly Cc genotype while other two breeds presented mostly cc genotype. The allele c had high frequency in three chicken breeds and in Youxi partridge chicken was the highest and Recessive Whitefeather chicken was the lowest. 2）The genetic variation on 85 site had no effect on amino acid sequence of A-FABP and 1 805 site had effect on amino acid sequence of A-FABP: Pro→Ser which suggested that the chicken quality may have relation to A-FABP’s expression possibly controlled by genetic variation.The research built the base for further analysis on relation between genetic variation of A-FABP and the IMF quantity, and molecular marker’s application in breeding.

Sequencing Cytochrome b Gene of Mitochondrial DNA in Yak and Researching Its Origin and Taxonomic Status

2006, 37(11): 1118-1123. doi:

Abstract ( 1110 )

PDF (768KB) ( 757 )

Related Articles | Metrics

There still exists remarkable difference on yak’s taxonomic status in Bovinae. Primers designed according to the Mitochondrial gene sequences of Bos taurus reported was used to amplify and sequence yak’s Cytochrome b gene, and the whole sequence of Cytochrome b gene was obtained .Using Ovis aries as outgroup taxa, the Phylogeny about the representative species of Bovinae was analyzed .The results showed that among different species the transition/transversion(Ts/Tv) of Cytochrome b gene was 4.9, suggesting that the mutation was not saturation; the percentage nucleotide sequence divergence between yak and Bovinae was 8.0%~8.6%，which was more than that of yak and Bison bison; Phylogeny analysis found that Poephagus grunniens and Poephagus mutus clustered first of all, then gathered with Bison bison, indicating there was higher genetic comparability than that of Bos, the results sustained the idea Poephagus grunniens and Poephagus mutus owned one ancestor —primitive yak;The approximate divergence time between these two species was 0.55 million years. The data also supports the point that yak is classified as Poephagus of Bovinae, Poephagus including two species: Poephagus grunniens and Poephagus mutus.

Polymorphism Research of Microsatellite DNA in Nine Sichuan Black Goat Breeds (Populations)

2006, 37(11): 1124-1129. doi:

Abstract ( 1374 )

PDF (836KB) ( 713 )

Related Articles | Metrics

Genetic diversity for Sichuan black goat populations, including Jintang black goat, Lezhi black goat, Jianchang black goat, Hejiang black goat, Jiangan black goat, Zigong black goat, Jialing black goat, Yingshan black goat and Baiyu black goat, were estimated using ten microsatellite makers by PCR, polyacrylamide gel electrophoresis and silver staining of Sanguintti method. The results showed that ten microsatellite loci analyzed were highly polymorphic loci in nine populations. The average PIC, H and Ne of Jintang, Lezhi, Jianchang, Hejiang, Jiangan, Zigong, Jialing, Yingshan and Baiyu black goat were 0.724 4/0.763 2/4.483 8, 0.750 3/0.782 7/4.878 2, 0.735 2/0.773 9/5.017 6, 0.770 2/0.800 7/5.286 1, 0.740 2/0.777 1/4.576 8, 0.746 5/0.784 6/ 4.855 6, 0.751 1/0.788 9/ 4.959 1, 0.760 4/0.790 5/4.886 9 and 0.697 0/0.741 7/4.272 8. Jialing and Yingshan black goat was one cluster (D=0.386); Zigong and Jiangan black goat was separated one cluster (D=0.428) firstly, then Hejiang black goat joined this cluster (D=0.456); Jingtang and Lezhi was one cluster (D=0.437); Baiyu and Jianchang black goat was one cluster. At last, the four clusters build up a big cluster.

Sequencing and Analysis of Luteinizing Hormone Beta Gene from Nanjiang Huang Goat

2006, 37(11): 1130-1134. doi:

Abstract ( 640 )

PDF (866KB) ( 544 )

Related Articles | Metrics

According to the LHβ gene sequence of cow, sheep and pig in GenBank, the primers were designed and partial sequence of LHβ gene from 35 Nanjiang Huang goat blood samples were amplified and sequenced by PCR method (GenBank No:AY853264). Furthermore, phylogenetic tree of different species were conducted using the LHβ partial coding sequence. The result showed that the 929bp partial sequence of LHβ gene included 5′flanking region (136bp), two introns (297bp and 235bp), two exons (26bp and 168bp) and partial sequence of exon3 (67bp). Exon sequence coded 83 amino acids except the 11bp nucleotide at the beginning of 1st exon ,which was the 5′untranslating region(5′UTR), while the first 20 amino acids were the signal peptide. LHβ gene was rich with GC. Eight nucleotide mutations were detected and two of them existed in exon were synonymous mutation. Phylogenetic relationship was not in accord with the actual species evolution process, which maybe caused by the different GC proportion of LHβ gene among species. It was the first time to report goat LHβ gene sequence which established the foundation of the further research about genetic principle of goat reproduction.

Nae I Polymorphism of PRLR Gene and Its Effect on Reproductive Traits in Minpig, Landrace and Their F1 Crossbreed Sows

2006, 37(11): 1135-1140. doi:

Abstract ( 1024 )

PDF (949KB) ( 697 )

Related Articles | Metrics

The prolactin receptor (PRLR) plays a key role in many reproductive pathways of animals. It is probable that the change of the structure of PRLR gene will lead to variation of PRLR function, and hence affect reproductive phenotype. In this experiment, the Nae I polymorphism of PRLR gene and its effect on reproductive traits in Minpig, Landrace and their F1 crossbreed sows was studied by sequencing and PCRRFLPs. A mutation of A to G (1 789) that resulted in a change of amino acid (S→G), i.e. Serine → Glycine was detected in cDNA sequence (+1 628+1 878) of PRLR gene. The frequencies of genotypes and alleles for Nae I polymorphism site were significantly different in three populations (P<0.01). The results of correlation analysis indicated that PRLR gene was closely associated with several reproductive traits examined. As regards Minpig, only two genotypes were detected, sows with AB genotype had significantly heavier litter weight at birth and higher piglets uniformity on day 21 than sows with AA genotype (P<0.05). For Landrace, sows with AA genotype have a greater litter weight on day 21 than sows with BB genotype (P<0.05). In the F1（L×M）crossbreed population, piglets uniformity on day 21 was significantly higher for sows with BB genotype than sows with AB and AA genotypes (P<0.05).

Research on the Six Fluorescent Protein Expression in the Fibroblast Cells of Mongolia Sheep

2006, 37(11): 1141-1148. doi:

Abstract ( 798 )

PDF (1069KB) ( 629 )

Related Articles | Metrics

To study the exogenous gene space-time expression, the state of positive cell growth and reduplication, apoptosis, survival percentage，and to optimize the method improving the transfection efficency, we used lipofectin method to transfect six fluorescent protein genes: pEGFP-C1、pEGFP-N3、pECFP-N1、pECFP-mito、pEYFP-N1 and pDsRed1-N1 into the in vitro cultured fibroblast cell of Mongolia sheep. The results showed: 24、48 and 72h after transferring, the expression efficiency of six kind of fluorescent protein genes were between 12.3％～63.8％.Multiple comparison analysis indicated significant difference in six groups. 24h after transferring, the fluorescence could be observed in cytoplasm and nucleus welldistributed except cryptomere vesicle ; 48h after transferring, EGFP、ECFP, and EYFP gene still expressed steadily. Granula could be observed in the region of cell nucleus. DsRed gene mainly expressed some granulaexpression product surrounding nuclear membrane. In optimized condition, there were no significant effect（P>0.05）on apoptosis ratio, positive cell shape, growth and reduplication state comparing with control group. The research is important to genetic mark, nuclear transplantation and transgenic animal clone etc.

Effects of Leptin on the Expression of Ob-Rb mRNA in the Cultured Adipocytes of Newborn Calf

2006, 37(11): 1149-1153. doi:

Abstract ( 658 )

PDF (989KB) ( 523 )

Related Articles | Metrics

A single factor duplicate test was designed to investigate whether additional leptin(5 ng/mL·12 h) in adipose cell cultured medium at 12,24,36,48 and 72 hours(12 hour each)has a role on ObRb mRNA expression by competitivePCR. The result showed that the quantity of ObRb mRNA in adipose cell cultured for 12～24 h was significantly higher in the treated groups than that of control group（P<0.01）.Following gradual decreasion ,the expression of ObRb kept a stable level from 48 to 72 h（P>0.05）.It considered that leptin could accelerate the expression of ObRb in the cultured adipose cells with giveen time.

hCG Regulating the Expression of StAR through ERK1/2 Pathway

2006, 37(11): 1154-1159. doi:

Abstract ( 675 )

PDF (1091KB) ( 579 )

Related Articles | Metrics

Leydig cells from 2-3 week old piglets were used to study the mechanism of hCG regulating Steroidogenic acute regulatory protein(StAR). The results of this experiment were as shows : (1)Both of hCG and 8-Br-cAMP could enhance both the expression level of StAR protein and that of its mRNA ,increase active ERK1/2 clearly depending on stimulating time. However, the effect of 8-Br-cAMP is faster than that of hCG. (2) While appending the inhibitior of PKA ( PKI) to leydig cells in culture , both of the expression level of StAR protein and that of its mRNA began to drop significantly and the activity of ERK1/2 was inhibited but the level of StAR mRNA could still be detectable.(3) While appending the inhibitior of MAPK (PD98059), the expression level of StAR protein mRNA dropped significantly .These results inferred that at the beginning of hCG stimulating, hCG increased the level of StAR protein by cAMPPKA. With the increase of time, hCG increased the level of StAR protein through cAMPPKA ERK1/2

Effect of Inhibin Gene Immunization Fused with HBsAg on Rat Reproductive Performance

2006, 37(11): 1160-1166. doi:

Abstract ( 632 )

PDF (1036KB) ( 678 )

Related Articles | Metrics

A total of 90 rats were divided into five groups (n=18) and immunized i.m. with 10, 50, 100μg inhibin eukaryotic expressing plasmid fused with HBsAg gene (pCIS), 50μg vector (pcDNA3.1) or 100μL 0.85% saline. A booster was given 20 days later, and 12 of rats in each group were given twice boosters. The results showed that the boosters raised the level of antibody against inhibin. There were 6.9 and 7.5 more mature follicles in 100μg group after twice and three times immunizations of pCIS than that in the control(P<0.05). The mature follicles after three times immunizations were remarkably more than that after twice immunizations(35.2±2.73 Vs. 31.0±0.92，P<0.05). After three times immunizations of pCIS, the placentas and litter size in total trials were more than that in the control (P>0.05), and positive group more than negative group (P<0.05). After three times immunizations , the correlation coefficient between the positive antibody and the mature follicles was 0.45（P>0.05）, while it got to 0.77（P<0.05）between the positive antibody and the placentas. The level of FSH on estrus and after parity in total trials was higher than that in the controls, which in positive rats was higher than that in negative rats, especially after twice immunizations (P<0.01). In conclusions, inhibin gene immunization can stimulate follicular development of rats and raise the level of plasma FSH, which set up the basis of inhibin gene immunizations on large animals.

The Study of Cryopreservation Technique of Biopsied Mouse and Bovine Embryos

2006, 37(11): 1167-1172. doi:

Abstract ( 667 )

PDF (1004KB) ( 741 )

Related Articles | Metrics

Cryopreservation technique of biopsied mouse and bovine blastocysys were also studied. As a result,(1) The biopsied mouse embryos were freezed with 10% glycerol and 10% glycol respectively and, as comparison, the biopsied mouse embryos were freezed with both of the freezing solution added 10% polysaccharid and 10%sucrose, the development rate of frozenthawed mouse embryos were 56.9%(259/455), 81.8%(317/390), 84.8%(540/738), 59.5%(308/518) respectively. (2)The development rate of frozenthawed mouse embryos could not be improved when the biopsied embryos were cultured in vitro shortly (about 0-4 hours) after sampling. (3)Being suffocated 2 minutes at -100 ℃--110 ℃ after sampling and fast cryopreservation with frozen solution which include 25% glycerol and 0.25 mol/L sucrose and 20% blood serum, the development rate of muse embryos was 733%, the pregnancy rate of fast frozenthawed bovine embryos after sampling was 57.1% (4/7).

Isolation, Purification and Culture of Spermatogonia in Chicken

2006, 37(11): 1173-1178. doi:

Abstract ( 700 )

PDF (1077KB) ( 692 )

Related Articles | Metrics

The study was carried out on isolation, purification and culture of spermatogonia gained from chicken testicular tissues. After enzymatic digestion, the single cell suspension was treated by percoll density gradient centrifugation to obtain high purity’s spermatogonia. The results showed that ①After percoll gradient centrifugation, the purity of spermatogonia gained from 6d-old-cock was 10.2%, 5.7% and 2.5% higher than the purity from 13d-old-cock, 15d chicken embryo and 19d chicken embryo. ②By further using adhesion purification, 82% of spermatogonia was also obtained, which was 15.6% higher than the purity of only using percoll density gradient centrifugation. ③Before and after percoll density gradient centrifugation, the different methods for cultivation spermatogonia in vitro were also compared. Results showed that: in the condition of no CEF feed layer existed, the adhesion time and survival time of spermatogonia before percoll density gradient centrifugation was earlier and longer than the time after percoll density gradient centrifugation.

Study on Requirement of Copper for Beef Cattle under Different Dietary Levels of NPN and Sulfur

2006, 37(11): 1179-1183. doi:

Abstract ( 672 )

PDF (1066KB) ( 527 )

Related Articles | Metrics

Crossbred (Limousin × Lu-xi) beef cattles, fitted with permanent cannulas in the rumen, and catheter into jugular vein, were used in the study. The experiments were divided into three stages according to three dietary levels of NPN and sulfur (0％ and 0.150%，1％ and 0.225%，2％ and 0.300%), each level corresponded with three concentrations of copper that were 10, 25 and 50 mg/kg in the diets, and were conducted in Latin square designs to access the effects of dietary NPN and sulfur on metabolism of nutrients, the bioavailability and the requirement of copper in beef cattles. The results indicated that plasma copper concentration and activities of Cp or SOD increased (P<0 .05)，but the synthesis of ruminal MCP reduced with increasing dietary copper in the same level of NPN and sulfur in the diets. In the presence of high dietary NPN and S concentration, the bioavailability of dietary copper declined，but the synthesis of ruminal MCP increased (P< 0.05). It was suggested that the optimal requirements of copper were 10, 10 and 25 mg/kg for cattles fed diets containing urea and sulfur 0% and 0150%, 1% and 0.225%, 2% and 0.300% respectively.

Effects of Curcuma zedoaria Rosc. Polyszccharide on Antioxidant Status and Immunity Function in Chickens

2006, 37(11): 1184-1188. doi:

Abstract ( 666 )

PDF (1185KB) ( 726 )

Related Articles | Metrics

Effects of Curcuma zedoaria Rosc. polyszccharide on the antioxidant status and the immunity function in chickens were studied in the experiment. One hundred and sixty 7dayold chickens were randomly divided into four groups,control group(Ⅰ),Cyclophoshamide group(Ⅱ),Cyclophoshamide plus curcuma zedoaria Rosc group(Ⅲ),Curcuma zedoaria Rosc.group(Ⅳ). The phagocytosis ability of blood corpuscle, SOD activity and LPO content of blood serum, and the chickens’ indexes of bursa F and spleen were determined in the blood of each chicken in every group. At the same time, we also observed the structures of bursa F and spleen. The results showed that Curcuma zedoaria Rosc. polyszccharide had an evident effect in eliminating ·OH and O₂- , and the biggest eliminating ratio was 88.91% and 98.5% respectively; SOD activity and LPO content were significantly changed, SOD activity was enhanced 145% and LPO content reduced 40.95%, maximally, comparing with the control group; The weights of all chickens were increased quickly from the second week after treatment, the biggest increase weight was 48.75 g per week; The phagocytosis ratio and index of blood corpuscle, weight and growth speed of bursa F and spleen in group of Curcuma zedoaria Rosc. polyszccharide were increased; The values of antioxidant and immunity functions in the group fed with the cyclophoshamide added Curcuma zedoaria Rosc. polyszccharide were similar to the control group, which it showed that the injury to immune organs by cyclophoshamide could be made up by Curcuma zedoaria Rosc. polyszccharide. All in all, Curcuma zedoaria Rosc. polyszccharide had a antioxidant ability in some degree, could strengthen body immunity ,increase growth performance of chickens, repair injury of tissue.

Development of SYBR Green I Reverse-transcription Polymerase Chain Reaction to Detect Avian Influenza Virus

2006, 37(11): 1189-1193. doi:

Abstract ( 1351 )

PDF (1108KB) ( 764 )

Related Articles | Metrics

A pair of primers were designed for detection of avian influenza viruses(AIV). 4 isolates belonging to different subtypes were used to infect MDCK cells, then total RNA were extracted from supernatant fluids after infecting 6，12，24，48，72 h. Throat, tracheal swabs samples from chicken were extracted too.SYBR Green I RT-PCR of M gene was done with Rotor Gene 3000 PCR amplifier. We attempted to establish a SYBR Green I RTPCR method to detect AIV.At the same time, the results of SYBR Green I RT-PCR were compared with those of conventional RT-PCR and virus titration.The results showed that the assay was sensitive, specific and rapid enough to detect AIV RNA genome in cell culture supernatant and swabs samples, while other viruses were not detected. As such, the described method is reliable and provides faster detection of AIV than other conventional methods.

Expression and Identification of the Protein Encoded by a Novel Gene（GP29）from Hyalomma asiaticum Tick Gland Salivary

2006, 37(11): 1194-1197. doi:

Abstract ( 652 )

PDF (1135KB) ( 582 )

Related Articles | Metrics

The open reading frame of GP29, a gene expressed only in the salivary glands of the fed adult tick H.asiaticum， was cloned into expression vector pGEX-4T-1. A novel protein of approximately 53 ku，being in agreement with that predicted from the translated cDNA sequence，was expressed by the Escherichia coli （BL21 strain）transformed with p29-pGEX-4T-1. The peptide mass mapping of the protein and the peptide sequencing of the “1381.7511 m/z” peptide suggested that the protein in the 53 ku band on SDS-PAGE was the GST-fusion protein expressed by the GP29 gene . Comparison of the sequence of the protein to that of other proteins in the Swiss PROT data base revealed no significant homology，indicating that it is a novel bioactive molecule.

Expression of Swine Hepatitis E Virus ORF2 and Development of ELISA Based on the Expressed Protein

2006, 37(11): 1198-1201. doi:

Abstract ( 1199 )

PDF (1125KB) ( 663 )

Related Articles | Metrics

To develope a method to detect the antibody of swine hepatitis E virus in pigs, a pair of primers were designed according to the ORF2 of swine hepatitis E virus DQ1 strain, and the ORF2 about 369 bp was amplified by PCR. Then the fragment was inserted into pET-32a to establish a expression vector which was named pET32a-DQ1. The sequence analysis showed that the sequence was correct. Then the pET32a-DQ1 was transformed into E.coli BL21, the segment was expressed. The result of SDS-PAGE showed that the expressed protein was about 33ku. Western-blot results indicated that the protein could reacted with swine HEV antiserum. With the protein, a ELISA method for detection of HEV was developed and the primary test revealed the feasibility for clinical use.

Development of Mutiplex PCR for Simultaneous Detection of Staphycococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae and Yeasts in Bovine Mastitis Milk Samples

2006, 37(11): 1202-1208. doi:

Abstract ( 723 )

PDF (1220KB) ( 1046 )

Related Articles | Metrics

A mutiplex polymerase chain reaction(PCR) assay was developed for the simultaneous detection of the four major agents of bovine mastitis, Staphycococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae and yeast. The target sequences of Staphycococcus aureus, Streptococcus agalactiae,and Streptococcus dysgalactiae were the 16S to 23S rRNA spacer regions. The target sequence of yeast was 18S rRNA region. The results showed that the assay was specific. Simulation bacteria infection samples were made according to Skladny’s method. The sensitivity of the mutiplex PCR was lower in detecting Staphycococcus aureus, Streptococcus dysgalactiae and yeast of the milk. The sensitivity of mutiplex PCR in detecting Staphycococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae and yeasts was 104CFU/mL，102CFU/mL，103CFU/mL and 103CFU/mL respectively.The performance of the mutiplex PCR and traditional culture method were evaluated by examining 46 clinical mastitis milk samples and 167 subclinical mastitis milk samples. The results showed that mutiplex PCR was more sensitive than culture in detecting Staphycococcus aureus and yeasts(P<0.01), but was not different from the culture method in detecting Streptococcus agalactiae and Streptococcus dysgalactiae (P>0.05).

The Biological Characteristics of Granulosa Cells Apoptosis of Antritic Follicle in Buffalo Ovary

2006, 37(11): 1209-1216. doi:

Abstract ( 807 )

PDF (1620KB) ( 707 )

Related Articles | Metrics

In ordor to reveal the origination of antral follicular atresia, and to understand if the apoptosis of granulosa cells (GCs) of antrum follicle exists in buffalo ovarian, the buffaloes before puberty and adult buffaloes(at oestrus & dioestrus) were chosen in this study, the apoptotic characteristics of follicular GCs was studied by paraffin section, HE staining, light microscope, DNA in situ end labeling (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL) and transmit electronic microscope(TEM) technology. The results showed that the healthy antrum follicle had not or only had infrequent apoptotic GCs, while atretic antrum follicle had plentiful apoptotic GCs in buffalo before puberty, the representative forms were apoptosis of single or more GCs in GCs layer, and flock of apoptotic bodies in follicular cavity. There were many apoptotic GCs and the GCs layer was curving and crinkling in the big atretic follicle of adult buffalo at dioestrus. GCs apoptosis also exists in Graafian follicle of buffalo at estrus, all GCs in GCs layer fallen into follicular cavity in the big follicle before ovulation. A apoptosis-examining method of the same tissue slice being respectively stained by HE staining and TUNEL has been invented, GCs apoptosis existed in buffalo antral follicles are confirmed by the above method. The GCs nuclear condensed and tended marginization in morphology, as well as apoptotic bodies was oberserved by TEM. Above results suggested that the GCs apoptosis of atretic antral follicles exists in buffalo and the GCs apoptosis are the potential mechanism of arosing antral follicles atresia in buffalo.

Excess Salt Accelerated the Expression of c-jun mRNA in Pulmonary Arterioles Wall in Broiler Chickens

2006, 37(11): 1217-1221. doi:

Abstract ( 1164 )

PDF (1226KB) ( 614 )

Related Articles | Metrics

In this study, the expression of c-jun mRNA in the wall of pulmonary arterioles was investigated to clarify its possible effect during the development of AS induced by excess salt in broiler chickens. One hundred 8 days old male Arbor Acres broilers were randomly divided into control group(C) and treated group (T). Birds in the treated group were supplied with drinking water added 0.30% NaCl, and other factors were same as that in control group. Lung samples of 6 randomly selected birds per group were collected at 15, 22,29,36,43 and 50 days old respectively to make routine paraffin section. Expression of c-jun mRNA was determined by in situ hybridization(ISH) and image analysis program. The results showed that there was dynamic expression of c-jun mRNA in the wall of pulmonary arterioles of broiler chickens. c-jun mRNA was expressed mostly at 22 days old in the control group. The relative content of c-jun mRNA in the wall of all pulmonary arterioles in the treated group was significantly higher than that of control group at 15, 29 and 36 days old (P<0.01), but at 43 and 50 days old significant difference were found only in outer diameter 20-50 μm pulmonary arterioles wall (P<0.01). These results suggested that excess salt accelerated expression of c-jun mRNA particularly in outer diameter 20-50 μm pulmonary arterioles wall，and that expression of c-jun mRNA may play an important role during the development of AS.

Changes of Proliferative Responses and Interleukin-2 Activity of Lymphocytes in Immune Organs of Chickens Primary Infected with E. Necatrix

2006, 37(11): 1222-1225. doi:

Abstract ( 713 )

PDF (1188KB) ( 603 )

Related Articles | Metrics

Dynamic changes of percentage of T cell，interleukin-2(IL-2) activity and proliferative response of T and B lymphocytes to concanavalin A(ConA) or phorble myristate acetate(PMA) in immune organs of 14dayold chickens primary infected with E. necatrix were studied by using acid nonspecific α naphthyl acetate esterase(ANAE), cellular culture technique and ［3-(4,5-dimethylthiazol-2-yl)］-2,5-diphenyltetrazolium bromide (MTT) method. The results showed that the IL-2 activity in thymus and spleen of chickens primary infected with E.necatrix was significantly increased compared with that of control, respectively, at day 16-18 and 18-21 post primary infection (PPI); The proliferative responses of T lymphocytes to ConA in thymus and spleen of chickens infected with E. necatrix were significantly elevated compared with that of control chickens at day 14-16 and 10-18 PPI, respectively. The proliferative responses of B lymphocytes to PMA in spleen and bursa of Fabricius of chickens infected with E. necatrix were significantly higher than that of control at 14-21 and 14-24 PPI, respectively. These results showed that IL-2 immune regulation and function of cellular immune and fluid immune in immune organs of chickens primary infected with E. necatrix had significantly increased.

The Identification of Separated and Cultured Rabbit Endometrial Cellsand the Effect of Sex Hormone on Shape of Endometrial Stromal Cells

2006, 37(11): 1226-1231. doi:

Abstract ( 695 )

PDF (1397KB) ( 594 )

Related Articles | Metrics

To study the morphological characteristics, the secretory capability of endometrial cells and the effect of sex hormone to endometrial cells, different digestion methods and isolation methods were used to digest and isolate endometrial cells from early pregnant rabbit. The results showed that endometrial cells include two types of cells, stromal cells and epithelial cells. Two types of cells were positive respectively with vimentin or cytokeratin by immunocytochemistry. When estradiol and/or progesterone were interfused in the medium, morphological characteristics of stromal cells changed, representing decidualization to some extent.

Effect of Biotin on mRNA Express of PCNA Gene in Speen Cell for Broilers

2006, 37(11): 1232-1235. doi:

Abstract ( 739 )

PDF (1196KB) ( 550 )

Related Articles | Metrics

Molecular biological method was applied to explore the effect of biotin on mRNA expression of PCNA gene in spleen, aimed to probe the mechanism of biotin enhancing spleen cell propagation. 60 AA male broilers of 1 day were allocated into 3 treatments, 20 broilers per treatment. The basal diet was wheat-caseinfish meal diet. Biotin was added in the basal diet at 0.3 mg/kg and 2.7 mg/kg. There were 3 diets including the control of basal diet. In order to stimulate mRNA expression of PCNA gene, The avian flu vaccine was injected quantificationally and hypodermically into broilers on day 48. The results showed that addition of biotin elevated significantly mRNA expression level of PCNA gene in spleen (P<0.01), The mRNA expression level of 2.7 mg/kg treatment was higher significantly than that of the control and 0.3 mg/kg treatment (P<0.01), and no significant difference was founded between 0.3 mg/kg treatments and the control group.(P>0.05). It was concluded that biotin can elevate mRNA expression level of PCNA gene, thus enhance the propagation of spleen cell.

Effect of Malate at Different Concentration on Rumen Fermentation of Goats

2006, 37(11): 1236-1240. doi:

Abstract ( 1046 )

PDF (1237KB) ( 617 )

Related Articles | Metrics

Four goats with permanent rumen fistulae in 4×4 Latin square design were used to study the effect of malate (0, 5, 10, 15 g/d) on rumen fermentation of goats. The results showed that the pH and propionate percentage increased significantly in rumen (P<0.05), and the NH₃-N, lactate concentration and acetate to propionate ratio (A/P) decreased significantly in rumen (P<0.05). The lactate concentration in 5 g/d and 10 g/d malate groups decreased respectively by 31% (P<0.05) and 29% (P<0.05) at 8 h compared with the control. Both of 10 g/d and 15 g/d malate groups decreased the NH₃-N concentration by 30% (P<0.05) at 2 h compared with the control. Compared with the control, 10 g/d and 15 g/d malate groups decreased respectively average daily acetate to propionate ratio by 12% (P<0.05) and 22% (P<0.01). Average daily total volatile fatty acid (TVFA) concentration of all malate groups had no significant change compared with the control. The results suggested that based diets supplemented with malate could improve rumen fermentation type and increase the use of lactate and NH₃-N.

Isolation and Priliminary Identification of Turkey Coronavirus

2006, 37(11): 1241-1244. doi:

Abstract ( 732 )

PDF (1218KB) ( 565 )

Related Articles | Metrics

An acutely infectious disease was encountered in turkey poults of 15-28 days old at a turkey farm in 2003. The affected birds presented with diarrhoea, the hyperaemia and hemorrhage of duodenum and rectum, cecum tumescence, intestines filled with yellow contents and 10% to 20% mortality. Livers, spleens and intestines obtained from died poults were homogenized and centrifuged. The supernatant was inoculated into 15-day-old specific pathogen free (SPF) embryonated chicken eggs to propagate the viruses. After 5 successive passages, the yolk and intestine of infected chicken embryos were harvested and homogenized 72 hours post-inoculation, for determining viral hemagglutination and purifying the viruses. The isolate had the ability to hemagglutinate erythrocytes from rabbit but not from chicken, and several spherical or oval viral particles with corona were observed by electron microscopy, which indicated the isolated viruses may be turkey coronaviruses. Furthermore, TCV S2 gene was amplified from the extracted total RNA of infected chicken embryos intestines by reverse transcriptionpolymerase chain reaction (RTPCR) with designed primers, and the products were identical with the expectation. In addition, 3 days after 1-day-old turkey poults were orally inoculated with 1 mL of the intestine homogenate of the infected chicken embryos, the infected birds showed the same clinical signs and pathological changes as natural infected cases described above. Above results showed that the isolated viruses were turkey coronaviruses. It is the first time that coronaviruses were isolated from turkey in China.

Table of Content