-
[show abstract]
[hide abstract]
ABSTRACT: Because wild rhesus macaque (Macaca mulatta) populations have suffered major declines, there is a growing need to characterize their genetic and population structure in order to protect the genetic integrity of this species. In this study, we genotyped a sample comprising 120 wild rhesus macaques from six sites in Sichuan Province for 30 nuclear microsatellite (STR) loci using an ABI 3130xl genetic analyzer. Bayesian analyses and PCA clearly differentiated monkeys from Heishui from those at other sites. The samples from all six sites exhibited high gene diversity suggesting that the Sichuan wild rhesus macaque populations are not threatened by a lack of genetic diversity. Deviation from Hardy-Weinberg equilibrium was more frequent in the Danba and Heishui populations. This may be due to the more fragmented habitat and less disturbance by humans in this area that foster greater subpopulation structuring than occurs in eastern China. We suggest that this population subdivision is the result of both long-term geographic barriers and human activity.
Molecular Biology Reports 12/2012; · 2.93 Impact Factor
-
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 09/2011; 27(5):501-4.
-
[show abstract]
[hide abstract]
ABSTRACT: Between one and six subspecies of Chinese rhesus macaques (Macaca mulatta) have been proposed based on morphological differences and/or their geographic distribution. In this study, a 489 base pair fragment of the mitochondrial control region was amplified from 230 DNA samples collected from rhesus macaques in the Sichuan province in Western China. The fragment was then sequenced and aligned with 208 sequences from wild rhesus macaques, sampled throughout the species' geographic range in China downloaded from GenBank. Phylogenetic analysis of the 182 unique sequences identified among these samples divided Chinese rhesus macaques into two western haplogroups (haplogroups A and B) and three older eastern haplogroups (haplogroups C, D, and E), whose differentiation probably occurred during the penultimate glacial event. During the warming after the penultimate glacial event, haplogroups A, B, and E rapidly expanded and a relatively young subhaplogroup of haplogroup E, E', limited to Southern China but shared with Vietnamese rhesus macaques, was reintroduced from Indochina during the last glacial event. One haplotype most closely related to subhaplogroup E' probably represents the isolation of Hainan Island, to where it is restricted, from the mainland by the formation of the Qiongzhou Strait approximately 8,500 years ago. The distribution of haplogroups both informs the phylogeographic history of dispersal of Chinese rhesus macaques and has implications for their suitability as animal models in biomedical research.
American Journal of Primatology 04/2011; 73(9):883-95. · 2.22 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The herpesvirus UL34 gene is highly conserved in the Herpesviridae family. The nucleotide sequence has the characteristic of comparatively high content of G + C and the gene product is a tail-anchored type II membrane protein involved in the primary envelopment of capsid in the endoplasmic reticulum, profiting the virion maturation and nucleocapsid budding, regulating the proper location of the UL31 protein and lamin in infected cells. In this work, we summarize the research progress on characteristic and function of the UL34 gene and its encoding protein to provide some new insights into the molecular biology of herpesvirus and its mechanism of replication.
Reviews in Medical Microbiology 03/2011; 22(2):22–27. · 0.37 Impact Factor
-
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 09/2010; 26(5):414-7.
-
Tao Zhou,
Dekang Zhu,
Xiaoyue Chen, An-chun Cheng,
Ming-shu Wang,
Renyong Jia,
Qihui Luo,
Hengmin Cui,
Yi Zhou,
Yin Wang,
Zhiwen Xu,
Xiaoyu Wang
[show abstract]
[hide abstract]
ABSTRACT: We determined the nucleotide sequence of a portion of the duck enteritis virus (DEV) strain CHv which was suspected to be the UL10 gene(GenBank accession no EU195100.), encoding glycoprotein M(gM). The UL10 gene was cloned and sequenced, which was composed of 1230 nucleotides encoding 409 amino acids, with a predicted molecular mass of 45.752 kDa. We calculated the codon usage bias in the newly identified UL10 gene of Duck Enteritis Virus (DEV) and performed a correlation analysis of codon bias of DEV UL10 gene. There is a strong bias in the DEV UL10 gene towards the synonymous codons with A and T at the third codon position. Multiple sequence alignment suggested that the UL10 gene was highly conserved in Alphaherpesvirinae and similar to the other herpesviral UL10. Phylogenetic analysis showed that the gene had a close evolutionary relationship with the avian herperviruses, and DEV should be placed into a single cluster within the subfamily Alphaherpesvirinae. Furthermore, the UL10Δ gene was cloned into a pET prokaryotic expression vector and transformed into Escherichia coli BL21 (DE3). A 6 kDa fusion protein was induced by the further culture at 30°C after addition of 0.8mM IPTG. Our results may provide some insight for further research about the gene and also enrich the database of herpesvirus.
Bioinformatics and Biomedical Engineering (iCBBE), 2010 4th International Conference on; 07/2010
-
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 07/2010; 26(4):340-4.
-
[show abstract]
[hide abstract]
ABSTRACT: Rhesus macaque (Macaca mulatta) has long been used as an experimental model animal for biomedical research and was under the key state protection (class II) from Chinese government. In order to facilitate the use of Chinese rhesus macaques in biomedical research and their protection based on better understanding of the major mistocompability complex (MHC) genes in these macaques, the exon 2 of Mamu-DPB1 genes were determined in 106 wild rhesus macaques using DGGE, cloning and sequencing. A total of 21 Mamu-DPB1 alleles were obtained, of which 15 alleles were novel sequences that had not been documented previously. Mamu-DPB1 30 was the most frequent allele in the whole large population comprising all 106 rhesus macaque individuals (0.1120) and in Xiaojin population (0.1120), Mamu-DPB1 04 in Heishui (0.1702), -DPB1 32 in Bazhong (0.1613), -DPB1 30 in Hanyuan (0.1120), and -DPB1 04 in Jiulong (0.1139). The alignment of the amino acids sequences showed that 12 variable sites were species-specific, of which 9 sites occurred in the putative amino acids sequences of the 15 novel Mamu-DPB1 alleles. Trans-species polymorphism was observed on the phylogenetic tree based on the DPB1 alleles of rhesus macaques and cynomolgus (Macaca fascicularis). In addition, these results also demonstrated that significant genetic differentiation has occurred between Chinese and Indian rhesus macaque population.
Hereditas (Beijing) 06/2010; 32(6):588-98.
-
[show abstract]
[hide abstract]
ABSTRACT: Based on the duck plague virus (DPV) UL35 gene sequence that our laboratory obtained (GenBank accession number EF643558), a pair of primers was designed using Oligo6.0 and primer5.0, then the UL35 gene was amplified from DPV CHv strain genomic DNA and cloned into the pMD18-T to construct a clone plasmid pMD18-T-UL35. After identification of the pMD18-T-UL35 by PCR amplification and restriction digestion, the fragment of the UL35 gene was subcloned into the prokaryotic expression vector pET-32a(+). The resultant recombinant plasmid pET-32a(+)-UL35 was then transformed into E. coli BL21 (DE3) strain and optimally-expressed under the induction of 1.0 mmol/L IPTG at 34 degrees C for 5 hours. SDS-PAGE analysis showed the recombinant protein (VP26) had a molecular weight of about 33KDa and accounted for 32.3% of total bacterial protein by gel scanning. The protein was then purified by Ni(2+)-affinity chromatography and used to immunize rabbit for producing the VP26 anti-serum and its antibody titer was up to 1:32 detected by agar diffusion reaction. After the IgG of the polyclonal antibodies was purified by High-Q anion-exchange chromatography, Western blot analysis indicated that the IgG had specific reaction with the VP26. Moreover, the subcellular localization detection was observed using immunofluorescence technique. The results showed that the specific fluorescences appeared relatively few in nucleus in 2 to 8 hours and increased gradually in 12 to 36 hours and eventually reached to the maximum, which aggregated in the spot region of the nucleus after the duck embryo fibroblast (DEF) were infected by DPV. However, there were only a small amount of specific fluorescences in the cytoplasm in 12 hours and increased with the extension of infection time in 24 to 48 hours. The specific fluorescences finally reached to the maximum in the cytoplasm in 72 hours. The results provided significant data for furthering the study on the function of DPV UL35 gene.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 03/2010; 26(2):143-9.
-
[show abstract]
[hide abstract]
ABSTRACT: To explore the toxicities of metacavir, a novel deoxyguanosine analog with an anti-hepatitis B virus (HBV) potential, in a 6-month repeated dosing in rhesus monkeys.
Rhesus monkeys were divided into four groups with eight animals in each group. Metacavir or blank vehicles were given for up to 6-month, and then the animals were euthanized 3 and 6 months later. Biochemical and hematological parameters, general symptoms, ECG, serum antibodies, and tissue pathology were observed and recorded.
No biologically meaningful influences on body weight, body temperature, ocular examination, ECG, and organ weight were observed. The main toxic effects included: obvious gastrointestinal toxicities were observed in metacavir 200 mg/kg group, in which animals experienced vomiting and decrease in food consumption. Along with the increase of dosing times, animals gradually tolerated the drug and all these effects gradually abated. Hematological damages included increased damage of red blood cells, decrease of red blood cell count and hemoglobin levels. Hepatic functions were also damaged at certain levels, including the decreases in total protein, albumin, and glucose and the fatty degeneration in hepatocytes. Mild stenosis and exfoliation of gastric and duodenal mucosa was observed. The mild necrosis and exfoliation of renal tubules epithelia was found 6 months after the start of dosing. All these toxic effects were dose dependent.
The main target organs of the toxic effects of metacavir are gastrointestinal tract, liver, blood, and kidneys. The no-observed-adverse-effect-level (NOAEL) of metacavir for rhesus monkey were considered to be 50 mg/kg/day.
Experimental and toxicologic pathology: official journal of the Gesellschaft fur Toxikologische Pathologie 03/2010; 63(4):379-85. · 1.43 Impact Factor
-
Ai-Mei Shen,
Guang-Peng Ma, An-Chun Cheng,
Ming-Shu Wang,
Dan-Dan Luo,
Li-Ting Lu,
Tao Zhou,
De-Kang Zhu,
Qi-Hui Luo,
Ren-Yong Jia,
Zheng-Li Chen,
Yi Zhou,
Xiao-Yue Chen
[show abstract]
[hide abstract]
ABSTRACT: Some UL45 gene function of Herpesvirus was reported. While there was no any report of the duck enteritis virus (DEV) UL45 protein as yet.
The UL45 gene and des-transmembrane domain of UL45 (named UL45Δ gene, 295-675bp of UL45) of DEV were amplified by PCR and subcloned into the prokaryotic expression vector pET-32a(+). The constructed recombinant plasmids were transformed into the host strain BL21(DE3) PLysS and induced by IPTG. SDS-PAGE analysis showed the UL45 gene couldn't express while UL45Δ gene was highly expressed. His Purify Kit or salting-out could purify the protein effectively. Using the purified protein to immunize New-Zealand rabbits and produce polyclonal antibody. The agar diffusion reaction showed the titer of antibody was 1:32. Western blot analysis indicated the purified rabbit anti-UL45Δ IgG had a high level of specificity and the UL45 gene was a part of DEV genome. The transcription phase study of UL45 gene showed that expression of UL45 mRNA was at a low level from 0 to 18 h post-infection (pi), then accumulated quickly at 24 h pi and peaked at 42 h pi. It can be detected till 72 h pi. Besides, western blot analysis of purified virion and different viral ingredients showed that the UL45 protein resided in the purified virion and the viral envelope.
The rabbit anti-UL45Δ IgG was produced successfully and it can serve as a good tool for penetrating studies of the function of DEV UL45 protein. The transcription phase and protein characteristics analysis indicated that DEV UL45 gene was a late gene and UL45 protein may be a viral envelope protein.
Virology Journal 01/2010; 7:232. · 2.34 Impact Factor
-
Ming-Sheng Cai, An-Chun Cheng,
Ming-Shu Wang,
Wan-Ping Chen,
Xian Zhang,
Shang-Xi Zheng,
Yang Pu,
Kun-Peng Lou,
Yao Zhang,
Lei Sun,
Lu-Lu Wang,
De-Kang Zhu,
Qi-Hui Luo,
Xiao-Yue Chen
[show abstract]
[hide abstract]
ABSTRACT: Previous study has demonstrated that the duck plague virus (DPV) UL35 gene can be expressed as a recombinant fusion protein, and the prepared antiserum has a high reactivity and specificity against the purified recombinant protein. In the present study, to elucidate the properties and functions of its encoding protein, the UL35 gene product (VP26) was identified by using the prepared rabbit polyclonal antiserum.
Real-time PCR, Western blot and immunofluorescence analysis were used to determine the transcription and expression kinetics and subcellular localization of DPV VP26 in DPV-infected cells.
A protein of approximately 13 kDa that reacted with the antiserum was detected in immunoblot of DPV-infected cellular lysates. Real-time PCR and Western blot analysis of DPV-infected cells showed that VP26 was produced predominantly at the late stage of infection, its production was highly dependent on viral DNA synthesis, and the UL35 gene was regulated as a late viral gene, suggesting that the gene should be categorized as gamma2 class. Additionally, analysis of the association of DPV VP26 with purified virions revealed that VP26 was a component of extracellular mature DPV virions. Subcellular localization demonstrated that VP26 firstly localized in cytoplasm, then it transferred to the nucleus and aggregated in the punctate region of the nucleus in DPV-infected cells.
Taken together, these results will provide a foundation for further functional analysis of the DPV UL35 gene.
Intervirology 01/2010; 53(6):408-16. · 2.34 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Kirby-Bauer tests were used to analyze the antibiotic resistance of 224 isolates of Riemerella anatipestifer isolated between 1998 and 2005. Among the 36 antibiotics tested, 50% of the analyzed isolates were resistant to ampicillin, ceftazidime, aztreonam, cefazolin, cefepime, cefuroxime, oxacillin, penicillin G, rifampin, and trimethoprim/sulfamethoxazole. Higher levels of resistance were detected for aztreonam, cefepime, oxacillin, penicillin G, ceftazidime, and trimethoprim/sulfamethoxazole (87.8%, 64.3%, 88.6%, 86.9%, 75.9%, and 79.2% resistance, respectively). The lowest resistance rates were observed for amikacin (9.5%), cefoperazone (7.2%), imipenem (3.2%), and neomycin (9.5%). Four isolates were found to be resistant to 29 different antimicrobials. Riemerella anatipestifer drug resistance profiles changed over time, and the only consistent patterns observed were the resistance of R. anatipestifer to cefoperazone, piperacillin, spectinomycin, and aztreonam. In addition to determining the antibiotic-resistance profiles of R. anatipestifer isolates, we also examine the therapeutic efficacy of these antibiotics against lethal R. anatipestifer infection in ducks in vivo. According to these data, we have extrapolated an antibiotic treatment approach for veterinarians attending flocks of ducks. These data suggest that disk-diffusion analyses can be extrapolated to predict in vivo efficacy, thereby facilitating the identification of effective antibacterial treatments and potentially diminishing the irresponsible use of antibiotics.
Avian Diseases 12/2009; 53(4):601-7. · 1.46 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To explore the potential mitochondrial toxicities and their severities of intravenously administered metacavir, a nucleoside analog, in rhesus monkeys.
Totally 21 rhesus monkeys were randomly divided into 4 groups: metacavir 120 mg/kg group, metacavir 40 mg/kg group, zidovudine(AZT) 50 mg/kg group, and blank control group. Animals were killed after the completion of dosing or further observed in a 4-week recovery phase. Changes of structure of mitochondria in liver, kidney, skeletal muscles, and cardiac muscles were observed under transmission electron microscope(TEM). Changes of the activities of mitochondrial respiratory chain complexes and mitochondrial DNA were also determined.
In metacavir 120 mg/kg group, some mitochondrial injuries were found in skeletal muscle, cardiac muscle, and liver, including that some cristae was broken and became sparse in density in the skeletal muscle, the morphology and size of mitochondria remained unchanged. Metacavir decreased the activities of respiratory chain complexes I and II and the mtDNA contents in three tissues in a dose-dependent manner; however, the extent of such decrease was lower than that in AZT 50 mg/kg group. The mitochondrial injuries in metacavir 40 mg/kg group were mild in each tissue and no obvious change in mitochondrial function was noted. On week 4 in the recovery phase, results showed that all these injuries were reversible after drug withdrawal.
These results suggest that metacavir has not a high risk for potential mitochondrial-related effects in rhesus monkeys.
Acta Pharmacologica Sinica 11/2009; 30(12):1666-73. · 1.95 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Identificación y caracterización de un gene similar al US3 del virus de la enteritis de los patos. El virus de la enteritis de los patos causa pérdidas sustanciales en las granjas de patos, sin embargo, se conoce poco acerca de su biología molecular. En este estudio, se identificó como parte de una librería genética un marco de lectura continuo de un gen similar al US3 del virus de la enteritis de los patos. Su existencia fue confirmada por clonación molecular a partir de fibroblastos de pato infectados con el virus y por la secuenciación de ADN. El gene similar al US3 fue subclonado posteriormente en un sistema procariótico de expresión de proteínas y se expresó en Escherichia coli como una proteína de fusión con un marcador de seis histidinas. La proteína fue purificada e inoculada en conejos para la producción de antisuero. Se generó de manera exitosa un anticuerpo primario específico para el gene y se utilizó para detectar proteínas similares a la US3 en células de patos infectadas con dicho virus. Se demostró la expresión in vivo de una proteína similar a la US3 en fibroblastos de embrión de pato infectados con el virus mediante el ensayo de inmunofluorescencia indirecta y microscopía de fluorescencia regular, mientras que los fibroblastos de pato no infectados no mostraron tinción específica. Además, se utilizaron el ensayo de inmunofluorescencia indirecta y microscopía confocal para estudiar el curso y la localización subcelular de la expresión de la proteína. Se determinó que la proteína se expresa tan temprano como a las dos horas postinfección y su expresión fue incrementada con el tiempo a las cuatro, ocho, doce y 24 horas después de la infección. Se determinó que la proteína se localiza principalmente en el área perinuclear y en el citosol, y posteriormente se detectó en el núcleo. Además, se construyó un árbol filogenético con secuencias de la proteína US3 que demostró que la relación evolutiva del virus de la enteritis de los patos está cercana al género Mardivirus. En concreto, se detectó por primera vez un gene similar al US3 del virus de la enteritis de los patos y se estudio su expresión in vivo, se aportaron sugerencias para la clasificación de dicho virus y en la determinación de las funciones del gene. Abbreviations: CDD = conserved domain database; CeHV = cercopithecine herpesvirus; DEF = duck embryo fibroblast; DEV = duck enteritis virus; EDTA = ethylenediaminetetraacetic acid; HSV = herpes simplex virus; HVP = baboon herpesvirus; IgG = immunoglobulin G; MDV = Marek's disease herpesvirus; MEM = minimum essential medium; NLS = nuclear localization signal; NCBI = National Center for Biotechnology Information; ORF = open reading frame; PBS = phosphate-buffered saline; VZV = human herpesvirus 3
Avian Diseases 10/2009; · 1.46 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The aim was to identify the codon usage bias between the newly identified duck plague virus (DPV) UL35 gene (GenBank accession No. EF643558) and the UL35-like genes of 27 other reference herpesviruses.
A comparative analysis of the codon usage bias of the 28 herpesviruses was performed by using the CodonW 1.4 program and CUSP (create a codon usage table) program of EMBOSS (The European Molecular Biology Open Software Suite).
The results showed obvious differences of the synonymous codon usage bias in the 28 herpesviruses indicated by the Codon Adaptation Index, effective number of codons (ENc), and the value of G + C content at the 3rd codon position. The codon usage pattern of the DPV UL35 gene was phylogenetically conserved and similar to that of the UL35-like genes of the avian alpha-herpesvirus, with a strong bias towards the codons with A and T at the 3rd codon position. A cluster analysis of codon usage pattern of the DPV UL35 gene with other reference herpesviruses demonstrated that the codon usage bias of the UL35 genes of the 28 herpesviruses had a very close relation with their gene function. The ENc-plot revealed that the genetic heterogeneity in the DPV UL35 gene and the 27 reference herpesviruses were constrained by G + C content, while gene length exerted relatively weaker influences. In addition, comparisons of the codon preferences in the UL35 gene of DPV with those of Escherichia coli, yeast and humans revealed that there were 33 codons showing distinct usage differences between the DPV and yeast, and 38 between the DPV and humans, but only 31 between the DPV and E. coli. Therefore, the E. coli system may be more suitable for the expression of the DPV UL35 gene.
Together, these results may improve our understanding of the evolution, pathogenesis and functional studies of DPV, as well as contribute significantly to the area of herpesvirus research and possibly studies with other viruses.
Intervirology 09/2009; 52(5):266-78. · 2.34 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Duck enteritis virus (DEV) causes substantial losses on duck farms; however, its molecular biology is poorly understood. Here, an open reading frame of a US3-like gene of DEV was identified from a DEV genomic library. Its existence was confirmed by cloning from DEV-infected duck embryo fibroblasts (DEFs) and DNA sequencing. The US3-like gene was then subcloned into a prokaryotic protein expression vector and expressed as a six-histidine-tagged fusion protein in Escherichia coli. The protein was purified and inoculated into rabbits for antiserum production. A primary antibody specific to the gene was successfully generated and used to detect the US3-like protein in DEV-infected duck cells. In vivo expression of the US3-like protein in DEV-infected DEFs was demonstrated with indirect immunofluorescence assay and regular fluorescence microscopy, whereas uninfected DEFs did not show any specific fluorescent staining. Furthermore, indirect immunofluorescence assay and confocal microscopy were used to study the time course and subcellular localization of the protein expression. The protein was found to be expressed as early as 2 hr postinfection, and its expression was increased by time at 4, 8, 12, and 24 hr postinfection. The protein was found to be localized mostly around the perinuclear area and in the cytosol, and also in the nucleus at later time points. In addition, a US3 protein phylogenetic tree was constructed and showed that the evolutionary relationship of DEV is close to the genus Mardivirus. In short, the DEV US3-like gene and its in vivo protein expression were found for the first time, and DEV classification and the gene's functions were suggested.
Avian Diseases 09/2009; 53(3):363-9. · 1.46 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Compared to the UL51 gene of other alphaherpesviruses, the duck enteritis virus (DEV) UL51 gene contains ten conserved motifs and has a close evolutionary relationship with members of the genus Mardivirus. The DEV UL51 gene product was identified using a rabbit polyclonal antiserum raised against a 6-His-UL51 fusion protein expressed in Escherichia coli as a 34-kDa protein. Western blotting and RT-(real time) PCR analysis of DEV-infected cells showed that the protein was produced at the late stage of infection and that its production was highly dependent on viral DNA synthesis, suggesting that the gene should be classified as gamma2 class. Analysis of extracellular virions revealed that the protein was a component of extracellular mature DEV virions. Indirect immunofluorescence studies localized most of the protein to the juxtanuclear region. These results will provide a basis for further functional analysis of the gene.
Archives of Virology 07/2009; 154(7):1061-9. · 2.11 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: In order to define clearly the conditions leading to the outcome of acute duck hepatitis B virus (DHBV) infection, 1-day-old Pekin ducklings were infected with DHBV by different routes and given different doses of inoculum. Groups of 24 ducklings were inoculated either intravenously via the vena cruralis, or intraperitoneally with pooled serum containing either 1.6 x 10(7) or 1.6 x 10(4) DHBV genomes. One control duck from each group was inoculated with an equal volume of normal duck serum. A sensitive and reproducible real-time polymerase chain reaction assay based on TaqMan technology was developed for the detection and quantitation of DHBV DNA in the serum and liver. DHBAg was observed in the hepatocytes by immunohistochemistry. Histological changes in the liver tissue were also observed. The results demonstrate that ducklings at each time point and in all groups developed detectable viraemia. In each group, DHBV DNA in the liver was at a lower level than in serum and the peak DNA titre was found in serum earlier than in the liver. In the low-dose groups it was always at a lower level than in the high-dose groups. The DHBV replication levels appeared to be directly related to the number of DHBAg-positive hepatocytes. The variation trends of DHBAg-positive hepatocytes were similar in the high-dose groups. Histological changes were associated with liver viral DNA levels. We suggest that this dose and route of inoculation can be used as a model to study acute DHBV infections.
Avian Pathology 05/2009; 38(2):129-34. · 1.71 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) is a molecular biological technology that can be used to study microbial community diversity and dynamics. In many reports, investigations of microbial diversity from environmental samples were based on the agarose gel electrophoresis (AGE) patterns of ERIC-PCR amplified products. This is not a sound practice, since bands with identical positions can contain different sequences; thus, this practice could possibly exaggerate the similarities or diversities among samples. To mitigate this issue, we employed a denaturing gradient gel electrophoresis (DGGE) strategy to explore DNA bands with the same size, between ERIC-PCR profiles of samples. DPS software was used with Shannon-Wiener diversity index (H') to analyze ERIC-PCR fingerprint profiles. H' revealed that the microbial community diversity at DGGE was higher than with AGE. The results of this study suggest that the ERIC-PCR assays with DGGE can provide a better assessment of electrophoresis pattern with regards to the structure of an intestinal microbial community.
Journal of microbiological methods 02/2009; 77(1):63-6. · 2.43 Impact Factor
