Identification and characterization of duck plague virus glycoprotein C gene and gene product.

BACKGROUND: Viral envelope proteins have been proposed to play significant roles in the process of viral infection. RESULTS: In this study, an envelope protein gene, gC (NCBI GenBank accession no. EU076811), was expressed and characterized from duck plague virus (DPV), a member of the family herpesviridae. The gene encodes a protein of 432 amino acids with a predicted molecular mass of 45 kDa. Sequence comparisons, multiple alignments and phylogenetic analysis showed that DPV gC has several features common to other identified herpesvirus gC, and was genetically close to the gallid herpervirus.Antibodies raised in rabbits against the pET32a-gC recombinant protein expressed in Escherichia coli BL21 (DE3) recognized a 45-KDa DPV-specific protein from infected duck embryo fibroblast (DEF) cells. Transcriptional and expression analysis, using real-time fluorescent quantitative PCR (FQ-PCR) and Western blot detection, revealed that the transcripts encoding DPV gC and the protein itself appeared late during infection of DEF cells. Immunofluorescence localization further demonstrated that the gC protein exhibited substantial cytoplasm fluorescence in DPV-infected DEF cells. CONCLUSIONS: In this work, the DPV gC protein was successfully expressed in a prokaryotic expression system, and we presented the basic properties of the DPV gC product for the first time. These properties of the gC protein provided a prerequisite for further functional analysis of this gene.

Isolation, identification of Streptococcus pneumoniae from infected rhesus monkeys and control efficacy.

BACKGROUND: Streptococcus pneumoniae can cause a wide variety of illnesses. Primate animals can be infected by the pneumococcus. A disease occurred among rhesus monkeys in winter 2006. METHODS: Routine clinical observation, necropsies, bacteriological examinations were conducted, and PCR, pathogenicity to BALB/c mice and antibiotic susceptibility test were examined additionally. RESULTS: We conclude that the agent is S. pneumoniae. Based on the antibiotic susceptibility test, a dose of 20 mg/kg body weight daily of Erythromycin was given intramuscular injection for 5 days, resulting in the disappearance of clinical signs, and no newly case reappear be observed till today. CONCLUSIONS: Therefore, it is suggested that the outbreak of respiratory disease in the rhesus monkeys was because of transmission of S. pneumoniae among rhesus monkeys. The antibiotic therapy finding underscores the utility of Erythromycin to cure the infected rhesus monkeys without causing side effects and without contributing to the further development of antibiotic resistance.

[Polymorphism of MHC-DPB1 gene exon 2 in rhesus macaques (Macaca mulatta)].

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.

In vivo assessment of mitochondrial toxicity of metacavir in Rhesus monkeys after three months of intravenous administration.

AIM: To explore the potential mitochondrial toxicities and their severities of intravenously administered metacavir, a nucleoside analog, in rhesus monkeys. METHODS: 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. RESULTS: 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. CONCLUSION: These results suggest that metacavir has not a high risk for potential mitochondrial-related effects in rhesus monkeys.

A case report: Entamoeba histolytica infections in the rhesus macaque, China.

A case of amebic dysentery due to a natural infection of Entamoeba histolytica in the rhesus macaque (Macaca mullata) was reported. A fecal specimen was isolated and identified by the polymerase chain reaction technique. A daily dose of 750 mg of metronidazole was given orally for 10 days, and good results were observed. The early diagnosis of an E. histolytica infection allowed a proper antiamoebic treatment in an early stage of infection resulting in a successful outcome after therapy.

Comparison of efficacy of selamectin, ivermectin and mebendazole for the control of gastrointestinal nematodes in rhesus macaques, China.

An experiment was conducted to evaluate the efficacy of ivermectin and mebendazole compared with selamectin against gastrointestinal nematodes in rhesus macaques. A total of 60 rhesus macaques (Macaca mulatta), which were all infected with gastrointestinal nematodes, were randomly assigned to three treatment groups (selamectin, ivermectin and mebendazole) and one control group. Fecal samples for determining nematode egg counts were collected pre- and post-treatment. All treatments resulted in decrease in the number of eggs per gram (EPG) in the post-treatment sample compared with the pre-treatment sample. Reductions of mean egg counts from day -3 levels were 99.4% for selamectin, 99.2% for ivermectin and 99.4% for mebendazole on trial day 11, respectively. However, no significant difference was found among treatment groups. According to the data demonstrating a similar efficacy in selamectin-, ivermectin- and mebendazole-treated rhesus macaques, it was effective and convenient to apply either selamectin and ivermectin or mebendazole in rotation on the local conditions.