ABSTRACT
Microtus fortis (reed vole) is the only mammal known to have natural resistance to Schistosomiasis japonica. Originating from schistosomiasis endemic and non-endemic areas, as well as laboratory bred voles have the same resistance to Schistosoma japonicum. After more than 30 years of laboratory cultivation of wild reed vole, a series of progress have been made in laboratory animalization. A detailed study was conducted on biological traits including growth and development, reproductive physiology, serum biochemistry, hematological indicators and tissue anatomy. At the same time, the anti-schistosomiasis characteristics and anti-schistosomiasis mechanisms of Microtus fortis were studied. The closed Dongtinghu population of Microtus fortis (S: DTMF) cultivated by Shanghai Laboratory Animal Research Center was recognized as a Chinese laboratory animal resource by the Experimental Animal Resources and Evaluation Working Committee of the Chinese Association for Laboratory Animal Sciences in 2021. This review focuses on summarizing the research progress in the biological characteristics, standardization research, genome and anti-schistosomiasis mechanism of reed vole in the past decade, especially in the implementation of the key project in the National Science and Technology Pillar Program.
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ObjectiveTo establish a method for rapid and sensitive detection of Staphylococcus aureus. MethodsThe specific gene nuc of Staphylococcus aureus was selected as the target gene. A pair of specific primers and a TaqMan probe were designed and synthesized according to the published sequence of the nuc gene. Establish a nucleic acid detection method for nuc gene using fluorescence quantitative PCR technology, and apply it clinically in the detection of fecal samples from rats and mice. ResultsThe DNA extracted from Staphylococcus aureus and other non-Staphylococcus aureus strains was detected by qPCR. The results showed that Staphylococcus aureus had a specific amplification curve, while other non-Staphylococcus aureus did not, indicating that the designed primers and probes were specific for Staphylococcus aureus. The sensitivity of this method was determined by diluting the DNA of Staphylococcus aureus by 10 times. The results showed that the detection limit of this method was 10 fg DNA, which was 2 orders of magnitude higher than that of ordinary PCR method. A total of 91 clinical samples were detected in this study, of which 4 rat samples from the same facility had a typical S-curve. The PCR products were sequenced and BLAST compared. The gene sequence of this sample was 100% similar to that of Staphylococcus aureus, indicating that the sample was positive for the nucleic acid of Staphylococcus aureus nuc gene, with a positive rate of 4.40%. The result was consistent with that obtained by bacterial culture method. The nucleic acid extraction adopted a full-automatic nucleic acid purification instrument, and the time required from nucleic acid extraction to detection result determination was less than 1.5 h. ConclusionThe qPCR method established in this study to identify Staphylococcus aureus with nuc gene as the target gene has the advantages of fast, high sensitivity and specificity, and can be used for the detection of Staphylococcus aureus in feces of rats and mice.
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Objective To compare the differences of bacterial distribution of intestinal flora in Microtus fortis living under laboratory feeding and wild survival conditions. Methods The 16S rDNA-V4-V5 region of bacteria in the ileocecal contents from Microtus fortis raised in lab and captured in wild were measured by high-throughput sequencing. The number of operational taxonomic units(OTUs)were sorted and calculated,and the species abundance and distribution and difference were analyzed. Results The rarefaction curves indicated that adequate sampling was achieved. At the phylum level,the distribution of intestinal flora between two groups was similar. The experimental group had a unique phylum, Lentisphaerae. The wild type group had 3 unique phylums,Fusobacteria,Thaumarchaeota and an unclassified phylum. At the genus level, the kind of intestinal flora in the wild type group was more abundant than the experimental group. Ruminococcus is the largest differential genus. Conclusions The microbial community structure and differences of Microtus fortis living under different conditions are obtained. It may further enrich the basic biology data of Microtus fortis.
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Objective To more intuitively understand the quality control for laboratory animals and further achie-ving a more scientific and reasonable management of laboratory animals, the infection index as evaluation criteria was intro-duced. Then the best way to calculate infection index was explored in order to more scientifically reflect the infection status of laboratory animals. Methods Infection index, also called the degree of infection, is a qualitative indicator of monito-ring laboratory animal quality. After arranging, analyzing, processing and gathering the data from laboratory animal quality monitoring, the index reflects synthetically the pathogen infection status or trend of a particularly investigated experimental animal population or the development of certain experimental animals. Results In general, the pathogen infection index of mice was slightly decreased, while the pathogen infection index of rats roughly increased year by year. In comparing infec-tion index by different pathogens, the parasite infection index of mice was found to be higher than bacteria and virus infec-tion indexes, while the bacteria infection index of rats was higher than parasite infection index and virus ones. Conclusions The infection index model intuitively reflects the quality control status of laboratory animals. The analysis also reveals that the parasite monitoring of the mice and the bacteria detection of rat needs to be reinforcement. In addition, the index of infection reveals that the pathogen infection of mice is well under control, while that of rats tends to be more serious year by year.
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Objective To determine the interference effect of H. hepaticus infection on the functional characteris-tics of dendritic cell ( DC) surface molecules and immune response in mice. Methods Male BALB/c mice were inocula-ted with H. hepaticus (ATCC 51450). Murine bone marrow-derived dendritic cells (DC) were isolated and co-cultured which were stimulated by GM-CSF and IL-4 at the fifth month after the last inoculation. Then the DCs were subjected to FACS analysis for surface markers (CD11c, CD40, CD80 and MHCII) detection. On this basis, virus suspension of New-castle disease virus( NDV) ZJ1 strain was inoculated into the mice. Serum was collected for detection of the NDV antibody titer in serum weekly to explore the difference of antibody titer between the two groups. Results The expression rates of CD40 and MHCII on the mouse DCs in experimental group were higher than that in the control group. The NDV antibody ti-ter of experimental group was slightly lower than that in the control group in the first week. During the 2nd to 5th weeks, the titer was higher than that in the control group, with a very significant difference. In the 6th week, the titer of both the two groups tended to fall. Conclusions H. hepaticus infection can promote bone marrow DC maturation in mice, stimulate the expression rates of MHC II and CD40, and enhance the NDV antibody levels.
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Objective To compare the efficiency of bacteria culture and PCR assays for detection of Staphylococcus aureus ( S.aureus) , Pseudomonas aeruginosa ( P.aeruginosa) and Klebsiella pneumoniae ( K.pneumoniae) in laboratory rats and mice.Methods Bacteria culture combined with biochemical identification and PCR assay were used to detect 78 SPF rats and 422 SPF mice and the results of the two methods were compared .Results All the 78 rats were negative .Of the 422 mice, the positive rate by culture was 7.11%(30/422), of which, 10 were S.aureus, 22 were P.aeruginosa, and 2 were K.pneumoniae.The positive rate by PCR was 7.58%(32/422), of which, 10 were S.aureus, 25 were P. aeruginosa, and 2 were K.pneumoniae.Conclusions The high sensitivity , rapid procedure and easy to operate of PCR assay makes it valuable for rapid bacteria diagnosis and large-scale screening in laboratory animals .
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Objective To obtain the full-length cDNA sequences of CYP2E1,CYP2D5,ECHS1,which may be related with non-alcoholic fatty liver disease,from Microtus fortis.Methods To construct Microtus fortis liver cDNA plasmid library using SMART technique,to get the purposed colonies through screening libraries by PCR,and to obtain their full-length cDNA sequences by sequencing with pBluescript II SK universal primers M13R.Results Three full-length cDNA sequences of Microtus fortis,CYP2E1,CYP2D5 and ECHS1 were obtained.The CYP2E1 cDNA was 1685 bp in length and contained a 1482 bp open reading frame(ORF) encoding a 494 amino acids.The CYP2D5 cDNA was 1690 bp in length,and contained a 1514 bp ORF encoding 504 amino acids.The ECHS1 cDNA was 1013 bp in length,and containsed an 873 bp ORF encoding 290 amino acids.Sequence analysis revealed that the identity of the three cDNA sequences and deduced amino acids among Microtus fortis,Homo sapiens,Mus musculus and Rattus norvegicus was high.Conclusion The full-length cDNA sequences of CYP2E1,CYP2D5,ECHS1 were obtained from Microtus forti,liver cDNA library.and the gene sequences have been deposited in GenBank (GQ507485,GQ507486,GQ845171),which may lay the foundation for researchies of pathogenesis of non-alcoholic fatty liver disease in Microtus fortis models.