摘要：
In China, the plerocercoid of the cestode Spirometra mansoni is the main causative agent of human and animal sparganosis. However, the population genetic structure of this parasite remains unclear. In this study, we genotyped S. mansoni isolates with the aim to improve current knowledge on the evolution and population diversity of this cestode. We first screened 34 perfect simple sequence repeats (SSRs) using all available omic data and then constructed target sequencing technology (Target SSR-seq) based on the Illumina NovaSeq platform. Next, a series of STRUCTURE. clustering, principal component, analysis of molecular variance and TreeMix analyses were performed on 362 worm samples isolated from 12 different hosts in 16 geographical populations of China to identify the genetic structure. A total of 170 alleles were detected. The whole population could be organized and was found to be derived from the admixture of two ancestral clusters. TreeMix analysis hinted that possible gene flow occurred from Guizhou (GZ) to Sichuan (SC), SC to Jaingxi (JX), SC to Hubei (HB), GZ to Yunnan (YN) and GZ to Jiangsu (JS). Both neighbor-joining clustering and principal coordinate analysis showed that isolates from intermediate hosts tend to cluster together, while parasites from definitive hosts revealed greater genetic differences. Generally, a S. mansoni population was observed to harbor high genetic diversity, moderate genetic differentiation and a little genetic exchange among geographical populations. A Target SSR-seq genotyping method was successfully developed, and an in-depth view of genetic diversity and genetic relationship will have important implications for the prevention and control of sparganosis.
作者机构：
[Li, Rong; Deng, Yuan-Ping; Liu, Guo-Hua; Wang, Hui-Mei] Hunan Agr Univ, Coll Vet Med, Res Ctr Parasites & Vectors, Changsha 410128, Hunan, Peoples R China.;[Tu, Ya] Beijing Wildlife Rescue & Rehabil Ctr, Beijing 101300, Peoples R China.
通讯机构：
[Ya Tu] B;[Yuan-Ping Deng] R;Beijing Wildlife Rescue and Rehabilitation Center, Beijing, 101300, China<&wdkj&>Research Center for Parasites & Vectors, College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan province, 410128, China
通讯机构：
[Jine Yi; Lijuan Zhu] H;Hunan Engineering Research Center of Livestock and Poultry Health Care, College of Veterinary Medicine, Hunan Agricultural University, Changsha, China
通讯机构：
[Zhihang Yuan; Jine Yi] H;Hunan Engineering Research Center of Livestock and Poultry Health Care, College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China
通讯机构：
[Zhao-Ying Liu; Zhao-Ying Liu Zhao-Ying Liu Zhao-Ying Liu] C;College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan, China<&wdkj&>Hunan Engineering Technology Research Center of Veterinary Drugs, Hunan Agricultural University, Changsha, Hunan, China
作者机构：
[Yan, Fen; Zhang, Lu; Wu, Zhi-feng; Duan, De-yong; Cheng, Tian-yin; Liu, Lei] Hunan Agr Univ, Coll Vet Med, Res Ctr Parasites Vectors RCPV, Changsha 410128, Peoples R China.
通讯机构：
[Tian-yin Cheng] R;Research Center for Parasites & Vectors (RCPV), College of Veterinary Medicine, Hunan Agricultural University, Changsha, China
摘要：
Inactivated Mycoplasma hyopneumoniae vaccine is used extensively to control M. hyopneumoniae infection worldwide. Quantification techniques are essential in the process of standardizing and validating vaccines. In this study, we developed and optimized an indirect competitive enzyme linked immunosorbent assay (ic-ELISA) for the rapid quantification of M. hyopneumoniae antigen during vaccine production. Briefly, whole M. hyopneumoniae antigen was coated onto microtiter plates, and a polyclonal antibody against M. hyopneumoniae recombinant elongation factor thermo unstable (EF-Tu) protein was prepared and added with the samples to be tested. The methods were optimized and showed significant reproducibility, with coefficients of variation of 4.01% and 6.14% for the intra-and inter-assays, respectively. Quantification of M. hyopneumoniae cultures at different growth stages using the ic-ELISA test showed a similar curve to that of the traditional color changing units (CCU) assay, with a delay in the time when the amount reached the peak and started to fall. In the inactivated vaccine production process, the cultures could be harvested later than that for the live vaccine, at about 12 h after the end of the logarithmic growth phase. Different batches of cultures were measured for their relative potency value compared with the in-house reference vaccine, which was used to determine whether the cultures met the antigen amount requirements for vaccine preparation. The curves of the CCU titer and ic-ELISA titer in the logarithmic phase correlated strongly and a linear regression equation was established to calculate the CCU values rapidly using the ic-ELISA results. In conclusion, an ic-ELISA method was established to rapidly assess the amount of antigen in an M. hyopneumoniae culture during the vaccine production process.
通讯机构：
[Jine Yi] H;Hunan Engineering Research Center of Livestock and Poultry Health Care, College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, PR China<&wdkj&>Hunan Collaborative Innovation Center of Animal Production Safety, Changsha 410128, PR China
摘要：
Pseudorabies virus (PRV) is considered as an emerging zoonotic pathogen since its isolation from a human case. Meanwhile, the disease caused by PRV infection has led huge economic losses to Chinese pig industry since 2011. In this study, we constructed a recombinant PRV stably expressing the enhanced green fluorescent protein (EGFP) by homologous recombination technology for evaluating its susceptibility to different human cell lines and screening antiviral compounds. Stably expressed EGFP by this designed rPRVHuN-EGFP virus was confirmed in the infected cells, moreover, the growth kinetics of which was similar to that of wild type strain. Importantly, the application of this rPRV allowed us to easily verify its infectivity in all tested human cell lines, although the infection efficiencies were lower than that in PK15 cells. Meanwhile, the antiviral activities of harmine and PHA767491 were also conveniently validated in vitro, as directly reflected by the reduced EGFP signals. These results demonstrate that this recombinant PRV virus should be a useful tool for basic virology researches and antiviral agent screening.
作者机构：
[Li, Qien; Dongzhi, Duojie; Zhong, Jiangbin; Wan, Digao; Guo, Xiao] Qinghai Univ, Tibetan Medicial Coll, Tibetan Med Res Ctr, State Key Lab Tibetan Med Res & Dev, Xining 810016, Qinghai, Peoples R China.;[Huang, Meizhou] Affiliated Hosp Southwest Med Univ, Acad Expert Workstat Sichuan Prov, Luzhou 646000, Peoples R China.;[Dong, Zhen] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Peoples R China.
通讯机构：
[Meizhou Huang] A;Academician (Expert) Workstation of Sichuan Province, The Affiliated Hospital of Southwest Medical University, Luzhou 646000, China
摘要：
BACKGROUND: Rhododendron nivale Hook. f (R.n), one of the four Manna Stash used in Tibetan medicine to delay aging, possesses anti-aging pharmacological activity. However, which R.n ingredients contain anti-aging properties and the underlying mechanisms involved are unclear. HYPOTHESIS/PURPOSE: Based on interactions between gut microbiota and natural medicines and the important role of gut microbiota in anti-aging, the study investigated the hypothesis that R.n possesses anti-aging properties and the interaction of gut microbiota with R.n is responsible for its anti-aging effects. STUDY DESIGN: The primary active ingredients of R.n and their target function and pathway enrichment were explored. An aging mouse model was used to clarify the underlying anti-aging mechanisms of R.n. METHODS: Chromatography, spectroscopy, nuclear magnetic technology, and pharmacology were used to reveal the major active ingredients of ethanol extract residues of R.n (RNEA). The target function and pathway enrichment of these active ingredients were explored. Plasma metabolomics coupled with intestinal flora evaluation and bioinformatics analysis was used to clarify the underlying anti-aging mechanisms of RNEA. RESULTS: Myricetin-3-β-D-xylopyranoside, hyperin, goospetin-8-methyl ether 3-β-D-galactoside, and diplomorphanin B were separated and identified from RNEA. The network pharmacology study revealed that the active ingredients' target function and pathway enrichment focused mainly on the glutathione antioxidant system. In a D-galactose-induced mouse model of aging, RNEA was shown to possess suitable anti-aging pharmacological activity, as indicated by the amelioration of memory loss and weakened superoxide dismutase and glutathione peroxidase activities. Plasma metabolomics coupled with intestinal flora examination and bioinformatics analysis revealed that RNEA could regulate the expression of glutathione-related enzymes and ameliorate D-galactose-induced imbalances in methionine, glycine, and serine, and betaine and galactose metabolism. The results showed that RNEA reshaped the disordered intestinal flora and mitigated the D-galactose-mediated decline in glutathione oxidase expression, further confirming that the anti-aging effect of RNEA was closely related to regulation of the glutathione antioxidant system. CONCLUSION: RNEA, consisting of myricetin-3-β-D-xylopyranoside, hyperin, goospetin-8-methyl ether 3-β-D-galactoside, and diplomorphanin B, possesses anti-aging activity. The anti-aging effect of RNEA might be due to reshaping intestinal flora homeostasis, increasing the expression of glutathione peroxidase 4 in the intestines and liver, enhancing glutathione peroxidase activity, and reinforcing the glutathione antioxidant system.
摘要：
Toxoplasma gondii (T. gondii) is a zoonotic intracellular protozoan parasite that can invade, replicate and survive in almost all cells of warm-blooded animals. T. gondii infection threatens the life of the fetus or can cause morbidity in the infant. As the only definitive host of T. gondii, felids spread the pathogen mainly by forming oocysts in the small intestines and discharging the oocysts into the ambient environment, consequently polluting water, vegetables, and meat products. In this study, we used untargeted metabolomics technology to study the changes in metabolites that occurred during the early stage of oocyst formation in the cat small intestine following T. gondii infection and attempted to identify metabolic biomarkers that could potentially be used as diagnostic molecular markers in the future. Domestic cats (Felis catus) were infected with T. gondii Pru tissue cysts, and samples of their small intestinal epithelium were collected at 2 and 4 days post-infection (DPI) for metabolic analysis. LC-MS/MS and multivariate statistical analysis were employed to detect metabolomic signatures that discriminated between the infected and control groups. A total of 1673 ions and 1201 ions were obtained in the positive and negative modes, respectively. Of these ions, 175 were up-regulated and 127 were down-regulated in the positive ion mode; whereas, 123 were up-regulated and 81 were down-regulated in the negative ion mode. Three commonly altered ions (0.74_313.0414m/z, 8.82_615.2621m/z and 8.16_325.2362m/z) were determined to have potential research value. Seventy common metabolic pathways were enriched at two time points, with arginine biosynthesis, pyrimidine metabolism, pantothenate and CoA biosynthesis being the three most significant pathways related to T. gondii. The area under the curve (AUC) of differential metabolites combined with relevant literature analysis showed that N-Methylpelletierine and 3,3-Difluoro-17-methyl-5alpha-androstan-17beta-ol have higher predictability and better potential application value than other metabolites. Our analysis of metabolic markers during the early stage of T. gondii oocyst formation in the small intestine of the definitive host (cat) provided novel insight for understanding oocyst development and a theoretical basis for the application of potential biomarkers.
作者机构：
[Xiao, Hong-Bo; Wang, Yi-Shan; Tian, Miao-Miao] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Peoples R China.
通讯机构：
[Hong-Bo Xiao] C;College of Veterinary Medicine, Hunan Agricultural University, Changsha, China
关键词：
Stomatitis;Angiopoietin-like protein 4;Nuclear transcription factor-kappa B;Peroxisome proliferator-activated receptor alpha;Stomatitis;Mice
摘要：
BACKGROUND: Stomatitis is inflammation of the oral mucosa. Angiopoietin-like protein 4 (ANGPTL4) has pleiotropic functions both anti-inflammatory and pro-inflammatory properties. In the present study, we tested whether there is a correlation between increased ANGPTL4 expression and inflammation in stomatitis mice and the mechanisms involved. METHODS AND RESULTS: In this study, the oral mucosa of mice was burned with 90% phenol and intraperitoneal injection of 5-fluorouracil to establish the model of stomatitis mice. The pathological changes of stomatitis mice were observed by H&E staining of paraffin section. The expressions of cytokines and ANGPTL4 were detected by fluorescence quantitative PCR, and the protein levels of ANGPTL4 were detected by western blot. Compared with control group, the oral mucosal structure of model mice was damaged. The expression of ANGPTL4 were significantly increased concomitantly with elevated production of anti-inflammatory cytokine (peroxisome proliferator-activated receptoralpha) and pro-inflammatory cytokines [nuclear transcription factor-kappaB, interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α] in mice with stomatitis. CONCLUSIONS: This study suggests that ANGPTL4 may be a double-edged sword in multiple inflammatory responses in stomatitis mice.
期刊：
World Journal of Microbiology and Biotechnology,2022年38(6):1-10 ISSN：0959-3993
通讯作者：
Zeng, JG;Huang, P
作者机构：
[Sun, Mengshan; Zeng, Jianguo; Xu, Zixuan] Hunan Agr Univ, Hunan Key Lab Tradit Chinese Vet Med, Changsha, Hunan, Peoples R China.;[Zhong, Xiaohong; Sun, Mengshan] Hunan Agr Univ, Coll Hort, Changsha, Hunan, Peoples R China.;[Zhou, Li] Hunan Acad Agr Sci, Hunan Inst Agr Environm & Ecol, Changsha, Hunan, Peoples R China.;[Xu, Zixuan] Hunan Univ Chinese Med, Sch Pharm, Changsha, Hunan, Peoples R China.;[Huang, P; Huang, Peng] Hunan Agr Univ, Coll Anim Sci & Technol, Changsha, Hunan, Peoples R China.
通讯机构：
[Zeng, JG ; Huang, P ] H;Hunan Agr Univ, Hunan Key Lab Tradit Chinese Vet Med, Changsha, Hunan, Peoples R China.;Hunan Agr Univ, Coll Anim Sci & Technol, Changsha, Hunan, Peoples R China.;Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.;Hunan Agr Univ, Natl & Local Union Engn Res Ctr Vet Herbal Med Re, Changsha, Hunan, Peoples R China.
作者机构：
[Zhang, Yu; Fu, Yi-Tian; Deng, Yuan-Ping; Nie, Yu] Research Center for Parasites & Vectors, College of Veterinary Medicine, Hunan Agricultural University, Changsha, 410128, Hunan province, China;[Yao, Chaoqun] Department of Biomedical Sciences and One Health Center for Zoonoses and Tropical Veterinary Medicine, Ross University School of Veterinary Medicine, P.O. Box 334, Basseterre, St. Kitts and Nevis;[Liu, Guo-Hua] Research Center for Parasites & Vectors, College of Veterinary Medicine, Hunan Agricultural University, Changsha, 410128, Hunan province, China. liuguohua5202008@163.com
通讯机构：
[Guo-Hua Liu] R;Research Center for Parasites & Vectors, College of Veterinary Medicine, Hunan Agricultural University, Hunan province, Changsha, China
摘要：
Fleas (Insecta: Siphonaptera) are obligatory hematophagous ectoparasites of humans and animals and serve as vectors of many disease-causing agents. Despite past and current research efforts on fleas due to their medical and veterinary importance, correct identification and robust phylogenetic analysis of these ectoparasites have often proved challenging. We decoded the complete mitochondrial (mt) genome of the human flea Pulex irritans and nearly complete mt genome of the dog flea Ctenocephalides canis, and subsequently used this information to reconstruct the phylogeny of fleas among Endopterygota insects. The complete mt genome of P. irritans was 20,337 bp, whereas the clearly sequenced coding region of the C. canis mt genome was 15,609 bp. Both mt genomes were found to contain 37 genes, including 13 protein-coding genes, 22 transfer RNA genes and two ribosomal RNA genes. The coding region of the C. canis mt genome was only 93.5% identical to that of the cat flea C. felis, unequivocally confirming that they are distinct species. Our phylogenomic analyses of the mt genomes showed a sister relationship between the order Siphonaptera and orders Diptera + Mecoptera + Megaloptera + Neuroptera and positively support the hypothesis that the fleas in the order Siphonaptera are monophyletic. Our results demonstrate that the mt genomes of P. irritans and C. canis are different. The phylogenetic tree shows that fleas are monophyletic and strongly support an order-level objective. These mt genomes provide novel molecular markers for studying the taxonomy and phylogeny of fleas in the future.
