1.Screening and identification of host proteins interacting with the non-structural protein 15 (Nsp15) of porcine epidemic diarrhea virus.
Jinlei SUN ; Ruiming YU ; Liping ZHANG ; Zhongwang ZHANG ; Yonglu WANG ; Li PAN ; Quanwei ZHANG ; Xinsheng LIU
Chinese Journal of Biotechnology 2024;40(12):4533-4545
To screen and identify the key host proteins interacting with the non-structural protein 15 (Nsp15) of porcine epidemic diarrhea virus (PEDV). The IP/pull-down assay and mass spectrometry were employed to screen and identify the host proteins interacting with Nsp15. The interaction between the host protein and Nsp15 was studied by co-immunoprecipitation and laser scanning confocal microscopy. Finally, Western blotting and RT-qPCR were employed to examine the interaction between SLC25a3 and PEDV. The recombinant eukaryotic expression vector pcDNA3.1(+)-Flag-Nsp15 was successfully constructed, and the host protein SLC25a3 interacting with PEDV Nsp15 was screened out. An interaction existed between SLC25a3 and Nsp15, and SLC25a3 significantly inhibited PEDV replication in a dose-dependent manner. SLC25a3 inhibits PEDV replication. The results of this study provide a basis for deciphering the role and mechanism of SLC25a3 in the host immune response to PEDV infection.
Porcine epidemic diarrhea virus/genetics*
;
Viral Nonstructural Proteins/metabolism*
;
Animals
;
Swine
;
Virus Replication
;
Coronavirus Infections/veterinary*
;
Swine Diseases/metabolism*
2.Prevalence of Echinococcus infections in wild carnivores based on copro - DNA tests in Serthar County of Sichuan Province.
L YANG ; Y YANG ; W YU ; Q WANG ; B ZHONG ; K HUA ; Y LIU ; Y HUANG
Chinese Journal of Schistosomiasis Control 2023;35(5):492-496
OBJECTIVE:
To investigate the prevalence of Echinococcus infections in wild carnivores in Serthar County, Sichuan Province, so as to provide insights into echinococcosis control in local areas.
METHODS:
Stool samples were collected from wild carnivores in Serthar County, Sichuan Province in May 2021, and the host sources of stool samples and Echinococcus infections were identified using PCR assays. The prevalence of E. multilocularis, E. granulosus and E. shiquicus infections was estimated in different hosts.
RESULTS:
A total of 583 stool samples were collected from wild carnivores, including 147 stool samples from fox, 154 from wolf, 227 from wild dogs and 11 from lynx. The overall prevalence of E. multilocularis, E. granulosus and E. shiquicus infections was 5.68%, 0.19% and 14.20% in canine stool samples, and no E. granulosus infection was detected in fox stool samples, while the prevalence of E. multilocularis and E. shiquicus infections was 0.68% and 47.62% in fox stool samples (χ2 = 88.41, P < 0.001). No E. granulosus or E. shiquicus infection was detected in wolf stool samples, and the prevalence of E. multilocularis infection was 10.39% in wolf stool samples. The prevalence of E. multilocularis, E. granulosus and E. shiquicus infections was 5.73%, 0.44% and 2.20% in canine stool samples (χ2 = 12.13, P < 0.01). In addition, the prevalence of E. multilocularis infections was significantly higher in wolf stool samples than in canine and fox stool samples (χ2 = 13.23, P < 0.01), and the prevalence of E. shiquicus infections was significantly higher in fox stool samples than in canine and wolf stool samples (χ2 = 187.01, P < 0.001). No Echinococcus infection was identified in 11 lynx stool samples.
CONCLUSIONS
The prevalence of Echinococcus infections is high in wild canines in Serthar County, Sichuan Province. Wolf, wild dog and fox all participate in the wild life cycle of E. multilocularis in Serthar County, and wolf and wild dogs may play a more important role.
Animals
;
Dogs/microbiology*
;
China/epidemiology*
;
DNA, Helminth/genetics*
;
Echinococcosis/veterinary*
;
Feces
;
Foxes/microbiology*
;
Lynx/microbiology*
;
Prevalence
;
Wolves/microbiology*
;
Carnivora/microbiology*
3.Molecular detection and subtyping of Blastocystis sp. in pigs in Anhui Province.
S GAO ; J WANG ; X WU ; X LUO ; Q LI ; D CHEN ; X LIU ; W LI
Chinese Journal of Schistosomiasis Control 2023;35(5):508-512
OBJECTIVE:
To investigate the prevalence and subtype distribution of Blastocystis sp. in pigs in Anhui Province.
METHODS:
A total of 500 stool samples were collected from large-scale pig farms in Bozhou, Anqing, Chuzhou, Hefei, Fuyang, and Lu'an cities in Anhui Province from October to December 2015. Blastocystis was detected in pig stool samples using a PCR assay based on the small subunit ribosomal RNA (SSU rRNA) gene, and positive samples were subjected to sequencing and sequence analysis. Blastocystis subtypes were characterized in the online PubMLST database, and verified using phylogenetic tree created with the neighbor-joining algorithm in the Meta software.
RESULTS:
The prevalence of Blastocystis infection was 43.2% (216/500) in pigs in 6 cities of Anhui Province, and all pig farms were tested positive for Blastocystis. There was a region-specific prevalence rate of Blastocystis (17.2% to 50.0%) (χ2 = 26.084, P < 0.01), and there was a significant difference in the prevalence of Blastocystis sp. among nursery pigs (39.6%), preweaned pigs (19.1%), and growing pigs (62.3%) (χ2 = 74.951, P < 0.01). Both online inquiry and phylogenetic analysis revealed ST1, ST3, and ST5 subtypes in pigs, with ST5 as the predominant subtype.
CONCLUSIONS
The prevalence of Blastocystis sp. is high in pigs in Anhui Province, with three zoonotic subtypes identified, including ST1, ST3, and ST5.
Animals
;
Swine
;
Blastocystis/genetics*
;
Phylogeny
;
Blastocystis Infections/veterinary*
;
Polymerase Chain Reaction
;
Prevalence
;
Feces
;
Genetic Variation
4.Rice bran oil supplementation protects swine weanlings against diarrhea and lipopolysaccharide challenge.
Juncheng HUANG ; Wenxia QIN ; Baoyang XU ; Haihui SUN ; Fanghua JING ; Yunzheng XU ; Jianan ZHAO ; Yuwen CHEN ; Libao MA ; Xianghua YAN
Journal of Zhejiang University. Science. B 2023;24(5):430-441
Early weaned piglets suffer from oxidative stress and enteral infection, which usually results in gut microbial dysbiosis, serve diarrhea, and even death. Rice bran oil (RBO), a polyphenol-enriched by-product of rice processing, has been shown to have antioxidant and anti-inflammatory properties both in vivo and in vitro. Here, we ascertained the proper RBO supplementation level, and subsequently determined its effects on lipopolysaccharide (LPS)-induced intestinal dysfunction in weaned piglets. A total of 168 piglets were randomly allocated into four groups of seven replicates (42 piglets each group, (21±1) d of age, body weight (7.60±0.04) kg, and half males and half females) and were given basal diet (Ctrl) or basal diet supplemented with 0.01% (mass fraction) RBO (RBO1), 0.02% RBO (RBO2), or 0.03% RBO (RBO3) for 21 d. Then, seven piglets from the Ctrl and the RBO were treated with LPS (100 μg/kg body weight (BW)) as LPS group and RBO+LPS group, respectively. Meanwhile, seven piglets from the Ctrl were treated with the saline vehicle (Ctrl group). Four hours later, all treated piglets were sacrificed for taking samples of plasma, jejunum tissues, and feces. The results showed that 0.02% was the optimal dose of dietary RBO supplementation based on diarrhea, average daily gain, and average daily feed intake indices in early weaning piglets. Furthermore, RBO protected piglets against LPS-induced jejunal epithelium damage, which was indicated by the increases in villus height, villus height/crypt depth ratio, and Claudin-1 levels, as well as a decreased level of jejunal epithelium apoptosis. RBO also improved the antioxidant ability of LPS-challenged piglets, which was indicated by the elevated concentrations of catalase and superoxide dismutase, and increased total antioxidant capacity, as well as the decreased concentrations of diamine oxidase and malondialdehyde in plasma. Meanwhile, RBO improved the immune function of LPS-challenged weaned piglets, which was indicated by elevated immunoglobulin A (IgA), IgM, β-defensin-1, and lysozyme levels in the plasma. In addition, RBO supplementation improved the LPS challenge-induced dysbiosis of gut microbiota. Particularly, the indices of antioxidant capacity, intestinal damage, and immunity were significantly associated with the RBO-regulated gut microbiota. These findings suggested that 0.02% RBO is a suitable dose to protect against LPS-induced intestinal damage, oxidative stress, and jejunal microbiota dysbiosis in early weaned piglets.
Male
;
Female
;
Swine
;
Animals
;
Lipopolysaccharides/toxicity*
;
Antioxidants/pharmacology*
;
Rice Bran Oil
;
Dysbiosis
;
Dietary Supplements
;
Diarrhea/veterinary*
;
Weaning
;
Body Weight
5.Pteropine Orthoreovirus (PRV7S) does not Establish Persistent Infection in NP460 Cells but with Inflammatory Response Triggered.
Mun Khin Alson LOH ; Zhen Yun SIEW ; Pooi Pooi LEONG ; Rhun Yian KOH ; Soi Moi CHYE ; Siew Tung WONG ; Kenny VOON
Biomedical and Environmental Sciences 2023;36(9):886-891
6.Effects of Aeriscardovia aeriphila on growth performance, antioxidant functions, immune responses, and gut microbiota in broiler chickens.
Muhammad Zahid FAROOQ ; Xinkai WANG ; Xianghua YAN
Journal of Zhejiang University. Science. B 2023;24(11):1014-1026
Aeriscardovia aeriphila, also known as Bifidobacterium aerophilum, was first isolated from the caecal contents of pigs and the faeces of cotton-top tamarin. Bifidobacterium species play important roles in preventing intestinal infections, decreasing cholesterol levels, and stimulating the immune system. In this study, we isolated a strain of bacteria from the duodenal contents of broiler chickens, which was identified as A. aeriphila, and then evaluated the effects of A. aeriphila on growth performance, antioxidant functions, immune functions, and gut microbiota in commercial broiler chickens. Chickens were orally gavaged with A. aeriphila (1×109 CFU/mL) for 21 d. The results showed that A. aeriphila treatment significantly increased the average daily gain and reduced the feed conversion ratio (P<0.001). The levels of serum growth hormone (GH) and insulin-like growth factor 1 (IGF-1) were significantly increased following A. aeriphila treatment (P<0.05). Blood urea nitrogen and aspartate aminotransferase levels were decreased, whereas glucose and creatinine levels increased as a result of A. aeriphila treatment. Furthermore, the levels of serum antioxidant enzymes, including catalase (P<0.01), superoxide dismutase (P<0.001), and glutathione peroxidase (P<0.05), and total antioxidant capacity (P<0.05) were enhanced following A. aeriphila treatment. A. aeriphila treatment significantly increased the levels of serum immunoglobulin A (IgA) (P<0.05), IgG (P<0.01), IgM (P<0.05), interleukin-1 (IL-1) (P<0.05), IL-4 (P<0.05), and IL-10 (P<0.05). The broiler chickens in the A. aeriphila group had higher secretory IgA (SIgA) levels in the duodenum (P<0.01), jejunum (P<0.001), and cecum (P<0.001) than those in the control group. The messenger RNA (mRNA) relative expression levels of IL-10 (P<0.05) and IL-4 (P<0.001) in the intestinal mucosa of chickens were increased, while nuclear factor-κB (NF-κB) (P<0.001) expression was decreased in the A. aeriphila group compared to the control group. Phylum-level analysis revealed Firmicutes as the main phylum, followed by Bacteroidetes, in both groups. The data also found that Phascolarctobacterium and Barnesiella were increased in A. aeriphila-treated group. In conclusion, oral administration of A. aeriphila could improve the growth performance, serum antioxidant capacity, immune modulation, and gut health of broilers. Our findings may provide important information for the application of A. aeriphila in poultry production.
Animals
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Swine
;
Antioxidants/pharmacology*
;
Chickens
;
Gastrointestinal Microbiome
;
Interleukin-10/pharmacology*
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Interleukin-4/pharmacology*
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NF-kappa B/metabolism*
;
Immunity
;
Diet/veterinary*
;
Animal Feed/analysis*
;
Dietary Supplements/analysis*
7.Preparation and immunogenicity evaluation of mRNA vaccine against porcine epidemic diarrhea.
Limin YANG ; Junhong WANG ; Mingguo XU ; Hu WANG ; Xiaojuan ZHANG ; Wenjun LIU ; Chuangfu CHEN
Chinese Journal of Biotechnology 2023;39(7):2624-2633
Porcine epidemic diarrhea (PED) is a highly contagious disease that causes high mortality in suckling piglets. Although several licensed inactivated and live attenuated vaccines were widely used, the infection rate remains high due to unsatisfactory protective efficacy. In this study, mRNA vaccine candidates against PED were prepared, and their immunogenicity was evaluated in mice and pregnant sows. The mRNA PED vaccine based on heterodimer of viral receptor binding region (RBD) showed good immunogenicity. It elicited robust humoral and cellular immune responses in mice, and the neutralizing antibody titer reached 1:300 after a single vaccination. Furthermore, it induced neutralizing antibody level similar to that of the inactivated vaccine in pregnant sows. This study developed a new design of PED vaccine based on the mRNA-RBD strategy and demonstrated the potential for clinical application.
Pregnancy
;
Swine
;
Animals
;
Female
;
Mice
;
Antibodies, Viral
;
Swine Diseases/epidemiology*
;
Viral Vaccines/genetics*
;
Antibodies, Neutralizing
;
Vaccines, Attenuated
;
Diarrhea/veterinary*
8.Exploration of the UMOOC-based blended teaching in veterinary immunology.
Luomeng CHAO ; Guiying KANG ; Kai LIU
Chinese Journal of Biotechnology 2022;38(5):2033-2041
The study was based on the teaching effect of blended teaching of the course of Veterinary Immunology on the U-Massive Open Online Course (UMOOC) platform of Inner Mongolia Minzu University. The students of two classes majoring in veterinary medicine in Inner Mongolia Minzu University were selected to form the traditional teaching group (control group) and the blended teaching group (experimental group), respectively, and the teaching effects were further compared and analyzed. The blended teaching module improved students' active learning attitude along with their ability to grasp knowledge in class, and most students supported to use blended teaching in the future. In conclusion, the UMOOC-based blended teaching methodology stimulated students' active learning attitude toward veterinary immunology and helped to foster innovative consciousness and self-management ability.
China
;
Curriculum
;
Education, Veterinary
;
Humans
;
Learning
;
Students
;
Universities
9.Genetic characteristics of hantavirus detected in rodents in Shenzhen.
Yao LUO ; Yue LI ; Ya Lan HUANG ; Xiao Min ZHANG ; Ling Hong XIONG ; Ren Li ZHANG ; Fan YANG
Chinese Journal of Epidemiology 2022;43(11):1804-1810
Objective: To study the molecular epidemiological characteristics and genotypes of hantavirus carried by rodents in Shenzhen. Methods: Rodents were captured, and their lung samples were collected and grinded for RNA extraction. The hantavirus positive samples were classified by real-time fluorescence PCR. Rat lung nucleic acid samples were selected to amplify the nucleotide sequences of partial M fragments (G2 segment) and S fragments by reverse transcription-nested polymerase chain reaction (RT-nested PCR). The PCR products were then sequenced and homology and phylogenetic tree analyses were conducted. Results: A total of 200 rodents were captured, including 189 Rattus norvegicus, 9 Rattus flavipectus and 2 Mus musculus. The positive rate of hantavirus was 21.0% (42/200), all of the isolates were seoul virus (SEOV) strains. The positive rate of hantavirus in Bao'an district was highest (45.7%), and the difference in detection rate among districts were significant (χ2=25.60,P<0.05). A total of 25 G2 segment sequences and S fragment sequences of SEOV were obtained by virus gene sequencing, and their nucleotide homology was 95.3%-100.0% and 97.6%-100.0%, respectively. Compared with other reference sequences of S2 subtype, the nucleotide homology between the sample sequence and the reference sequence from Guangzhou was high. Analysis on nucleotide homology and phylogenetic tree showed that hantavirus carried by the rodents captured in Shenzhen belonged to SEOV S2 subtype. Analysis on amino acid variation sites revealed that there was a variation in the nucleocapsid protein encoded by S gene from Alanine to Threonine at the 973 position of BA-111. Conclusion: Hantavirus carried by rodents in Shenzhen belongs to S2 subtype of Seoul virus, which have little variation compared with the hantavirus strains obtained in other years in Shenzhen and surrounding provinces.
Mice
;
Rats
;
Animals
;
Orthohantavirus/genetics*
;
Rodentia
;
Phylogeny
;
Hantavirus Infections/veterinary*
;
Communicable Diseases
;
Nucleotides
;
Real-Time Polymerase Chain Reaction
10.Immunization against porcine epidemic diarrhea virus and vaccine development.
Shijuan DONG ; Chunfang XIE ; Fusheng SI ; Bingqing CHEN ; Ruisong YU ; Zhen LI
Chinese Journal of Biotechnology 2021;37(8):2603-2613
Porcine epidemic diarrhea (PED) is a major disease of pigs that inflicts heavy losses on the global pig industry. The etiologic agent is the porcine epidemic diarrhea virus (PEDV), which is assigned to the genus Alphacoronavirus in the family Coronaviridae. This review consists of five parts, the first of which provides a brief introduction to PEDV and its epidemiology. Part two outlines the passive immunity in new born piglets and the important role of colostrum, while the third part summarizes the characteristics of the immune systems of pregnant sows, discusses the concept of the "gut-mammary gland-secretory IgA(sIgA) axis" and the possible underpinning mechanisms, and proposes issues to be addressed when designing a PEDV live vaccine. The final two parts summarizes the advances in the R&D of PEDV vaccines and prospects future perspectives on prevention and control of PEDV, respectively.
Animals
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Antibodies, Viral
;
Coronavirus Infections/veterinary*
;
Female
;
Immunization
;
Porcine epidemic diarrhea virus
;
Pregnancy
;
Swine
;
Swine Diseases/prevention & control*
;
Viral Vaccines

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