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1.
Sci Rep ; 14(1): 13010, 2024 06 06.
Article in English | MEDLINE | ID: mdl-38844489

ABSTRACT

Studies on Bdellovibrio and like organisms (BALOs), obligate predatory bacteria, have highlighted the possibility of regulating bacteria and biofilms; however, yak-derived BALOs are yet to be reported. We aimed to characterize the BALOs isolated and identified from yak (Bos grunniens) feces and examine application potential. BALOs were isolated from healthy yak fecal samples, with Escherichia coli (ATCC 25922) as prey using the double-layer agar method, identified by transmission electron microscopy (TEM), and the specific 16S rDNA sequencing analysis. Sequencing of the 16S rDNA gene indicated that this isolate was 91% similar to the Bdellovibrio sp. NC01 reference strain and was named YBD-1. Proportion of YBD-1 lysed bacteria is 12/13. The YBD-1 showed best growth at 25-40°C, 0-0.25% (w/v) NaCl, and pH 6.5-7.5. YBD-1 significantly reduced the planktonic cells and biofilms of E.coli in co-culture compared to the E.coli group. Additionally, SEM analysis indicated that YBD-1 significantly reduced biofilm formation in E. coli. Furthermore, quantitative Real Time-polymerase chain reaction (qRT-PCR) showed that the expression of the virulence genes fim and iroN and the genes pgaABC involved in biofilm formation went down significantly. We concluded that YBD-1 may have the potential to control bacterial growth and biofilm-associated bacterial illnesses.


Subject(s)
Bdellovibrio , Biofilms , Escherichia coli , Feces , Animals , Feces/microbiology , Cattle , Biofilms/growth & development , Escherichia coli/genetics , Escherichia coli/isolation & purification , Bdellovibrio/genetics , Bdellovibrio/isolation & purification , Bdellovibrio/physiology , RNA, Ribosomal, 16S/genetics , Phylogeny
2.
Front Microbiol ; 15: 1388420, 2024.
Article in English | MEDLINE | ID: mdl-38756726

ABSTRACT

Feline calicivirus (FCV) is a prevalent and impactful viral pathogen affecting domestic cats. As an RNA virus, FCV exhibits high mutability and genetic plasticity, enabling its persistence within cat populations. Viral genetic diversity is associated with a broad spectrum of clinical manifestations, ranging from asymptomatic infections and mild oral and upper respiratory tract diseases to the potential development of virulent systemic, and even fatal conditions. This diversity poses distinctive challenges in diagnosis, treatment, and prevention of diseases caused by FCV. Over the past four decades, research has significantly deepened understanding of this pathogen, with an emphasis on molecular biology, evolutionary dynamics, vaccine development, and disease management strategies. This review discusses various facets of FCV, including its genomic structure, evolution, innate immunity, pathogenesis, epidemiology, and approaches to disease management. FCV remains a complex and evolving concern in feline health, requiring continuous research to enhance understanding of its genetic diversity, to improve vaccine efficacy, and to explore novel treatment options.

3.
Front Vet Sci ; 11: 1372032, 2024.
Article in English | MEDLINE | ID: mdl-38681852

ABSTRACT

Porcine reproductive and respiratory syndrome virus (PRRSV) remains one of the major threats to swine industry, resulting in huge economic losses worldwide. Currently, PRRSV has diversified into multiple lineages with characteristics of extensive recombination in China. In this research, three virus strains were isolated and four virus whole genome sequences were generated and analyzed from clinical samples collected in Gansu province of China in 2023. The four virus strains were designated GSTS4-2023, GSLX2-2023, GSFEI2-2023 and GSBY4-2023. Phylogenetic analysis based on ORF5 sequences showed that GSTS4-2023, GSLX2-2023, GSFEI2-2023 and GSBY4-2023 shared 91.7, 91.2, 93.2 and 92.9% homology with NADC30 strain respectively, and belonged to lineage 1 of PRRSV-2. In addition, one amino acid deletion was observed at position 33 in ORF5 of GSTS4-2023, GSLX2-2023 and GSFEI2-2023. Moreover, amino acid alignment of the four strains showed a typical discontinuous 131-amino acid (aa) deletion in NSP2 for NADC30-like virus strains. Recombination analysis revealed that all four strains originated from NADC30 (lineage 1), with their minor parents coming from JXA1-like strains (lineage 8), VR-2332-like strains (lineage5) and QYYZ-like strains (lineage3). Finally, the three isolated virus strains, GSTS4-2023, GSLX2-2023 and GSFEI2-2023 showed relatively low levels of replication in cell culture. Our findings provide important implications for the field epidemiology of PRRSV.

4.
Vet Sci ; 11(4)2024 Apr 08.
Article in English | MEDLINE | ID: mdl-38668435

ABSTRACT

To investigate the association between 146S antigen contents in FMD inactivated vaccines and levels of antiviral immunity, this study vaccinated 30 kg pigs with three batches of FMD types O and A bivalent inactivated vaccines. Antibody titers and interferon-gamma (IFN-γ) secretion levels were measured on days 7, 14, 21, and 28 after primary immunization and on days 14 and 28 following booster immunization to assess associations between 146S contents and both antibody titers and IFN-γ secretion levels. Furthermore, 30 kg pigs were vaccinated with 46 batches of FMD type O inactivated vaccines and challenged on day 28, after which PD50 values were determined to evaluate the association between 146S content and PD50. The findings suggested that antibody titers and IFN-γ secretion levels at specific time points after immunization were positively associated with 146S contents. Additionally, 146S content showed a positive correlation with PD50, with greater PD50 values recorded for 146S contents ranging from 4.72 to 16.55 µg/dose. This investigation established a significant association between the 146S content in FMD inactivated vaccines and induced immune response against FMDV, thereby emphasizing its critical role in vaccine quality control. The determination of 146S content could serve as a new method for potency testing, offering an alternative to animal challenge tests.

5.
Virology ; 595: 110056, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38552409

ABSTRACT

The Peste des petits ruminant virus (PPRV) is a member of the Paramyxoviridae family and is classified into the genus Measles virus. PPRV predominantly infects small ruminants, leading to mortality rates of nearly 100%, which have caused significant economic losses in developing countries. Host proteins are important in virus replication, but the PPRV nucleocapsid (N) protein-host interacting partners for regulating PPRV replication remain unclear. The present study confirmed the interaction between PPRV-N and the host protein vimentin by co-immunoprecipitation and co-localization experiments. Overexpression of vimentin suppressed PPRV replication, whereas vimentin knockdown had the opposite effect. Mechanistically, N was subjected to degradation via the ubiquitin/proteasome pathway, where vimentin recruits the E3 ubiquitin ligase NEDD4L to fulfill N-ubiquitination, resulting in the degradation of the N protein. These findings suggest that the host protein vimentin and E3 ubiquitin ligase NEDD4L have an anti-PPRV effect.


Subject(s)
Nucleocapsid Proteins , Peste-des-petits-ruminants virus , Vimentin , Virus Replication , Nucleocapsid Proteins/metabolism , Nucleocapsid Proteins/genetics , Vimentin/metabolism , Vimentin/genetics , Animals , Peste-des-petits-ruminants virus/physiology , Peste-des-petits-ruminants virus/genetics , Peste-des-petits-ruminants virus/metabolism , Humans , Ubiquitination , Host-Pathogen Interactions , HEK293 Cells , Nedd4 Ubiquitin Protein Ligases/metabolism , Nedd4 Ubiquitin Protein Ligases/genetics , Cell Line , Peste-des-Petits-Ruminants/virology , Peste-des-Petits-Ruminants/metabolism , Protein Binding
6.
Int J Mol Sci ; 25(4)2024 Feb 12.
Article in English | MEDLINE | ID: mdl-38396878

ABSTRACT

Porcine epidemic diarrhoea virus (PEDV) is a coronavirus that can cause severe watery diarrhoea in piglets, with high morbidity and mortality rates, seriously hindering the healthy development of the global swine industry. In this study, we isolated a strain of PEDV from Tibetan pigs and named it CH/GS/2022. Subsequently, we screened the apoptosis signals of PEDV-infected IPEC-J2 cells and studied the correlation between apoptosis signals and cell apoptosis. The results showed that different infections of PEDV induced different degrees of apoptosis in cells, and PEDV-induced cell apoptosis was dose-dependent. We then detected the expression of the p53, p38, JNK, Bax, and Bcl-2 genes in the apoptosis signal pathway. The results showed that 24 h after PEDV infection, the expression of the p53, p38, JNK, and Bax genes in IPEC-J2 cells increased significantly, while the expression of the Bcl-2 gene decreased significantly (p < 0.05). Subsequently, we used Western blot to detect the protein levels of these five genes, and the results showed that PEDV infection upregulated the expression of p53, p38, JNK, and Bax proteins (p < 0.05) while downregulating the expression of Bcl-2 protein (p < 0.05). Thus, it was initially inferred that PEDV infection could regulate cell apoptosis by activating the p53, p38, and JNK signalling pathways. Finally, we further investigated the apoptosis of the cells through the use of inhibitors. The results indicated that the p53 inhibitor Pifithrin-α has a significant inhibitory effect on the expression of the p53 protein after PEDV infection and can reverse the expression levels of Bax and Bcl-2 proteins. This suggested that p53 is involved in PEDV-induced cell apoptosis. Similarly, the p38 MAPK inhibitor SB203580 has an inhibitory effect on the expression of the p38 protein and can reverse the expression levels of Bax and Bcl-2 proteins. This suggested that p38 is also involved in PEDV-induced cell apoptosis. On the other hand, the JNK inhibitor SP600125 has no inhibitory effect on the expression of the JNK protein after PEDV infection, but the expression levels of Bax and Bcl-2 proteins have changed. Furthermore, it is noteworthy that SP600125 can inhibit the activity of apoptotic proteins but not their levels, resulting in reduced cell apoptosis. These preliminary results indicated that JNK may be involved in PEDV-induced IPEC-J2 cell apoptosis.


Subject(s)
Anthracenes , Porcine epidemic diarrhea virus , Animals , Swine , Cell Line , Porcine epidemic diarrhea virus/physiology , bcl-2-Associated X Protein/genetics , Tumor Suppressor Protein p53/genetics , Tibet
7.
BMC Vet Res ; 20(1): 4, 2024 Jan 03.
Article in English | MEDLINE | ID: mdl-38172906

ABSTRACT

INTRODUCTION: This study aimed to investigate the microbial characteristics of yak uteri collected using intrauterine cotton swabs (CS) during different reproductive stages and the correlation of these microbial characteristics with reproductive status. METHODS: We used a macrogenomic approach to analyze the functional aspects of different microorganisms in samples collected during the pre-estrus, estrus, late estrus, and diestrus stages. RESULTS: The results revealed the presence of 1293 microbial genera and 3401 microbial species in the uteri of yaks at different reproductive stages. The dominant bacterial species varied across the different periods, with Micrococcus and Proteus being dominant during pre-estrus; Pseudomonas, Clostridium, Flavobacterium, Bacillus, and Staphylococcus during estrus; Acinetobacter, Bacillus and Proteus during late estrus; and Pseudomonas, Escherichia coli, and Proteus during diestrus. DISCUSSION: The primary functions of these bacteria are enriched in various metabolic pathways, including carbohydrate and amino acid metabolism, intracellular transport and secretion, post-translational protein modification, and drug resistance. These findings suggest that the microbial diversity in the uterus of yaks plays a crucial role in reproductive regulation and can help prevent reproductive tract-related diseases.


Subject(s)
Estrus , Uterus , Female , Cattle , Animals , Uterus/metabolism , Reproduction
8.
Microb Pathog ; 181: 106185, 2023 Aug.
Article in English | MEDLINE | ID: mdl-37269880

ABSTRACT

Porcine Epidemic Diarrhoea (PED) is an acute, extremely infectious intestinal disease of pigs caused by the Porcine Epidemic Diarrhoea Virus (PEDV). The virus can affect pigs of all breeds and age groups and shows varying degrees of symptoms, with piglets, in particular, being infected with mortality rates of up to 100%. PEDV was first identified in China in the 1980s and in October 2010 a large-scale PED outbreak caused by a variant of PEDV occurred in China, resulting in huge economic losses. Initially, vaccination can effectively prevent the classical strain, but since December 2010, the PEDV variant has caused "persistent diarrhoea" with severe vomiting, watery diarrhoea, and high morbidity and mortality in newborn piglets as the dominant clinical features, with a significant increase in morbidity and mortality. This indicates that PEDV strains have mutated during evolution and that traditional vaccines no longer provide effective cross-immune protection, so it is necessary to optimize immunization programs and find effective treatments through epidemiological surveys of PEDV to reduce the economic losses caused by infections with mutated strains. This article reviews the progress of research on the aetiology, epidemiological characteristics, genotyping, pathogenesis, transmission routes, and comprehensive control of PEDV infection in China.


Subject(s)
Coronavirus Infections , Dysentery , Porcine epidemic diarrhea virus , Swine Diseases , Animals , Swine , Porcine epidemic diarrhea virus/genetics , Genotype , Coronavirus Infections/epidemiology , Coronavirus Infections/prevention & control , Coronavirus Infections/veterinary , Diarrhea , China/epidemiology , Swine Diseases/epidemiology , Swine Diseases/prevention & control
9.
BMC Genomics ; 24(1): 281, 2023 May 25.
Article in English | MEDLINE | ID: mdl-37231370

ABSTRACT

BACKGROUND: Cyclic nucleotide-gated ion channels (CNGCs) are nonselective cation channels that are ubiquitous in eukaryotic organisms. As Ca2+ channels, some CNGCs have also proven to be K+-permeable and involved in plant development and responses to environmental stimuli. Sugarcane is an important sugar and energy crop worldwide. However, reports on CNGC genes in sugarcane are limited. RESULTS: In this study, 16 CNGC genes and their alleles were identified from Saccharum spontaneum and classified into 5 groups based on phylogenetic analysis. Investigation of gene duplication and syntenic relationships between S. spontaneum and both rice and Arabidopsis demonstrated that the CNGC gene family in S. spontaneum expanded primarily by segmental duplication events. Many SsCNGCs showed variable expression during growth and development as well as in tissues, suggesting functional divergence. Light-responsive cis-acting elements were discovered in the promoters of all the identified SsCNGCs, and the expression of most of the SsCNGCs showed a diurnal rhythm. In sugarcane, the expression of some SsCNGCs was regulated by low-K+ treatment. Notably, SsCNGC13 may be involved in both sugarcane development and its response to environmental stimuli, including response to low-K+ stress. CONCLUSION: This study identified the CNGC genes in S. spontaneum and provided insights into the transcriptional regulation of these SsCNGCs during development, circadian rhythm and under low-K+ stress. These findings lay a theoretical foundation for future investigations of the CNGC gene family in sugarcane.


Subject(s)
Cyclic Nucleotide-Gated Cation Channels , Saccharum , Cyclic Nucleotide-Gated Cation Channels/genetics , Cyclic Nucleotide-Gated Cation Channels/metabolism , Saccharum/genetics , Saccharum/metabolism , Plant Proteins/metabolism , Phylogeny , Nucleotides, Cyclic/metabolism , Gene Expression Regulation, Plant
10.
Microbiol Spectr ; 11(3): e0459922, 2023 06 15.
Article in English | MEDLINE | ID: mdl-37036366

ABSTRACT

Senecavirus A (SVA) is an emerging viral pathogen related to vesicular disease and neonatal mortality in swine, which results in enormous economic losses to the global swine industry. The clinical signs of SVA are indistinguishable from those of other vesicular diseases, such as foot-and-mouth disease, which is an economically devastating animal disease. Therefore, development of a rapid, sensitive, and specific diagnostic method for the detection of SVA infection is critical for the prevention and control of SVA and would help to rule out other exotic diseases. In this study, two whole-porcine anti-SVA antibodies (1M5 and 1M25) were produced using single B cell antibody technology. 1M5 and 1M25 possessed neutralizing activity against SVA but recognized different conformational epitopes that depended on the intact virion. Using 1M5 as the capture antibody and biotinylated 1M25 as the detection antibody, a reliable and rapid competitive enzyme-linked immunosorbent assay for detecting neutralizing antibodies (NAC-ELISA) against SVA was developed. Receiver-operating characteristic curve analysis showed that the sensitivity and specificity of the assay were 98.11% and 100%, respectively, with a cutoff percent inhibition value of 45%. The NAC-ELISA was specific for detecting SVA-specific antibodies, without cross-reactivity to other virus-infected sera. The results of the NAC-ELISA showed a strong agreement with the results of the virus neutralization test. Therefore, the NAC-ELISA developed in this study represents a sensitive, specific, and reliable tool for the detection of SVA-specific antibodies, which is applicable for serodiagnosis and serological surveillance of SVA and is conducive to the prevention and control of SVA. IMPORTANCE Senecavirus A (SVA) is an emerging picornavirus related to vesicular disease and neonatal mortality in swine, which results in enormous economic losses worldwide. Additionally, the clinical characteristics of the disease are indistinguishable from those of other vesicular diseases, such as foot-and-mouth disease. Therefore, developing tools for rapidly and accurately detecting SVA infection is critical and urgent. In this study, two porcine-derived monoclonal antibodies against SVA were generated, and a competitive ELISA for the detection of neutralizing antibodies (NAC-ELISA) against SVA was successfully developed using these two porcine monoclonal antibodies. The NAC-ELISA was SVA specific with no cross-reactivity to other related pathogens and had high sensitivity, specificity, and reproducibility for detecting SVA-specific antibody. Therefore, the NAC-ELISA developed in this study may be of great value as a simple and reliable tool for serodiagnosis or surveillance of SVA and may facilitate the prevention and control of SVA.


Subject(s)
Foot-and-Mouth Disease , Swine Diseases , Animals , Swine , Antibodies, Neutralizing , Antibodies, Monoclonal , Reproducibility of Results , Antibodies, Viral , Enzyme-Linked Immunosorbent Assay/methods , Swine Diseases/epidemiology
11.
BMC Genomics ; 24(1): 79, 2023 Feb 17.
Article in English | MEDLINE | ID: mdl-36800937

ABSTRACT

BACKGROUND: B-box (BBX) zinc-finger transcription factors play crucial roles in plant growth, development, and abiotic stress responses. Nevertheless, little information is available on sugarcane (Saccharum spp.) BBX genes and their expression profiles. RESULTS: In the present study, we characterized 25 SsBBX genes in the Saccharum spontaneum genome database. The phylogenetic relationships, gene structures, and expression patterns of these genes during plant growth and under low-nitrogen conditions were systematically analyzed. The SsBBXs were divided into five groups based on phylogenetic analysis. The evolutionary analysis further revealed that whole-genome duplications or segmental duplications were the main driving force for the expansion of the SsBBX gene family. The expression data suggested that many BBX genes (e.g., SsBBX1 and SsBBX13) may be helpful in both plant growth and low-nitrogen stress tolerance. CONCLUSIONS: The results of this study offer new evolutionary insight into the BBX family members in how sugarcane grows and responds to stress, which will facilitate their utilization in cultivated sugarcane breeding.


Subject(s)
Saccharum , Saccharum/genetics , Saccharum/metabolism , Phylogeny , Plant Breeding , Plant Development , Stress, Physiological/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Gene Expression Regulation, Plant
12.
Virol Sin ; 38(1): 96-107, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36435451

ABSTRACT

African swine fever virus (ASFV) is an important pathogen that causes a highly contagious and lethal disease in swine, for which neither a vaccine nor treatment is available. The DNA repair enzyme 8-oxoguanine DNA glycosylase 1 (OGG1), which excises the oxidative base lesion 8-oxo-7,8-dihydroguanine (8-oxoG), has been linked to the pathogenesis of different diseases associated with viral infections. However, the role of OGG1-base excision repair (BER) in ASFV infection has been poorly investigated. Our study aimed to characterize the alteration of host reactive oxygen species (ROS) and OGG1 and to analyse the role of OGG1 in ASFV infection. We found that ASFV infection induced high levels and dynamic changes in ROS and 8-oxoG and consistently increased the expression of OGG1. Viral yield, transcription level, and protein synthesis were reduced in ASFV-infected primary alveolar macrophages (PAMs) treated by TH5487 or SU0268 inhibiting OGG1. The expression of BER pathway associated proteins of ASFV was also suppressed in OGG1-inhibited PAMs. Furthermore, OGG1 was found to negatively regulate interferon ß (IFN-ß) production during ASFV infection and IFN-ß could be activated by OGG1 inhibition with TH5487 and SU0268, which blocked OGG1 binding to 8-oxoG. Additionally, the interaction of OGG1 with viral MGF360-14-L protein could disturb IFN-ß production to further affect ASFV replication. These results suggest that OGG1 plays the crucial role in successful viral infection and OGG1 inhibitors SU0268 or TH5487 could be used as antiviral agents for ASFV infection.


Subject(s)
African Swine Fever Virus , African Swine Fever , Swine , Animals , African Swine Fever Virus/genetics , Reactive Oxygen Species/metabolism , DNA Repair , Oxidative Stress , Virus Replication
13.
BMC Plant Biol ; 22(1): 510, 2022 Nov 02.
Article in English | MEDLINE | ID: mdl-36319957

ABSTRACT

BACKGROUND: Growth regulating factors (GRFs) are transcription factors that regulate diverse biological and physiological processes in plants, including growth, development, and abiotic stress. Although GRF family genes have been studied in a variety of plant species, knowledge about the identification and expression patterns of GRFs in sugarcane (Saccharum spp.) is still lacking. RESULTS: In the present study, a comprehensive analysis was conducted in the genome of wild sugarcane (Saccharum spontaneum) and 10 SsGRF genes were identified and characterized. The phylogenetic relationship, gene structure, and expression profiling of these genes were analyzed entirely under both regular growth and low-nitrogen stress conditions. Phylogenetic analysis suggested that the 10 SsGRF members were categorized into six clusters. Gene structure analysis indicated that the SsGRF members in the same group were greatly conserved. Expression profiling demonstrated that most SsGRF genes were extremely expressed in immature tissues, implying their critical roles in sugarcane growth and development. Expression analysis based on transcriptome data and real-time quantitative PCR verification revealed that GRF1 and GRF3 were distinctly differentially expressed in response to low-nitrogen stress, which meant that they were additional participated in sugarcane stress tolerance. CONCLUSION: Our study provides a scientific basis for the potential functional prediction of SsGRF and will be further scrutinized by examining their regulatory network in sugarcane development and abiotic stress response, and ultimately facilitating their application in cultivated sugarcane breeding.


Subject(s)
Saccharum , Saccharum/genetics , Gene Expression Regulation, Plant , Phylogeny , Plant Proteins/genetics , Plant Breeding , Nitrogen/metabolism
14.
Viruses ; 14(11)2022 10 31.
Article in English | MEDLINE | ID: mdl-36366509

ABSTRACT

Influenza A virus (IAV) is a globally distributed zoonotic pathogen and causes a highly infectious respiratory disease with high morbidity and mortality in humans and animals. IAV has evolved various strategies to counteract the innate immune response, using different viral proteins. However, the mechanisms are not fully elucidated. In this study, we demonstrated that the nonstructural protein 2 (NS2) of H1N1 IAV negatively regulate the induction of type-I interferon. Co-immunoprecipitation experiments revealed that NS2 specifically interacts with interferon regulatory factor 7 (IRF7). NS2 blocks the nuclear translocation of IRF7 by inhibiting the formation of IRF7 dimers, thereby prevents the activation of IRF7 and inhibits the production of interferon-beta. Taken together, these findings revealed a novel mechanism by which the NS2 of H1N1 IAV inhibits IRF7-mediated type-I interferon production.


Subject(s)
Influenza A Virus, H1N1 Subtype , Influenza A virus , Interferon Type I , Humans , Animals , Influenza A virus/physiology , Interferon Regulatory Factor-7/genetics , Interferon Regulatory Factor-7/metabolism , Influenza A Virus, H1N1 Subtype/metabolism , Host-Pathogen Interactions , Immunity, Innate , Interferon Type I/metabolism , Viral Nonstructural Proteins/metabolism
15.
Front Cell Infect Microbiol ; 12: 984537, 2022.
Article in English | MEDLINE | ID: mdl-36189367

ABSTRACT

In order to evaluate the potential and safety of lactic acid bacteria (LAB) isolated from faeces samples of Ganan yak as probiotic for prevention and/or treatment of yak diarrhea, four strains of LAB including Latilactobacillus curvatus (FY1), Weissella cibaria (FY2), Limosilactobacillus mucosae (FY3), and Lactiplantibacillus pentosus (FY4) were isolated and identified in this study. Cell surface characteristics (hydrophobicity and cell aggregation), acid resistance and bile tolerance, compatibility, antibacterial activity and in vitro cell adhesion tests were also carried out to evaluate the probiotic potential of LAB. The results showed that the four isolates had certain acid tolerance, bile salt tolerance, hydrophobicity and cell aggregation, all of which contribute to the survival and colonization of LAB in the gastrointestinal tract. There is no compatibility between the four strains, so they can be combined into a mixed probiotic formula. Antimicrobial tests showed that the four strains were antagonistic to Escherichia coli, Staphylococcus aureus, and Salmonella typhimurium. Moreover, the in vitro safety of the four isolates were determined through hemolytic analysis, gelatinase activity, and antibacterial susceptibility experiments. The results suggest that all the four strains were considered as safe because they had no hemolytic activity, no gelatinase activity and were sensitive to most antibacterial agents. Moreover, the acute oral toxicity test of LAB had no adverse effect on body weight gain, food utilization and organ indices in Kunming mice. In conclusion, the four LAB isolated from yak feces have considerable potential to prevent and/or treat yak bacterial disease-related diarrhea.


Subject(s)
Bacterial Infections , Lactobacillales , Probiotics , Animals , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/pharmacology , Cattle , Diarrhea/veterinary , Feces/microbiology , Lactobacillales/metabolism , Mice
16.
EBioMedicine ; 82: 104177, 2022 Aug.
Article in English | MEDLINE | ID: mdl-35843171

ABSTRACT

BACKGROUND: Alveolar echinococcosis (AE), which is caused by larval Echinococcus multilocularis, is one of the world's most dangerous neglected diseases. Currently, no fully effective treatments are available to cure this disease. METHODS: In vitro protoscolicidal assay along with in vivo murine models was applied in repurposing drugs against AE. Genome-wide identification and homology-based modeling were used for predicting drug targets. RNAi, enzyme assay, and RNA-Seq analyses were utilized for investigating the roles in parasite survival and validations for the drug target. FINDINGS: We identified nelfinavir as the most effective HIV protease inhibitor against larval E. multilocularis. Once-daily oral administration of nelfinavir for 28 days resulted in a remarkable reduction in parasite infection in either immune-competent or immunocompromised mice. E. multilocularis DNA damage-inducible 1 protein (EmuDdi1) is predicted as a target candidate for nelfinavir. We proved that EmuDdi1 is essential for parasite survival and protein excretion and acts as a functionally active protease for this helminth. We found nelfinavir is able to inhibit the proteolytic activity of recombinant EmuDdi1 and block the EmuDdi1-related pathways for protein export. With other evidence of drug efficacy comparison, our results suggest that inhibition of EmuDdi1 is a mechanism by which this HIV proteinase inhibitor mediates its antiparasitic action on echinococcosis. INTERPRETATION: This study demonstrates that nelfinavir is a promising candidate for treating echinococcosis. This drug repurposing study proves that the widely prescribed drug for AIDS treatment is potent in combating E. multilocularis infection and thus provides valuable insights into the development of single-drug therapy for highly prevalent co-infection between HIV and helminth diseases. FUNDING: This work was supported by the National Natural Science Foundation of China (31802179), the Natural Science Foundation of Gansu Province, China (No. 21JR7RA027), and the State Key Laboratory of Veterinary Etiological Biology (No. SKLVEB2021YQRC01).


Subject(s)
Echinococcosis , Echinococcus multilocularis , HIV Protease Inhibitors , Animals , Echinococcosis/drug therapy , Echinococcus multilocularis/genetics , Enzyme Inhibitors/pharmacology , HIV Protease Inhibitors/pharmacology , Mice , Nelfinavir/pharmacology , Pharmaceutical Preparations
17.
Sci Rep ; 12(1): 6597, 2022 04 21.
Article in English | MEDLINE | ID: mdl-35449433

ABSTRACT

Tibetan sheep have lived on the Qinghai-Tibetan Plateau for thousands of years and have good adaptability to the hypoxic environment and strong disease resistance. However, the molecular mechanism by which Tibetan sheep adapt to this extreme environment, especially the role of genetic regulation, is still unknown. Emerging evidence suggests that long noncoding RNAs (lncRNAs) participate in the regulation of a diverse range of biological processes. To explore the potential lncRNAs involved in the adaptation to high-altitude hypoxia of Tibetan sheep, we analysed the expression profile of lncRNAs and mRNAs in the liver and lung tissues of sheep using comparative transcriptome analysis between four Tibetan sheep populations (high altitude) and one Hu sheep population (low altitude). The results showed a total of 7848 differentially expressed (DE) lncRNA transcripts, and 22,971 DE mRNA transcripts were detected by pairwise comparison. The expression patterns of selected mRNAs and lncRNAs were validated by qRT-PCR, and the results correlated well with the transcriptome data. Moreover, the functional annotation analysis based on the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases showed that DE mRNAs and the target genes of the lncRNAs were significantly enriched in organ morphogenesis, response to stimulus, haem binding, the immune system, arginine and proline metabolism, and fatty acid biosynthesis. The prediction of mRNA-mRNA and lncRNA-mRNA interaction networks further revealed transcripts potentially involved in adaptation to high-altitude hypoxia, and the hub genes DDX24, PDCD11, EIF4A3, NDUFA11, SART1, PRPF8 and TCONS_00306477, TCONS_00306029, TCONS_00139593, TCONS_00293272, and TCONS_00313398 were selected. Additionally, a set of target genes, PIK3R1, IGF1R, FZD6, IFNB2, ATF3, MB, CYP2B4, PSMD13, and TGFB1, were also identified as candidate genes associated with high-altitude hypoxia adaptation. In conclusion, a collection of novel expressed lncRNAs, a set of target genes and biological pathways known to be relevant for altitude adaptation were identified by comparative transcriptome analysis between Tibetan sheep and Hu sheep. Our results are the first to identify the characterization and expression profile of lncRNAs between Tibetan sheep and Hu sheep and provide insights into the genetic regulation mechanisms by which Tibetan sheep adapt to high-altitude hypoxic environments.


Subject(s)
Altitude Sickness , RNA, Long Noncoding , Animals , Gene Expression Profiling , Gene Regulatory Networks , Hypoxia/genetics , RNA, Long Noncoding/metabolism , RNA, Messenger/genetics , Sheep/genetics , Tibet , Transcriptome
18.
PeerJ ; 10: e13177, 2022.
Article in English | MEDLINE | ID: mdl-35368335

ABSTRACT

Bovine endometritis is an inflammatory disease of the uterus that occurs after parturition and can result in the destruction of uterine microecology, disruption of hormone secretion, and even infertility. Problems such as antibiotic residues, pathogen resistance, and microbiota dysbiosis caused by conventional antibiotic therapy cannot be ignored. According to the microecological balance theory, probiotics have the potential to prevent or cure endometritis in cattle. Probiotics can positively influence host physiology by regulating microecological imbalance, modulating immunity, and antagonizing pathogens. Since some probiotics contribute to host health only in their specific natural niches, lactic acid bacteria (LAB) from the vagina may have better potential to fight against vaginal and uterine infection. The yak (Bos grunniens) is an ancient and primitive livestock animal that is adapted to high altitude and harsh environments (cold, nutritional deficiencies, and hypoxia). However, to our knowledge, there have been no studies on yak vaginal LAB. Therefore, the purpose of this study was to isolate vaginal LAB from yak, evaluate and compare the probiotic potential and safety of the isolates, and help establish the probiotics library that can be used in the prevention and/or treatment of endometritis. Twenty-five vaginal swabs were collected from healthy yak and cultured in deMan, Rogosa, and Sharpe (MRS) broth. Tentative LAB strains were preliminarily determined through calcium dissolving zone and morphological identification, and the strains were then identified using 16S rRNA gene sequencing. The probiotics of the isolates were detected using cell aggregation, hydrophobicity, resistance to acid and bile salt, adhesion, and antibacterial activities. Additionally, antimicrobial susceptibility, hemolytic activity, and detection of potential virulence factors were determined in order to confirm the safety of these strains. Five isolates were identified: Leuconostoc mesenteroides, Lactobacillus plantarum, Enterococcus hirae, Lacticaseibacillus camelliae, and Lactobacillus mucosae. All isolates had certain growth resistance, aggregation ability, effective antimicrobial potency against Escherichia coli, Staphylococcus aureus, and Salmonella typhimurium, were sensitive to most antibiotics, and could effectively adhere to bovine endometrial epithelial cells (BEECs). None of the isolates showed hemolytic activity or harbored virulence factors. Our results indicated that the five isolates have considerable potential as probiotics that can be used to prevent and/or treat bovine endometritis. We speculate that a mixture of YD6, YD9, and YD25 may yield better results, although this would require extensive experiments to verify.


Subject(s)
Anti-Infective Agents , Endometritis , Lactobacillales , Probiotics , Female , Humans , Cattle , Animals , Lactobacillales/genetics , Endometritis/prevention & control , RNA, Ribosomal, 16S/genetics , Anti-Bacterial Agents/pharmacology , Escherichia coli/genetics , Vagina/microbiology , Probiotics/pharmacology , Virulence Factors
19.
Biochem Biophys Res Commun ; 601: 153-159, 2022 04 23.
Article in English | MEDLINE | ID: mdl-35247769

ABSTRACT

Streptococcus agalactiae mastitis is one of the significant threats to the milk industry. The traditional antibiotic treatment method is easy to cause the emergence of resistant strains, and the problem of drug residue is increasingly severe. In this study, we designed and synthesized five lipopeptides. The antibacterial activity of different molecular structure lipopeptides against Streptococcus agalactiae was detected. Furthermore, the mouse mastitis model was established using Streptococcus agalactiae. The lipopeptides with better antibacterial effect were selected for the treatment experiment to evaluate the application value in the treatment of mastitis. The results showed that 4 of the synthesized lipopeptides had specific antibacterial activity. SLP3 and SLP4 have an excellent antibacterial effect and can treat murine mastitis caused by Streptococcus agalactiae infection within the safe concentration range. The results of this study can provide an excellent experimental basis for new antibiotics and clinical application in the treatment of dairy cow mastitis.


Subject(s)
Mastitis, Bovine , Streptococcal Infections , Animals , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Cattle , Female , Humans , Lipopeptides/pharmacology , Lipopeptides/therapeutic use , Mastitis, Bovine/drug therapy , Mastitis, Bovine/microbiology , Mice , Milk/chemistry , Streptococcal Infections/drug therapy , Streptococcus agalactiae
20.
Sci Rep ; 12(1): 355, 2022 01 10.
Article in English | MEDLINE | ID: mdl-35013469

ABSTRACT

Platostoma palustre (Blume) A.J.Paton is an annual herbaceous persistent plant of the Labiatae family. However, there is a lack of genomic data for this plant, which severely restricts its genetic improvement. In this study, we performed genome survey sequencing of P. palustre and developed simple sequence repeat (SSR) markers based on the resulting sequence. K-mer analysis revealed that the assembled genome size was approximately 1.21 Gb. A total of 15,498 SSR motifs were identified and characterized in this study; among them, dinucleotide, and hexanucleotide repeats had the highest and lowest, respectively. Among the dinucleotide repeat motifs, AT/TA repeat motifs were the most abundant, and GC/CG repeat motifs were rather rare, accounting for 44.28% and 0.63%, respectively. Genetic similarity coefficient analysis by the UPMGA methods clustered 12 clones, of P. palustre and related species into two subgroups. These results provide helpful information for further research on P. palustre resources and variety improvements.


Subject(s)
DNA, Plant/genetics , Genes, Plant , Genome, Plant , High-Throughput Nucleotide Sequencing , Lamiaceae/genetics , Microsatellite Repeats , Sequence Analysis, DNA , Whole Genome Sequencing , Genetic Markers , Phylogeny
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