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1.
Materials (Basel) ; 17(11)2024 May 22.
Article in English | MEDLINE | ID: mdl-38893758

ABSTRACT

Negative thermal expansion (NTE) ceramics Sm0.85Zn0.15MnO3 (SZMO) and ZrMgMo3O12 (ZMMO) were selected to prepare Sm0.85Zn0.15MnO3-ZrMgMo3O12/Al-20Si (SZMO-ZMMO/Al-20Si) composites using ball milling and vacuum heating-press sintering processes in this study. The synergistic effect of the SZMO and ZMMO NTE ceramic reinforcements on the microstructure, mechanical properties, and coefficient of thermal expansion (CTE) of the composites was investigated. The results show that the processes of ball milling and sintering did not induce the decomposition of SZMO or ZMMO NTE ceramic reinforcements, nor did they promote a reaction between the Al-20Si matrix and SZMO or ZMMO NTE ceramic reinforcements. However, the excessive addition of SZMO and ZMMO NTE ceramics led to their aggregation within the composite. Adding a small amount of SZMO in combination with ZMMO effectively increased hardness and yield strength while reducing CTE in the Al-20Si alloy. The improvement in strength was primarily provided by SZMO, while the inhibition effect on CTE was primarily provided by ZMMO. An evaluation parameter denoted as α was proposed to evaluate the synergy effects of SZMO and ZMMO NTE ceramic reinforcements on the mechanical properties and CTE of the composites. Based on this parameter, among all composites fabricated, adding 2.5 vol% SZMO NTE ceramic and 10 vol% ZMMO NTE ceramic resulted in an optimal balance between CTE and strength for these composites with a compressive yield strength of 349.72 MPa and a CTE of 12.55 × 10-6/K, representing a significant increase in yield strength by 79.20% compared to that of Al-20Si alloy along with a notable reduction in CTE by 26.44%.

2.
Microbiol Spectr ; : e0417922, 2023 Feb 15.
Article in English | MEDLINE | ID: mdl-36790208

ABSTRACT

Legionella spp. are the causative agents of a severe pneumonia known as Legionnaires' disease. Upon being engulfed by host cells, these environmental bacteria replicate intracellularly in a plasma membrane-derived niche termed Legionella-containing vacuole (LCV) in a way that requires the defective in organelle trafficking/intracellular multiplication (Dot/Icm) protein transporter. Our understanding of interactions between Legionella and its hosts was mostly based on studies of Legionella pneumophila. In this study, we found that the LCVs created by virulent Legionella longbeachae are similarly decorated by polyubiquitinated proteins to those formed by L. pneumophila and that the ubiquitin-proteasome system (UPS) is required for optimal intracellular growth of L. longbeachae. Furthermore, we utilized bioinformatics methods and the ubiquitin-vinylmethyl ester probe to obtain potential deubiquitinases (DUBs) encoded by L. longbeachae. These efforts led to the identification of 9 L. longbeachae DUBs that displayed distinct specificity toward ubiquitin chain types. Among these, LLO_1014 and LLO_2238 are associated with the LCVs and impact the accumulation of polyubiquitinated species on the bacterial phagosome. Moreover, LLO_1014 and LLO_2238 could fully restore the phenotypes associated with Δceg23 (lotB) and Δlem27 (lotC) mutants of L. pneumophila, indicating that these DUBs have similar functions. Together, these results reveal that L. longbeachae uses multiple DUBs to construct an intracellular niche for its replication. IMPORTANCE Legionella spp. are opportunistic intracellular bacterial pathogens that cause Legionnaires' disease. Legionella utilizes the Dot/Icm type IV secretion system to deliver effector protein into host cells to modulate various cellular functions. At least 26 L. pneumophila effectors are known to hijack the host ubiquitin system via diverse mechanisms. L. longbeachae is the second leading cause of Legionnaires' disease worldwide. However, our knowledge about the interactions between L. longbeachae and its hosts is very limited. Here, we found that, similar to L. pneumophila infection, the host ubiquitin proteasome system is also important for the intracellular replication of L. longbeachae. In addition, the bacterial phagosomes harboring L. longbeachae are enriched with polyubiquitinated proteins in a Dot/Icm system-dependent manner. We further identified 9 L. longbeachae proteins that function as DUBs with distinct ubiquitin chain specificity. Of note, several of the phagosome-associated L. longbeachae DUBs regulate the recruitment of polyubiquitinated proteins to the LCV.

3.
PLoS One ; 18(2): e0281587, 2023.
Article in English | MEDLINE | ID: mdl-36758031

ABSTRACT

Legionella organisms are ubiquitous environmental bacteria that are responsible for human Legionnaires' disease, a fatal form of severe pneumonia. These bacteria replicate intracellularly in a wide spectrum of host cells within a distinct compartment termed the Legionella-containing vacuole (LCV). Effector proteins translocated by the Dot/Icm apparatus extensively modulate host cellular functions to aid in the biogenesis of the LCV and intracellular proliferation. RavZ is an L. pneumophila effector that functions as a cysteine protease to hydrolyze lipidated LC3, thereby compromising the host autophagic response to bacterial infection. In this study, we characterized the RavZ (RavZLP) ortholog in L. longbeachae (RavZLLO), the second leading cause of Legionella infections in the world. RavZLLO and RavZLP share approximately 60% sequence identity and a conserved His-Asp-Cys catalytic triad. RavZLLO is recognized by the Dot/Icm systems of both L. pneumophila and L. longbeachae. Upon translocation into the host, it suppresses autophagy signaling in cells challenged with both species, indicating the functional redundancy of RavZLLO and RavZLP. Additionally, ectopic expression of RavZLLO but not RavZLP in mammalian cells reduces the levels of cellular polyubiquitinated and polyneddylated proteins. Consistent with this process, RavZLLO regulates the accumulation of polyubiquitinated species on the LCV during L. longbeachae infection.


Subject(s)
Legionella longbeachae , Legionella pneumophila , Legionella , Legionnaires' Disease , Animals , Humans , Legionella longbeachae/metabolism , Bacterial Proteins/genetics , Legionnaires' Disease/microbiology , Vacuoles/metabolism , Ubiquitination , Phagosomes/metabolism , Autophagy , Mammals/metabolism
4.
Microbiol Spectr ; 10(6): e0294922, 2022 12 21.
Article in English | MEDLINE | ID: mdl-36377917

ABSTRACT

The increasingly serious problem of bacterial drug resistance has led to the development of antivirulence agents. The Salmonella enterica serovar Typhimurium Salmonella pathogenicity island (SPI)-encoded type III secretion system (T3SS) and its effector proteins are important virulence factors for S. Typhimurium invasion and replication in host cells and for antivirulence drug screening. Fraxetin is isolated from Fraxinus spp. Extensive studies have reported its multiple pharmacological activities. However, it remains to be elucidated whether fraxetin affects the function of the S. Typhimurium T3SS. In this study, the anti-infection mechanism of fraxetin on S. Typhimurium and its T3SS was investigated. Fraxetin inhibited the S. Typhimurium invasion of HeLa cells without affecting the growth of bacteria in vitro. Further findings on the mechanism showed that fraxetin had an inhibitory effect on the S. Typhimurium T3SS by inhibiting the transcription of the pathogenesis-related SPI-1 transcriptional activator genes hilD, hilC, and rtsA. Animal experiments showed that fraxetin treatment protected mice against S. Typhimurium infection. Collectively, we provide the first demonstration that fraxetin may serve as an effective T3SS inhibitor for the development of treatments for Salmonella infection. IMPORTANCE The increasingly serious problem of bacterial antibiotic resistance limits the clinical application of antibiotics, which increases the need for the development of antivirulence agents. The type III secretion system (T3SS) plays a critical role in host cell invasion and pathogenesis of Salmonella and becomes a popular target for antivirulence agents screening. Our study found, for the first time, that fraxetin inhibited S. Typhimurium invasion by inhibiting the transcription of genes in a feed-forward regulatory loop. Further in vivo testing showed that fraxetin decreased bacterial burdens in the spleen and liver of S. Typhimurium-infected mice and improved survival outcomes in an in vivo mouse model of S. Typhimurium infection. Collectively, these results demonstrate that fraxetin inhibits S. Typhimurium infection by targeting the T3SS and may serve as a potential agent for the treatment of S. Typhimurium infection.


Subject(s)
Salmonella typhimurium , Type III Secretion Systems , Humans , Animals , Mice , Type III Secretion Systems/metabolism , HeLa Cells , Serogroup , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial
5.
Front Cell Infect Microbiol ; 12: 967149, 2022.
Article in English | MEDLINE | ID: mdl-36176578

ABSTRACT

New therapeutic strategies for clinical Salmonella enterica serovar Typhimurium (S. Typhimurium) infection are urgently needed due to the generation of antibiotic-resistant bacteria. Inhibition of bacterial virulence has been increasingly regarded as a potential and innovative strategy for the development of anti-infection drugs. Salmonella pathogenicity island (SPI)-encoded type III secretion system (T3SS) represents a key virulence factor in S. Typhimurium, and active invasion and replication in host cells is facilitated by the secretion of T3SS effector proteins. In this study, we found that harmine could inhibit T3SS secretion; thus, its potential anti-S. Typhimurium infection activity was elucidated. Harmine inhibits the secretion and expression of T3SS effector proteins and consequently attenuates the S. Typhimurium invasion function of HeLa cells. This inhibition may be implemented by reducing the transcription of pathogenesis-related SPI-1 transcriptional activator genes hilD, hilC, and rtsA. Harmine improves the survival rate and bacterial loads of mice infected with S. Typhimurium. In summary, harmine, an effective T3SS inhibitor, could be a leading compound for the development of treatments for Salmonella infection.


Subject(s)
Salmonella typhimurium , Type III Secretion Systems , Animals , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial , Harmine/metabolism , Harmine/pharmacology , HeLa Cells , Humans , Mice , Salmonella typhimurium/genetics , Serogroup , Type III Secretion Systems/metabolism , Virulence Factors/metabolism
6.
Micron ; 154: 103202, 2022 Mar.
Article in English | MEDLINE | ID: mdl-34998138

ABSTRACT

Al-5.8Mg-0.4Mn with minor alloying additions of Sc and Zr was investigated via electrochemical testing and nitric acid mass loss testing (NAMLT) in order to reveal the influence of Sc and Zr upon sensitization and intergranular corrosion. The Al-Mg-Mn alloys were also analysed using an electron probe microanalyzer, indicating that ß-phase (Al3Mg2) was more likely to precipitate around rectangular or cubic Al6Mn particles. Results reveal that the strength and intergranular corrosion resistance of Al-5.8Mg-0.4Mn was improved by the combined addition of Sc and Zr. The Al3(Sc1-xZrx) dispersoids can lead to an alteration of the relative proportion of low angle grain boundaries, and lower volume fraction of ß-phase was observed for Al-5.8Mg-0.4Mn-xSc-yZr relative to the Sc and Zr free alloy.

7.
Micron ; 147: 103082, 2021 Aug.
Article in English | MEDLINE | ID: mdl-34022758

ABSTRACT

Pre-alloyed CoCrFeMnNi high-entropy alloy (HEA) powders were prepared by gas atomization and consequently hot pressing sintered (HPS) at 1100 °C for 2 h for fabrication bulk materials. Sintered equiatomic CoCrFeMnNi HEA exhibited a homogeneous FCC structured single-phase solid solution and equiaxed grains with an average size of ∼16 µm. Analyses using TEM showed that sheet-like metastable structures with a size of ∼55 to 160 nm were formed in the sintered bulks. The nano-sized metastable structures were inherited from the gas atomized CoCrFeMnNi powders with nano-sized crystallites formed during the rapid solidification process. The room temperature yield and ultimate tensile strength of sintered HEA reached 358 and 778 MPa, respectively, and the alloy kept excellent plasticity of ∼28 %. Investigations of the deformation substructures at specific strain levels with EBSD revealed the sintered CoCrFeMnNi HEA kept FCC structured single phase and the main deformation mechanism was dislocation slip. The strengthening mechanism can be attributed to the combination effects of grain refinement and the presence of nano-sized metastable structures.

8.
Front Vet Sci ; 7: 294, 2020.
Article in English | MEDLINE | ID: mdl-32548131

ABSTRACT

Canine circovirus (CanineCV), a new pathogen, was found to be associated with canine hemorrhagic diarrhea, vasculitis, granulomatous lymphadenitis, and acute gastroenteritis. Although CanineCV was highly positive rate in diarrhea cases, its pathogenicity remains controversial. In this study, the seroprevalence and associated risk factors of CanineCV infection among domestic dogs in northeastern China was investigated by an indirect enzyme-linked immunosorbent assay (iELISA) based on recombinant capsid protein. Results revealed the proposed iELISA had no cross-reactivity with other related pathogens, and yielded good diagnostic values. Then, to evaluate the rCap iELISA, this study applied it to detect antibodies against CanineCV in 1,047 clinical serum samples obtained from northeastern China in 2016-2017. Results showed the positive rates in the five cities of Jilin, Liaoning, and Heilongjiang provinces ranged from 22.22 to 42.29%. Statistical analysis shows a significant difference in age between dogs <3 months old with respect to the >1-year-old dogs (p = 0.005), that is, the CanineCV infection was more frequently identified from older dogs. In the artificially infected experiment, the dogs developed seroconversion after 9 or 12 days and the main way of virus excretion was through feces. More interestingly, among the 32 ELISA-positive serum samples, 34.75% samples tested positive for the CanineCV DNA by qPCR, far higher than that in ELISA-negative serum samples (5.26%, 2/38). This report is the first to demonstrate that CanineCV infection is common in the dog population in northeastern China. The results showed obvious differences in the positive rate associated with diarrhea, age, but not with different cities. This study also provide basis for evaluating the pathogenic potential of CanineCV. But, the pathogenicity, the relationship between antibody level and immune protection, and the harmful effects of this virus remain to be established.

9.
Article in English | MEDLINE | ID: mdl-30736004

ABSTRACT

Identifying protein complexes is helpful for understanding cellular functions and designing drugs. In the last decades, many computational methods have been proposed based on detecting dense subgraphs or subnetworks in Protein-Protein Interaction Networks (PINs). However, the high rate of false positive/negative interactions in PINs prevents from the achievement of satisfactory detection results directly from PINs, because most of such existing methods exploit mainly topological information to do network partitioning. In this paper, we propose a new approach for protein complex detection by merging topological information of PINs and functional information of proteins. We first split proteins to a number of protein groups from the perspective of protein functions by using FunCat data. Then, for each of the resulting protein groups, we calculate two protein-protein similarity matrices: one is computed by using graph embedding over a PIN, the other is by using GO terms, and combine these two matrices to get an integrated similarity matrix. Following that, we cluster the proteins in each group based on the corresponding integrated similarity matrix, and obtain a number of small protein clusters. We map these clusters of proteins onto the PIN, and get a number of connected subgraphs. After a round of merging of overlapping subgraphs, finally we get the detected complexes. We conduct empirical evaluation on four PPI datasets (Collins, Gavin, Krogan, and Wiphi) with two complex benchmarks (CYC2008 and MIPS). Experimental results show that our method performs better than the state-of-the-art methods.


Subject(s)
Computational Biology/methods , Protein Interaction Mapping/methods , Protein Interaction Maps/genetics , Algorithms , Cluster Analysis , Databases, Protein , Transcriptome/genetics
10.
Arch Virol ; 165(1): 137-143, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31745718

ABSTRACT

Canine circovirus (canine CV) is an etiological agent associated with diarrhea, hemorrhagic gastroenteritis and vasculitis. Although canine CV has been identified and characterized in southern China in recent years, its epidemiology in other regions of China and its precise molecular characteristics have not been examined. In this study, we examined 141 fecal specimens collected from domestic dogs with or without diarrhea in Heilongjiang province, Northeastern China, during 2014 to 2016. A total of 18 out of 141 samples were found to be positive for canine CV by real-time quantitative PCR. In the diarrhea samples, canine CV was detected in coinfections with canine parvovirus 2. More importantly, two different canine CV strains were detected in one sample. Five canine CV genomes were successfully amplified. Sequence analysis showed that there were two unique amino acid changes in the Rep protein (N39S in the K1 strain, and T71A in the XF16 strain). Phylogenetic analysis indicated that canine CV could be divided into four genotypes, and specific nucleotide mutations could be used for confirming the four genotypes. Moreover, recombination analysis revealed that a total of eight recombination events were found in five genomic sequences. Molecular evolution analysis showed that the canine CV has been under purifying selection. This study provides evidence that at least three genotypes of canine CV are co-circulating in China. Continuous epidemiological surveillance is therefore necessary to understand their importance for the evolution of canine CV.


Subject(s)
Circovirus/classification , Diarrhea/veterinary , Dog Diseases/virology , Mutation , Parvovirus, Canine/isolation & purification , Animals , China , Circovirus/genetics , Diarrhea/virology , Dogs , Evolution, Molecular , Feces/virology , Parvovirus, Canine/genetics , Phylogeny , Viral Proteins/genetics , Whole Genome Sequencing
11.
Microb Pathog ; 127: 12-20, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30496836

ABSTRACT

Biosurfactants (BS) are amphipathic compounds produced by diverse groups of microorganisms exhibiting various biological activities. The current study aimed to assess antimicrobial, anti-adhesive and anti-biofilm activities of BS isolated from lactic acid bacteria (LAB), including Pediococcus acidilactici and Lactobacillus plantarum against Staphylococcus aureus CMCC 26003 in vitro. Cell-bound BS from both Pediococcus acidilactici and Lactobacillus plantarum were extracted, and their surface activities were evaluated by oil spreading assay. As quantified by crystal violet method, BS inhibited adhesion and biofilm formation of Staphylococcus aureus in a dose-dependent manner. The above findings were further supported by results of scanning electron microscopy. These two kinds of BS affect expressions of biofilm-related genes (cidA, icaA, dltB, agrA, sortaseA and sarA) and interfere with the release of signaling molecules (AI-2) in quorum sensing systems. Biological activities observed for BS produced by tested LAB suggest prospects for their use against Staphylococcus aureus biofilm-related infections.


Subject(s)
Anti-Infective Agents/pharmacology , Bacterial Adhesion/drug effects , Biofilms/drug effects , Lactobacillus plantarum/chemistry , Pediococcus acidilactici/chemistry , Staphylococcus aureus/drug effects , Surface-Active Agents/pharmacology , Anti-Infective Agents/isolation & purification , Bacterial Proteins/biosynthesis , Biofilms/growth & development , Biological Products/isolation & purification , Biological Products/pharmacology , Gene Expression Profiling , Gentian Violet/analysis , Microscopy, Electron, Scanning , Quorum Sensing/drug effects , Staining and Labeling , Staphylococcus aureus/physiology , Surface-Active Agents/isolation & purification
12.
Probiotics Antimicrob Proteins ; 11(4): 1086-1099, 2019 12.
Article in English | MEDLINE | ID: mdl-30056601

ABSTRACT

The antioxidant activities and probiotic properties of lactic acid bacteria (LAB) selected from traditional artisanal milk cheese from Northeast China were investigated in this study. Among the 322 isolates, 175 LAB were identified through probiotic characterizations. Twenty-three out of the 175 strains exhibited antibacterial activity against more than four enteropathogenic bacteria. The antioxidant action of 23 LAB was evaluated by different methods, including scavenging of hydroxide radicals, DPPH radicals, superoxide anions, and ABTS+ radical cation. The ability to resist hydrogen peroxide and superoxide dismutase activity was also studied. These strains significantly showed antioxidative capacity compared with a non-antioxidative strain, closely followed by the standard probiotic strain Lactobacillus rhamnosus GG or even better. Based on 16S ribosomal RNA-sequence analysis, the 23 isolates belonged to the species Lactobacillus plantarum (16), Lactobacillus paracasei (2), Enterococcus faecium (2), Lactobacillus helveticus (1), Weissella paramesenteroides (1), and Pediococcus pentosaceus (1). In addition, five out of the 23 strains were susceptible to most of the tested antibiotics, showed extremely high levels of hydrophobicity similar to or better than the reference strain L. rhamnosus GG, and did not exhibit any hemolytic activity. These five strains were also confirmed safe through bacterial translocation. Results suggest that at least five probiotic candidates can be explored as prospective antioxidants and used as a potential antioxidant strain to be utilized in the development of functional foods and new starter cultures.


Subject(s)
Antioxidants/pharmacology , Cheese/microbiology , Lactobacillales/isolation & purification , Probiotics/pharmacology , Animals , Antibiosis , Antioxidants/chemistry , China , Hydrophobic and Hydrophilic Interactions , Lactobacillales/classification , Lactobacillales/genetics , Lactobacillales/physiology , Milk/microbiology , Probiotics/chemistry , Probiotics/classification
13.
Arch Virol ; 163(10): 2727-2735, 2018 Oct.
Article in English | MEDLINE | ID: mdl-29948383

ABSTRACT

Mink circovirus (MiCV), a virus that was newly discovered in 2013, has been associated with enteric disease. However, its etiological role in acute gastroenteritis is unclear, and its genetic characteristics are poorly described. In this study, the role of circoviruses (CVs) in mink acute gastroenteritis was investigated, and the MiCV genome was molecularly characterized through sequence analysis. Detection results demonstrated that MiCV was the only pathogen found in this infection. MiCVs and previously characterized CVs shared genome organizational features, including the presence of (i) a potential stem-loop/nonanucleotide motif that is considered to be the origin of virus DNA replication; (ii) two major inversely arranged open reading frames encoding putative replication-associated proteins (Rep) and a capsid protein; (iii) direct and inverse repeated sequences within the putative 5' region; and (iv) motifs in Rep. Pairwise comparisons showed that the capsid proteins of MiCV shared the highest amino acid sequence identity with those of porcine CV (PCV) 2 (45.4%) and bat CV (BatCV) 1 (45.4%). The amino acid sequence identity levels of Rep shared by MiCV with BatCV 1 (79.7%) and dog CV (dogCV) (54.5%) were broadly similar to those with starling CV (51.1%) and PCVs (46.5%). Phylogenetic analysis indicated that MiCVs were more closely related to mammalian CVs, such as BatCV, PCV, and dogCV, than to other animal CVs. Among mammalian CVs, MiCV and BatCV 1 were the most closely related. This study could contribute to understanding the potential pathogenicity of MiCV and the evolutionary and pathogenic characteristics of mammalian CVs.


Subject(s)
Circoviridae Infections/veterinary , Circovirus/genetics , Mink/virology , Animals , Capsid Proteins/genetics , Capsid Proteins/metabolism , China , Circoviridae Infections/virology , Circovirus/classification , Circovirus/isolation & purification , Gastroenteritis/virology , Genome, Viral , Genomics , Open Reading Frames , Phylogeny
14.
Front Microbiol ; 9: 937, 2018.
Article in English | MEDLINE | ID: mdl-29867846

ABSTRACT

The mink circovirus (MiCV), a newly discovered pathogen, is associated with diarrhea in farmed minks. The prevalence and economic importance of this virus remain poorly understood, and a quantitative method for diagnosis of MiCV infection has not been established. This research aims to develop a highly specific, sensitive, and quantitative assay for MiCV. A Real-Time quantitative polymerase chain reaction (qPCR) assay was developed to detect different isolates of the MiCV in mink samples. The qPCR system is highly sensitive with a detection limit of as low as 10 viral DNA copies. The specificity of this qPCR assay was supported by the absence of cross-reaction with other pathogens. The coefficients of variation were low for both inter-assay and intra-assay variabilities. In addition, the results also expressed the distribution of MiCV in infectious mink tissues with high levels of virus in the skeletal muscle and heart. The heart occupied a higher proportion than other tissues, which can be considered the primary source of test material. This qPCR method could be a useful tool for epidemiological studies and disease management. This method for MiCV is highly specific, sensitive, repeatable, quantitative, and can rapidly determine viral load levels in different tissues samples.

15.
J Virol Methods ; 256: 1-5, 2018 06.
Article in English | MEDLINE | ID: mdl-29501532

ABSTRACT

To date, the pathogenic role of mink circovirus (MiCV) remains unclear, and its prevalence and economic importance are unknown. Therefore, a rapid and sensitive molecular diagnosis is necessary for disease management and epidemiological surveillance. However, only PCR methods can identify MiCV infection at present. In this study, we developed a nested PCR and established a novel recombinase polymerase amplification (RPA) assay for MiCV detection. Sensitivity analysis showed that the detection limit of nested PCR and RPA assay was 101 copies/reaction, and these methods were more sensitive than conventional PCR, which has a detection limit of 105 copies/reaction. The RPA assay had no cross-reactivity with other related viral pathogens, and amplification was completed in less than 20 min with a simple device. Further assessment of clinical samples showed that the two assays were accurate in identifying positive and negative conventional PCR samples. The detection rate of MiCV by the RPA assay in clinical samples was 38.09%, which was 97% consistent with that by the nested PCR. The developed nested PCR is a highly sensitive tool for practical use, and the RPA assay is a simple, sensitive, and potential alternative method for rapid and accurate MiCV diagnosis.


Subject(s)
Animal Diseases/diagnosis , Animal Diseases/virology , Circoviridae Infections/veterinary , Circovirus/genetics , Mink , Nucleic Acid Amplification Techniques , Animals , Polymerase Chain Reaction , Sensitivity and Specificity
16.
BMC Vet Res ; 14(1): 29, 2018 Jan 26.
Article in English | MEDLINE | ID: mdl-29373971

ABSTRACT

BACKGROUND: Mink circovirus (MiCV) is a newly discovered pathogen associated with mink diarrhea. The prevalence and economic importance of this virus remain poorly understood, and no specific serological assay has been developed for the diagnosis of MiCV infection. RESULTS: In this study, a recombinant capsid protein antigen expressed in Escherichia coli was utilized to establish an indirect enzyme-linked immunosorbent assay (iELISA). Results revealed that the assay had no cross-reactivity with other related pathogens, and the respective sensitivity and specificity of the proposed iELISA were 92.31% and 91.67% compared with those obtained of Western blot on 138 serum samples from minks. The correlation coefficient between iELISA and Western blot was 0.838 (p > 0.05). iELISA was applied to detect MiCV antibodies in 683 clinical serum samples from different farms from the major mink industry province in China, and 21 of 24 farms with 163 of 683 (23.87%) individuals were tested positive for MiCV antibodies. The positive rates of each of the 21 flocks ranged from 2.33% to 73.68%. CONCLUSIONS: These results indicated that iELISA was a sensitive and specific method suitable for the large-scale detection of MiCV infections in mink. This study provided an effective method for the serological diagnosis and positive rate investigation of MiCV infection.


Subject(s)
Circoviridae Infections/veterinary , Circovirus , Enzyme-Linked Immunosorbent Assay/veterinary , Mink/virology , Animals , Capsid Proteins/immunology , Circoviridae Infections/immunology , Circoviridae Infections/virology , Enzyme-Linked Immunosorbent Assay/methods , Recombinant Proteins/immunology , Reproducibility of Results , Sensitivity and Specificity , Serologic Tests/veterinary
17.
Probiotics Antimicrob Proteins ; 10(4): 601-610, 2018 12.
Article in English | MEDLINE | ID: mdl-29218505

ABSTRACT

The present study aims to investigate the probiotic properties of novel strains of lactic acid bacteria isolated from traditional artisanal milk cheese from Northeast China and to explore their antibacterial activity against enteropathogenic bacteria. Of the 321 isolates, 86 exhibited survival in low pH, resistance to pancreatin, and tolerance to bile salts; of these, 12 inhibited the growth of more than seven enteropathogenic bacteria and exhibited antibiofilm activities against Staphylococcus aureus CMCC26003 and/or Escherichia coli CVCC230. Based on 16S ribosomal RNA sequence analysis, the 12 isolates were assigned to Lactobacillus plantarum (7), Lactobacillus helveticus (3), Pediococcus acidilactici (1), and Enterococcus faecium (1) species. In addition, 5 of the 12 strains were susceptible to most of the tested antibiotics. Furthermore, four strains with sensitivity to antibiotics showed significantly high levels of hydrophobicity similar to or better than the reference strain Lactobacillus rhamnosus GG. Moreover, three strains were confirmed safe through non-hemolytic activities and bacterial translocation. Overall, the selected Lact. plantarum 27053 and 27172 and Lact. helveticus 27058 strains can be considered potential probiotic strains and candidates for further application in functional food and prevention or treatment of gastrointestinal diseases.


Subject(s)
Antibiosis , Biofilms , Cheese/microbiology , Escherichia coli/physiology , Lactobacillales/isolation & purification , Milk/microbiology , Staphylococcus aureus/physiology , Animals , Cattle , China , Escherichia coli Infections/microbiology , Lactobacillales/classification , Lactobacillales/genetics , Lactobacillales/physiology , Mice , Probiotics/isolation & purification , Staphylococcal Infections/microbiology
18.
Arch Microbiol ; 199(9): 1267-1275, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28616631

ABSTRACT

Staphylococcus aureus (S. aureus) is a Gram-positive pathogen and forms biofilm easily. Bacteria inside biofilms display an increased resistance to antibiotics and disinfectants. The objective of the current study was to assess the antimicrobial activities of emodin, 1,2,8-trihydroxy-6-methylanthraquinone, an anthraquinone derivative isolated from Polygonum cuspidatum and Rheum palmatum, against S. aureus CMCC26003 grown in planktonic and biofilm cultures in vitro. In addition, a possible synergistic effect between emodin and berberine chloride was evaluated. As quantified by crystal violet method, emodin significantly decreased S. aureus biofilm growth in a dose-dependent manner. The above findings were further supported by scanning electron microscopy. Moreover, the present study demonstrated that sub-MICs emodin obviously intervened the release of extracellular DNA and inhibited expression of the biofilm-related genes (cidA, icaA, dltB, agrA, sortaseA and sarA) by real-time RT-PCR. These results revealed a promising application for emodin as a therapeutic agent and an effective strategy to prevent S. aureus biofilm-related infections.


Subject(s)
Anti-Bacterial Agents/pharmacology , Berberine/pharmacology , Biofilms/drug effects , Emodin/pharmacology , Staphylococcus aureus/drug effects , Biofilms/growth & development , Drug Synergism , Gene Expression/drug effects , Microbial Sensitivity Tests , Microscopy, Electron, Scanning , Plankton/drug effects , Plankton/microbiology , Staphylococcus aureus/genetics
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