ABSTRACT
Infrared target detection is widely used in industrial fields, such as environmental monitoring, automatic driving, etc., and the detection of weak targets is one of the most challenging research topics in this field. Due to the small size of these targets, limited information and less surrounding contextual information, it increases the difficulty of target detection and recognition. To address these issues, this paper proposes YOLO-ISTD, an improved method for infrared small target detection based on the YOLOv5-S framework. Firstly, we propose a feature extraction module called SACSP, which incorporates the Shuffle Attention mechanism and makes certain adjustments to the CSP structure, enhancing the feature extraction capability and improving the performance of the detector. Secondly, we introduce a feature fusion module called NL-SPPF. By introducing an NL-Block, the network is able to capture richer long-range features, better capturing the correlation between background information and targets, thereby enhancing the detection capability for small targets. Lastly, we propose a modified K-means clustering algorithm based on Distance-IoU (DIoU), called K-means_DIOU, to improve the accuracy of clustering and generate anchors suitable for the task. Additionally, modifications are made to the detection heads in YOLOv5-S. The original 8, 16, and 32 times downsampling detection heads are replaced with 4, 8, and 16 times downsampling detection heads, capturing more informative coarse-grained features. This enables better understanding of the overall characteristics and structure of the targets, resulting in improved representation and localization of small targets. Experimental results demonstrate significant achievements of YOLO-ISTD on the NUST-SIRST dataset, with an improvement of 8.568% in mAP@0.5 and 8.618% in mAP@0.95. Compared to the comparative models, the proposed approach effectively addresses issues of missed detections and false alarms in the detection results, leading to substantial improvements in precision, recall, and model convergence speed.
Subject(s)
Algorithms , Infrared Rays , Cluster Analysis , Pattern Recognition, Automated/methodsABSTRACT
The variation of solid Earth's hydrologic loading could cause the elastic vertical deformation of the crust, and the Global Navigation Satellite System (GNSS) could effectively monitor the vertical displacement of surface loads. However, the widely used Green's function method does not work well in areas where GNSS sites are sparse. Here, the vertical displacement time series of GNSS stations and the Slepian basis function method have been applied to convert displacement signals into spatial spectrum signals. The elastic mass load theory is used to study the changes in terrestrial water storage on the Northeastern Tibetan Plateau (NETP). The temporal and spatial characteristics of seasonal water changes are well-represented by the GNSS, the Gravity Recovery and Climate Experiment (GRACE), and the Global Land Data Assimilation System (GLDAS). Several data points suggest that the change in water storage shows a gradual increase from the northeast to the southwest. The greatest annual amplitude of water storage retrieved by GNSS is â¼159 mm, which is greater than the â¼47 mm and â¼44 mm obtained by GRACE and GLDAS. These results demonstrate that GNSS is capable of capturing small-scale hydrological changes in this region, whereas GRACE and GLDAS data tend to underestimate seasonal variations in water storage. We also used GNSS to describe the hydrological drought conditions in NETP, showing that GNSS could be used as an independent method to characterize hydrological drought events. The findings suggest it could observe water storage with high spatial and temporal resolution and aid in comprehending regional hydrological trends with a sparse GNSS station network.
ABSTRACT
Streptococcus suis serotype 2 (SS2) has been reported to be a highly invasive pathogen in swine and a zoonotic agent for humans. Although many bacterial virulence factors have been identified, our an insightful understanding of SS2 pathogenicity is lacking. The gene nadR, encoding nicotinamide-nucleotide adenylyltransferase, was first reported as a regulator and transporter of the intracellular NAD synthesis pathway in Salmonella typhimurium. In this study, we constructed a mutant strain of nadR (ΔnadR) to test the phenotypic and virulence variations between the deletion mutant and the wild-type strain ZY05719. The phenotypic experimental results showed that ΔnadR obviously inhibited cell growth and exhibited shorter chains than WT. The growth defect of ΔnadR was caused by the loss of the function of nadR for transporting the substrates nicotinamide mononucleotide and nicotinamide riboside in the intracellular NAD synthesis pathway. In the process of interaction with the host, ΔnadR participated in adherence and invasion to the host cells, and it was more easily cleared by RAW264.7 macrophages. More importantly, both zebrafish and BALB/c mice in vivo virulence experimental results showed that ΔnadR dramatically attenuated the virulence of SS2, and the ability of ΔnadR to colonize tissues was notably reduced in comparison with that of WT in the BALB/c mice infection model. To the best of our knowledge, this is the first time to demonstrate that nadR not only plays an important role in bacterial growth, but also in connection with the virulence of SS2 as a global transcriptional regulator.
Subject(s)
Streptococcal Infections , Streptococcus suis , Swine Diseases , Animals , Mice , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , NAD/metabolism , Serogroup , Streptococcal Infections/veterinary , Streptococcal Infections/microbiology , Streptococcus suis/pathogenicity , Swine , Virulence/genetics , ZebrafishABSTRACT
IMPORTANCE: To the best of our knowledge, this study reveals a strong correlation between mass spectra pattern and virulence phenotype among S. suis for the first time. In order to make the findings applicable and to excavate the intrinsic information in the spectra, the classifiers based on the machine learning algorithms were established, and RF (Random Forest)-based models have achieved an accuracy of over 90%. Overall, this study will pave the way for virulent SS2 (Streptococcus suis serotype 2) rapid detection, and the important findings on the association between genotype and mass spectrum may provide a new idea for the genotype-dependent detection of specific pathogens.
Subject(s)
Streptococcal Infections , Streptococcus suis , Humans , Virulence/genetics , Streptococcus suis/genetics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Serogroup , Machine LearningABSTRACT
Infectious serositis of ducks, caused by Riemerella anatipestifer, is one of the main infectious diseases that harm commercial ducks. Whole-strain-based vaccines with no or few cross-protection were observed between different serotypes of R. anatipestifer, and so far, control of infection is hampered by a lack of effective vaccines, especially subunit vaccines with cross-protection. Since the concept of reverse vaccinology was introduced, it has been widely used to screen for protective antigens in important pathogens. In this study, pan-genome binding reverse vaccinology, an emerging approach to vaccine candidate screening, was used to screen for cross-protective antigens against R. anatipestifer. Thirty proteins were identified from the core-genome as potential cross-protective antigens. Three of these proteins were recombinantly expressed, and their immunoreactivity with five antisera (anti-serotypes 1, 2, 6, 10, and 11) was demonstrated by Western blotting. Our study established a method for high-throughput screening of cross-protective antigens against R. anatipestifer in silico, which will lay the foundation for the development of a cross-protective subunit vaccine controlling R. anatipestifer infection. KEY POINTS: ⢠Pan-genome binding reverse vaccine approach was first established in R. anatipestifer to screen for subunit vaccine candidates. ⢠Thirty potential cross-protective antigens against R. anatipestifer were identified by this method. ⢠The reliability of the method was verified preliminarily by the results of Western blotting of three of these potential antigens.
Subject(s)
Flavobacteriaceae Infections , Poultry Diseases , Riemerella , Animals , Poultry Diseases/prevention & control , Reproducibility of Results , Riemerella/genetics , Vaccines, Subunit , Ducks , Flavobacteriaceae Infections/prevention & control , Flavobacteriaceae Infections/veterinaryABSTRACT
AIM: Combining MALDI-TOF MS and machine learning to establish a new rapid method to identify two important serotypes of Rimerella anatipestifer. METHODS AND RESULTS: MALDI-TOF MS was performed on 115 R. anatipestifer strains (serotype 1, serotype 2, and other serotypes) to explore its ability to identify serotypes of R. anatipestifer. Raw spectral data were generated in diagnostic mode; these data were preprocessed, clustered, and analysed using principal component analysis. The results indicated that MALDI-TOF MS completely differentiated serotype 1 from serotype 2 of R. anatipestifer; the potential serotype-associated m/z loci are listed. Furthermore, Random Forest and Support Vector Machine were used for modelling to identify the two important serotypes, and the results of cross-validation indicated that they had â¼80% confidence to make the right classification. CONCLUSION: We proved that MALDI-TOF MS can differentiate serotype 1 from serotype 2 of R. anatipestifer. Additionally, the identification models established in this study have high confidence to screen out these two important serotypes from other serotypes.
Subject(s)
Poultry Diseases , Riemerella , Animals , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Serogroup , Riemerella/genetics , Birds , Machine LearningABSTRACT
Avian pathogenic Escherichia coli, a causative agent of avian colibacillosis, has been causing serious economic losses in the poultry industry. The increase in multidrug-resistant isolates and the complexity of the serotypes of this pathogen, especially the recently reported emergence of a newly predominant serogroup of O145, make the control of this disease difficult. To address this challenge, a high-throughput screening approach, called Pan-RV (Reverse vaccinology based on pangenome analysis), is proposed to search for universal protective antigens against the three traditional serogroups and the newly emerged O145. Using this approach, a total of 61 proteins regarded as probable antigens against the four important serogroups were screened from the core genome of 127 Avian pathogenic Escherichia coli (APEC) genomes, and six were verified by Western blots using antisera. Overall, our research will provide a foundation for the development of an APEC subunit vaccine against avian colibacillosis. Given the exponential growth of whole-genome sequencing (WGS) data, our Pan-RV pipeline will make screening of bacterial vaccine candidates inexpensive, rapid, and efficient. IMPORTANCE With the emergence of drug resistance and the newly predominant serogroup O145, the control of Avian pathogenic Escherichia coli is facing a serious challenge; an efficient immunological method is urgently needed. Here, for the first time, we propose a high-throughput screening approach to search for universal protective antigens against the three traditional serogroups and the newly emerged O145. Importantly, using this approach, a total of 61 proteins regarded as probable antigens against the four important serogroups were screened, and three were shown to be immunoreactive with all antisera (covering the four serogroups), thereby providing a foundation for the development of APEC subunit vaccines against avian colibacillosis. Further, our Pan-RV pipeline will provide immunological control strategies for pathogens with complex and variable genetic backgrounds such as Escherichia coli and will make screening of bacterial vaccine candidates more inexpensive, rapid, and efficient.
Subject(s)
Escherichia coli Infections , Escherichia coli Vaccines , Poultry Diseases , Animals , Escherichia coli/genetics , Serogroup , Escherichia coli Infections/prevention & control , Escherichia coli Infections/veterinary , Poultry , Bacterial Vaccines , Poultry Diseases/prevention & control , Poultry Diseases/microbiology , ChickensABSTRACT
OBJECTIVES: Polymyxin is considered as one of the 'last lines of defense' for the treatment of multidrug resistant bacteria. Increased use of polymyxin during recent years poses a risk to public health. The purpose of this study was to investigate the carrying situation of the mcr-1 drug-resistance gene in waterfowl in some coastal areas of China from 2019 to 2020. METHODS: Fifty-seven isolated avian pathogenic Escherichia coli strains were selected from 493 APEC isolates for whole-genome sequencing. The 24 mcr-1-positive APEC strains were tested for conjugation and genome-wide analysis, including sequence type (ST) analysis, serotype analysis, and drug-resistance gene analysis. Numerous mcr-1-positive E. coli were downloaded from the National Center for Biotechnology Information (NCBI) for comparative genomic analysis. RESULTS: Antimicrobial susceptibility test results showed that 57 APEC isolates were highly resistant to gentamicin, cefotaxime, and ofloxacin, and 24 mcr-1-positive APEC isolates were resistant to polymyxin. Fourteen isolates of mcr-1-positive APEC plasmids were successfully conjugated to EC600. Both ST156 and ST10 were found in high proportions in human and avian sources through genome-wide analysis; it is worth noting that these two isolates of APEC were detected to contain the blaNDM-1 and blaNDM-4 genes, respectively. CONCLUSION: In this study, the epidemiological investigation of the mcr-1 gene was carried out on APEC in some coastal areas of China from 2019 to 2020, and our results have enriched the data on the transmission of APEC isolates carrying the mcr-1 gene in waterfowl.
Subject(s)
Escherichia coli Proteins , Escherichia coli , Anti-Bacterial Agents/pharmacology , Colistin , Drug Resistance, Bacterial/genetics , Escherichia coli Proteins/genetics , Humans , PolymyxinsABSTRACT
Klebsiella pneumoniae is an opportunistic Gram-negative bacterium that has become a leading causative agent of nosocomial infections, mainly infecting patients with immunosuppressive diseases. Capsular (K) serotypes K1, K2, K47, and K64 are commonly associated with higher virulence (hypervirulent Klebsiella pneumoniae), and more threateningly, isolates belonging to the last two K serotypes are also frequently associated with resistance to carbapenem (hypervirulent carbapenem-resistant Klebsiella pneumoniae). The prevalence of these isolates has posed significant threats to human health, and there are no appropriate therapies available against them. Therefore, in this study, a method combining immunoinformatics and pangenome analysis was applied for contriving a multiepitope subunit vaccine against these four threatening serotypes. To obtain cross-protection, 12 predicted conserved antigens were screened from the core genome of 274 complete Klebsiella pneumoniae genomes (KL1, KL2, KL47, and KL64), from which the epitopes of T and B cells were extracted for vaccine construction. In addition, the immunological properties, the interaction with Toll-like receptors, and the stability in a simulative humoral environment were evaluated by immunoinformatics methods, molecular docking, and molecular dynamics simulation. All of these evaluations indicated the potency of this constructed vaccine to be an effective therapeutic agent. Lastly, the cDNA of the designed vaccine was optimized and ligated to pET-28a(+) for expression vector construction. Overall, our research provides a newly cross-protective control strategy against these troublesome pathogens and paves the way for the development of a safe and effective vaccine. IMPORTANCE Klebsiella pneumoniae is an opportunistic Gram-negative bacterium that has become a leading causative agent of nosocomial infections. Among the numerous capsular serotypes, K1, K2, K47, and K64 are commonly associated with higher virulence (hypervirulent K. pneumoniae). More threateningly, the last two serotypes are frequently associated with resistance to carbapenem (hypervirulent carbapenem-resistant K. pneumoniae). However, there is currently no therapeutic agent or vaccine specifically against these isolates. Therefore, development of a vaccine against these pathogens is very essential. In this study, for the first time, a method combining pangenome analysis, reverse vaccinology, and immunoinformatics was applied for contriving a multiepitope subunit vaccine against K. pneumoniae isolates of K1, K2, K47, and K64. Also, the immunological properties of the constructed vaccine were evaluated and its high potency was revealed. Overall, our research will pave the way for the vaccine development against these four threatening capsular serotypes of K. pneumoniae.
Subject(s)
Cross Infection , Klebsiella Infections , Anti-Bacterial Agents/therapeutic use , Carbapenems/therapeutic use , Cross Infection/drug therapy , Humans , Klebsiella Infections/drug therapy , Klebsiella Infections/epidemiology , Klebsiella Infections/prevention & control , Klebsiella pneumoniae/genetics , Molecular Docking Simulation , Vaccines, Subunit/genetics , Vaccines, Subunit/therapeutic useABSTRACT
Streptococcus parasuis (S. parasuis) is a close relative of Streptococcus suis (S. suis), composed of former members of S. suis serotypes 20, 22 and 26. S. parasuis could infect pigs and cows, and recently, human infection cases have been reported, making S. parasuis a potential opportunistic zoonotic pathogen. In this study, we analysed the genomic characteristics of S. parasuis, using pan-genome analysis, and compare some phenotypic determinants such as capsular polysaccharide, integrative conjugative elements, CRISPR-Cas system and pili, and predicted the potential virulence genes by associated analysis of the clinical condition of isolated source animals and genotypes. Furthermore, to discuss the relationship with S. suis, we compared these characteristics of S. parasuis with those of S. suis. We found that the characteristics of S. parasuis are similar to those of S. suis, both of them have "open" pan-genome, their antimicrobial resistance gene profiles are similar and a srtF pilus cluster of S. suis was identified in S. parasuis genome. But S. parasuis still have its unique characteristics, two novel pilus clusters are and three different type CRISPR-Cas system were found. Therefore, this study provides novel insights into the interspecific and intraspecific genetic characteristics of S. parasuis, which can be useful for further study of this opportunistic pathogen, such as serotyping, diagnostics, vaccine development, and study of the pathogenesis mechanism.
Subject(s)
Streptococcal Infections , Streptococcus suis , Swine Diseases , Animals , Cattle , Genomics , Streptococcal Infections/veterinary , Streptococcus , Streptococcus suis/genetics , Swine , Virulence/geneticsABSTRACT
Among the numerous serotypes of Avian pathogenic Escherichia coli (APEC), O1, O2 and O78 have long been considered the predominant serogroups. O145, a pivotal serogroup in non-O157 Shiga toxin-producing Escherichia coli, has never been considered an important serogroup among APEC. The prevalence of APEC O145 was determined from the results of molecular serogrouping based on 42 sequenced isolates from Jiangsu and Guangxi Provinces in China. After realizing the potential importance of O145, 224 APEC isolates isolated from Jiangsu, Guangxi, Anhui, Shandong, Henan, Yunnan and Fujian provinces were screened using PCR amplification. The results showed that the proportion of O145 detected was 37.9 % (85/224), which was higher than those of the three traditional APEC serogroups. The virulence evaluation experiment showed that this serogroup may have stronger pathogenicity. Here, we report for the first time that O145 may be emerging as a predominant serogroup of APEC in China. The possible reasons for its prevalence and oversight were analyzed through genomic analysis. Furthermore, pangenome analysis with STEC O145 was performed to assess the potential threat to humans. The discovery of the ubiquity of O145 may not be coincidental, which may also account for the failure of vaccines that target the three major serogroups. Therefore, this newly predominant serogroup should be paid more attention and the focus should not be limited to the so-called three major APEC serogroups.
Subject(s)
Escherichia coli Infections , Escherichia coli Proteins , Shiga-Toxigenic Escherichia coli , Animals , China/epidemiology , Escherichia coli/genetics , Escherichia coli Infections/epidemiology , Escherichia coli Infections/veterinary , Escherichia coli Proteins/genetics , Serogroup , Shiga-Toxigenic Escherichia coli/geneticsABSTRACT
With the increase in antimicrobial resistance of Salmonella, phages have been paid more attention to as an alternative to antibiotics. In this study, a phage designated as SP76 was isolated from sewage. It can lyse several serotypes of Salmonella, including S. typhimurium (21/33), S. enteritidis (7/7), S. dublin (4/4), S. pullorum (2/2) and S. choleraesuis (1/2). SP76 showed a latent time of about 10 min, and maintained good lytic activity at a pH range of 3-10 and temperatures between 4 and 37 °C. Moreover, its optimal multiplicity of infection (MOI) was 0.0001. Based on the results of genomic sequence and analysis, SP76 was found to have a genome of 111,639 bp that encoded 166 predicted ORFs and belong to the Demerecviridae family, order Caudovirales. No virulence or lysogen formation gene clusters were identified in the SP76 genome. A pan-genome analysis based on 100 phages within the subfamily Markadamsvirinae indicated that SP76 had 23 core genes and 1199 accessory genes. We grouped the subfamily Markadamsvirinae and found that the main difference was in group III. In vitro bacteriostasis, experiments showed that the phage SP76 reduced planktonic bacteria by 1.52 log10 CFU/mL, and biofilms (24 h old) by 0.372 log10 CFU/mL, respectively. Thus, we isolated a safe and efficient phage that might be a good antibacterial agent.
Subject(s)
Bacteriophages , Bacteriophages/genetics , Genome, Viral , Genomics , Salmonella enteritidis , SerogroupABSTRACT
RESEARCH HIGHLIGHTSPan-RV analysis was used for the first time in the discovery of APEC-protective proteins.A total of 53 potential protective proteins were screened out.Four proteins were verified as potential vaccine candidates using western blotting.
Subject(s)
Escherichia coli Infections , Escherichia coli Vaccines , Poultry Diseases , Animals , Chickens , Escherichia coli/genetics , Escherichia coli Infections/prevention & control , Escherichia coli Infections/veterinary , Poultry Diseases/prevention & controlABSTRACT
Streptococcus suis is an important zoonotic pathogen that leads to huge economic losses in the swine industry. Because of the enormous genetic and phenotypic diversity within S. suis, it is necessary to develop effective vaccines to control this zoonotic pathogen. SBP2' is a major pili subunit in S. suis that belongs to an srtBCD pili cluster and has already been reported to be associated with the pathogenesis of this bacterium. In this study, we aimed to evaluate the immunogenicity and protective ability of SBP2'. The rSBP2' protein was expressed by an Escherichia coli expression system and emulsified with Montanide ISA 201 adjuvant to prepare the subunit vaccine. Through active immune assays, the results showed that rSBP2' exhibited good immunogenicity and could protect mice from a lethal dose challenge. Additionally, the qRT-PCR data showed that the transcription levels of cytokines associated with systemic symptoms caused by S. suis were decreased, indicating that immunization with rSBP2' could protect the host from cytokine storms caused by S. suis. Furthermore, the passive immune assay showed that the humoral immunity induced by rSBP2' played an important role against S. suis infection. Taken together, SBP2' could provide proper immune protection against S. suis challenge and could be a candidate for S. suis subunit vaccine. The results of this study could provide new ideas for the development of effective vaccines against S. suis.
Subject(s)
Fimbriae, Bacterial/immunology , Immunogenicity, Vaccine , Streptococcal Infections/prevention & control , Streptococcal Vaccines/immunology , Streptococcus suis/immunology , Animals , Escherichia coli/genetics , Mice , Mice, Inbred ICR , Serogroup , Streptococcal Infections/microbiology , Streptococcus suis/genetics , Vaccines, Synthetic/immunologyABSTRACT
Atypical enteropathogenic Escherichia coli (aEPEC) is a subgroup of EPEC, which is one of the major pathogens responsible for fatal diarrhoea in children. Compared with typical EPEC (tEPEC), aEPEC lack an EAF (EPEC adherence factor) plasmid (pEAF), which encodes a series of virulence-associated genes. The extracellular matrix (ECM) component of human cells has been reported to be an important element in the interaction between host and bacterial pathogens. In this research, a 2D-Far Western blot method was performed to identifiy the bacterial proteins that could bind to fibronectin, one of the most common constituents of ECM. A total of 17 protein spots were identified, including 4 outer membrane proteins (OMPs), namely, OmpC, OmpD, OmpX and LamB. In vitro studies were used to determine whether these OMPs were involved in the adherence process. Through indirect immunofluorescence assays, four OMPs could be observed on the surfaces of host cells. After incubating the cells with the recombinant proteins, the adhesion rate of the O55:H7 isolate was decreased. Furthermore, the deletion of OmpX and LamB can also decrease the adhesion rate of WT. Taken together, a high-throughput screening method for host ECM-binding proteins based on 2D Far-Western blot was established, and four outer membrane proteins identified by this method were found to be involved in the adherence process.
Subject(s)
Enteropathogenic Escherichia coli , Escherichia coli Infections , Escherichia coli Proteins , Blotting, Far-Western , Carrier Proteins , Child , Enteropathogenic Escherichia coli/genetics , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Fibronectins , Humans , SerogroupABSTRACT
Streptococcus suis, a major emerging pathogen in swine and humans, expresses immunoglobulin G (IgG)-binding proteins (IBPs), which contribute to the ability of organism to evasion of host immune system. The objective of this study was to identify novel pig IgG (pIgG) and human IgG (hIgG)-binding proteins and characterize the binding regions of enolase from Streptococcus suis serotype 2 (S. suis 2). Here, four pIgG-binding proteins (pIBPs) and five hIgG-binding proteins (hIBPs) were identified from S. suis 2 surface proteins by 2D-Far-western blot assays. All the newly captured proteins were expressed and further confirmed their binding activity to pIgG or hIgG by Far-western blot and dot blot. In addition to previously identified factor H, fibronectin, collagen, fibrinogen, plasminogen and laminin, we also found that both pIgG and hIgG can specifically interact with enolase. Binding assays indicated that interactions of S. suis 2 enolase with pIgG and hIgG is primarily mediated by the enolase C-terminal portion (Enolase-C, a.a. 142-432). We found that hIgG exhibited stronger binding ability to Enolase-C than pIgG. Further analysis of the C-terminal regions of enolase (Enolase-C1 and Enolase-C2) suggested that the C-terminus possessed two different binding domains with distinct host IgG proteins. Strikingly, we confirmed that pIgG interacted with the Enolase-C1 (a.a. 142-271) and hIgG interacted with the Enolase-C2 (a.a. 271-432). These observations of enolase provide interesting insights in the pathogenesis of S. suis infection.