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Heliyon Jul 2023is a rod shape, Gram-negative bacterium that causes sepsis (with a greater than 50% mortality rate), necrotizing fasciitis, gastroenteritis, skin, and soft tissue...
is a rod shape, Gram-negative bacterium that causes sepsis (with a greater than 50% mortality rate), necrotizing fasciitis, gastroenteritis, skin, and soft tissue infection, wound infection, peritonitis, meningitis, pneumonia, keratitis, and arthritis. Based on pathogenicity is categorized into three biotypes. Type 1 and type 3 cause diseases in humans while biotype 2 causes diseases in eel and fish. Due to indiscriminate use of antibiotics has developed resistance to many antibiotics so curing is dramatically a challenge. is resistant to cefazolin, streptomycin, tetracycline, aztreonam, tobramycin, cefepime, and gentamycin. Subtractive genome analysis is the most effective method for drug target identification. The method is based on the subtraction of homologous proteins from both pathogen and host. By this process set of proteins present only in the pathogen and perform essential functions in the pathogen can be identified. The entire proteome of strain ATCC 27562 was reduced step by step to a single protein predicted as the drug target. AlphaFold2 is one of the applications of deep learning algorithms in biomedicine and is correctly considered the game changer in the field of structural biology. Accuracy and speed are the major strength of AlphaFold2. In the PDB database, the crystal structure of the predicted drug target was not present, therefore the Colab notebook was used to predict the 3D structure by the AlphaFold2, and subsequently, the predicted model was validated. Potent inhibitors against the new target were predicted by virtual screening and molecular docking study. The most stable compound ZINC01318774 tightly attaches to the binding pocket of bisphosphoglycerate-independent phosphoglycerate mutase. The time-dependent molecular dynamics simulation revealed compound ZINC01318774 was superior as compared to the standard drug tetracycline in terms of stability. The availability of strain ATCC 27562 has allowed identification of drug target which will provide a base for the discovery of specific therapeutic targets against
PubMed: 37449110
DOI: 10.1016/j.heliyon.2023.e17650 -
Medicine Jul 2023The diagnostic accuracy of laboratory risk indicator for necrotizing fasciitis (LRINEC) score system in specific Vibrio vulnificus (V vulnificus ) necrotizing fasciitis...
Unreliable diagnostic accuracy of laboratory risk indicator for necrotizing fasciitis (LRINEC) score but good outcome predictor in necrotizing fasciitis due to Vibrio vulnificus : A retrospective and matched-pair study.
The diagnostic accuracy of laboratory risk indicator for necrotizing fasciitis (LRINEC) score system in specific Vibrio vulnificus (V vulnificus ) necrotizing fasciitis (NF) have not been fully investigated yet. This aim of our study is to validate the LRINEC score in patients with V vulnificus NF. A retrospective study of hospitalized patients was conducted in a hospital in southern Taiwan between January 2015 and December 2022. Clinical characteristics, variables and outcomes were compared among V vulnificus NF, non- Vibrio NF and cellulitis patients. A total of 260 patients were included, 40 in V vulnificus NF group, 80 in non- Vibrio NF group and 160 patients in cellulitis group. In V vulnificus NF group with an LRINEC cutoff score ≥ 6, the sensitivity was 35% (95% confidence interval [CI]: 29%-41%), specificity was 81% (95% CI: 76%-86%), PPV was 23% (95% CI: 17%-27%), and NPV was 90% (95% CI: 88%-92%). The AUROC for accuracy of the LRINEC score in V vulnificus NF was 0.614 (95% CI: 0.592-0.636). Multi-variable logistic regression analysis revealed that LRINEC > 8 was significantly associated with higher in-hospital mortality (adjusted odds ratio = 1.57; 95% CI: 1.43-2.08; P < .01). The LRINEC score may not be an accurate tool for V vulnificus NF. That should be used with caution as a routine diagnostic tool. However, LRINEC > 8 is significantly associated with higher mortality in V vulnificus NF patients.
Topics: Humans; Fasciitis, Necrotizing; Retrospective Studies; Cellulitis; Vibrio vulnificus; Risk Factors
PubMed: 37417615
DOI: 10.1097/MD.0000000000034207 -
PLoS Neglected Tropical Diseases Jul 2023V. vulnificus is one of the deadliest waterborne pathogens, yet little is known of the ecological and environmental forces that drive outbreaks. As a nationally...
V. vulnificus is one of the deadliest waterborne pathogens, yet little is known of the ecological and environmental forces that drive outbreaks. As a nationally notifiable disease, all cases of V. vulnificus diagnosed in the United States are reported to the state in which they occurred, as well as to the Centers for Disease Control (CDC) in Atlanta, Georgia. Given that the state of Florida is a 'hotspot' for V. vulnificus in the United States, we examined the prevalence and incidence of cases reported to the Florida Department of Health (2008-2020). Using a dataset comprised of 448 cases of disease caused by V. vulnificus infection, we identified meteorological variables that were associated with clinical cases and deaths. Combined with data from the National Oceanic and Atmospheric Administration (NOAA), we first utilized correlation analysis to examine the linear relationships between satellite meteorological measurements such as wind speed, air temperature, water temperature, and sea-level pressure. We then measured the correlation of those meteorological variables with coastal cases of V. vulnificus, including the outcome, survival, or death. We also constructed a series of logistic regression models to analyze the relationship between temporal and meteorological variables during months that V. vulnificus cases were reported versus months when V. vulnificus cases were not reported. We report that between 2008 and 2020, V. vulnificus cases generally increased over time, peaking in 2017. As water temperature and air temperature increased, so too did the likelihood that infection with V. vulnificus would lead to patient death. We also found that as mean wind speed and sea-level pressure decreased, the probability that a V. vulnificus case would be reported increased. In summary, we discuss the potential factors that may contribute to the observed correlations and speculate that meteorological variables may increase in their public health relevance in light of rising global temperatures.
Topics: Humans; Air Pressure; Temperature; United States; Vibrio Infections; Vibrio vulnificus; Wind; Tropical Climate; Weather; Florida
PubMed: 37410780
DOI: 10.1371/journal.pntd.0011461 -
Nucleic Acids Research Aug 2023Cell-free protein synthesis assays have become a valuable tool to understand transcriptional and translational processes. Here, we established a fluorescence-based...
Cell-free protein synthesis assays have become a valuable tool to understand transcriptional and translational processes. Here, we established a fluorescence-based coupled in vitro transcription-translation assay as a read-out system to simultaneously quantify mRNA and protein levels. We utilized the well-established quantification of the expression of shifted green fluorescent protein (sGFP) as a read-out of protein levels. In addition, we determined mRNA quantities using a fluorogenic Mango-(IV) RNA aptamer that becomes fluorescent upon binding to the fluorophore thiazole orange (TO). We utilized a Mango-(IV) RNA aptamer system comprising four subsequent Mango-(IV) RNA aptamer elements with improved sensitivity by building Mango arrays. The design of this reporter assay resulted in a sensitive read-out with a high signal-to-noise ratio, allowing us to monitor transcription and translation time courses in cell-free assays with continuous monitoring of fluorescence changes as well as snapshots of the reaction. Furthermore, we applied this dual read-out assay to investigate the function of thiamine-sensing riboswitches thiM and thiC from Escherichia coli and the adenine-sensing riboswitch ASW from Vibrio vulnificus and pbuE from Bacillus subtilis, which represent transcriptional and translational on- and off-riboswitches, respectively. This approach enabled a microplate-based application, a valuable addition to the toolbox for high-throughput screening of riboswitch function.
Topics: Adenine; Aptamers, Nucleotide; Fluorescence; Green Fluorescent Proteins; Nucleic Acid Conformation; Riboswitch; Cell-Free System
PubMed: 37409574
DOI: 10.1093/nar/gkad574 -
Disease Models & Mechanisms Aug 2023Transgene driven expression of Escherichia coli nitroreductase (NTR1.0) renders animal cells susceptible to the antibiotic metronidazole (MTZ). Many NTR1.0/MTZ ablation...
Transgene driven expression of Escherichia coli nitroreductase (NTR1.0) renders animal cells susceptible to the antibiotic metronidazole (MTZ). Many NTR1.0/MTZ ablation tools have been reported in zebrafish, which have significantly impacted regeneration studies. However, NTR1.0-based tools are not appropriate for modeling chronic cell loss as prolonged application of the required MTZ dose (10 mM) is deleterious to zebrafish health. We established that this dose corresponds to the median lethal dose (LD50) of MTZ in larval and adult zebrafish and that it induced intestinal pathology. NTR2.0 is a more active nitroreductase engineered from Vibrio vulnificus NfsB that requires substantially less MTZ to induce cell ablation. Here, we report on the generation of two new NTR2.0-based zebrafish lines in which acute β-cell ablation can be achieved without MTZ-associated intestinal pathology. For the first time, we were able to sustain β-cell loss and maintain elevated glucose levels (chronic hyperglycemia) in larvae and adults. Adult fish showed significant weight loss, consistent with the induction of a diabetic state, indicating that this paradigm will allow the modeling of diabetes and associated pathologies.
Topics: Animals; Zebrafish; Hyperglycemia; Metronidazole; Diabetes Mellitus; Nitroreductases; Animals, Genetically Modified
PubMed: 37401381
DOI: 10.1242/dmm.050215 -
Annals of Agricultural and... Jun 2023The aims of this study were to search for the presence of bacteria in sea snails () by using culturomics and Matrix-assisted laser desorption/ionization time-of-flight...
INTRODUCTION AND OBJECTIVE
The aims of this study were to search for the presence of bacteria in sea snails () by using culturomics and Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and the antibiotic resistance/susceptibility of the sea snails.
MATERIAL AND METHODS
The anti-microbial susceptibilities of Gram-negative bacteriawas assessed by the Kirby-Bauer disk diffusion method, the presence of the genes (mcr-1 to -5), the major carbapenemase and β-lactamase resistant genes in Gram-negative bacteria, using mPCR method and 16S rRNA sequence analysis of isolates.
RESULTS
Bacterial growth accounted for 100% and 94.2% in the samples of intestine and meat, respectively, in the snails. The main organisms identified by MALDI-TOF MS were subsp. salmonicida at 33.7%, followed by at 9.6% (10/104) and at 7.7% in meat and intestine samples. and are intrinsic or chromosomally-mediated resistant against ampicillin. No genes (-1 to -5), the major carbapenemase and β-lactamase resistant genes were found. subsp. showed very low levofloxacin and meropenem resistance levels at 2.9%. When the sequence was searched in the Blast database, the genome of isolate showed high similarity with the sequences.
CONCLUSIONS
Conclusions. The findings obtained not only provide data about the proportion of bacteria in the gut and meat of the sea snails and their antibiotic resistance/susceptibility, but also show the absence of carbapenemase, colistin, and β-lactamase resistant genes among bacterial isolates from sea snail gut microbes.
Topics: Animals; Anti-Bacterial Agents; RNA, Ribosomal, 16S; Drug Resistance, Bacterial; Anti-Infective Agents; Snails
PubMed: 37387372
DOI: 10.26444/aaem/163582 -
Frontiers in Microbiology 2023Vibriosis causes enormous economic losses of marine fish. The present study investigated the intestinal microbial response to acute infection of half-smooth tongue sole...
INTRODUCTION
Vibriosis causes enormous economic losses of marine fish. The present study investigated the intestinal microbial response to acute infection of half-smooth tongue sole with different-dose within 72 h by metagenomic sequencing.
METHODS
The inoculation amount of for the control, low-dose, moderate-dose, and high-dose groups were 0, 8.5 × 101, 8.5 × 104, and 8.5 × 107 cells/g respectively, the infected fish were farmed in an automatic seawater circulation system under a relatively stable temperature, dissolved oxygen and photoperiod, and 3 ~ 6 intestinal samples per group with high-quality DNA assay were used for metagenomics analysis.
RESULTS
The acute infections with at high, medium, and low doses caused the change of different-type leukocytes at 24 h, whereas the joint action of monocytes and neutrophils to cope with the pathogen infection only occurred in the high-dose group at 72 h. The metagenomic results suggest that a high-dose infection can significantly alter the intestinal microbiota, decrease the microbial α-diversity, and increase the bacteria from Vibrio and Shewanella, including various potential pathogens at 24 h. High-abundance species of potential pathogens such as , , , , and exhibited significant positive correlations with . The function analysis revealed that the high-dose inflection group could increase the genes closely related to pathogen infection, involved in cell motility, cell wall/ membrane/envelope biogenesis, material transport and metabolism, and the pathways of quorum sensing, biofilm formation, flagellar assembly, bacterial chemotaxis, virulence factors and antibiotic resistances mainly from Vibrios within 72 h.
DISCUSSION
It indicates that the half-smooth tongue sole is highly likely to be a secondary infection with intestinal potential pathogens, especially species from and that the disease could become even more complicated because of the accumulation and transfer of antibiotic-resistance genes in intestinal bacteria during the process of intensified infection.
PubMed: 37333647
DOI: 10.3389/fmicb.2023.1178575 -
Applied and Environmental Microbiology Jul 2023Oysters play an important role in coastal ecology and are a globally popular seafood source. However, their filter-feeding lifestyle enables coastal pathogens, toxins,...
Oysters play an important role in coastal ecology and are a globally popular seafood source. However, their filter-feeding lifestyle enables coastal pathogens, toxins, and pollutants to accumulate in their tissues, potentially endangering human health. While pathogen concentrations in coastal waters are often linked to environmental conditions and runoff events, these do not always correlate with pathogen concentrations in oysters. Additional factors related to the microbial ecology of pathogenic bacteria and their relationship with oyster hosts likely play a role in accumulation but are poorly understood. In this study, we investigated whether microbial communities in water and oysters were linked to accumulation of Vibrio parahaemolyticus, Vibrio vulnificus, or fecal indicator bacteria. Site-specific environmental conditions significantly influenced microbial communities and potential pathogen concentrations in water. Oyster microbial communities, however, exhibited less variability in microbial community diversity and accumulation of target bacteria overall and were less impacted by environmental differences between sites. Instead, changes in specific microbial taxa in oyster and water samples, particularly in oyster digestive glands, were linked to elevated levels of potential pathogens. For example, increased levels of V. parahaemolyticus were associated with higher relative abundances of cyanobacteria, which could represent an environmental vector for spp. transport, and with decreased relative abundance of and other key members of the oyster digestive gland microbiota. These findings suggest that host and microbial factors, in addition to environmental variables, may influence pathogen accumulation in oysters. Bacteria in the marine environment cause thousands of human illnesses annually. Bivalves are a popular seafood source and are important in coastal ecology, but their ability to concentrate pathogens from the water can cause human illness, threatening seafood safety and security. To predict and prevent disease, it is critical to understand what causes pathogenic bacteria to accumulate in bivalves. In this study, we examined how environmental factors and host and water microbial communities were linked to potential human pathogen accumulation in oysters. Oyster microbial communities were more stable than water communities, and both contained the highest concentrations of Vibrio parahaemolyticus at sites with warmer temperatures and lower salinities. High oyster V. parahaemolyticus concentrations corresponded with abundant cyanobacteria, a potential vector for transmission, and a decrease in potentially beneficial oyster microbes. Our study suggests that poorly understood factors, including host and water microbiota, likely play a role in pathogen distribution and pathogen transmission.
Topics: Animals; Humans; Water; Ostreidae; Vibrio parahaemolyticus; Vibrio vulnificus; Bivalvia; Bacteria
PubMed: 37318344
DOI: 10.1128/aem.00318-23 -
Microbiology Spectrum Aug 2023Enteropathogenic bacteria express two-component systems (TCSs) to sense and respond to host environments, developing resistance to host innate immune systems like...
Enteropathogenic bacteria express two-component systems (TCSs) to sense and respond to host environments, developing resistance to host innate immune systems like cationic antimicrobial peptides (CAMPs). Although an opportunistic human pathogen Vibrio vulnificus shows intrinsic resistance to the CAMP-like polymyxin B (PMB), its TCSs responsible for resistance have barely been investigated. Here, a mutant exhibiting a reduced growth rate in the presence of PMB was screened from a random transposon mutant library of V. vulnificus, and response regulator CarR of the CarRS TCS was identified as essential for its PMB resistance. Transcriptome analysis revealed that CarR strongly activates the expression of the , , and operons. In particular, the operon plays a major role in developing the CarR-mediated PMB resistance. Phosphorylation of CarR by the sensor kinase CarS is required for the regulation of its downstream genes, leading to the PMB resistance. Nevertheless, CarR directly binds to specific sequences in the upstream regions of the and operons, regardless of its phosphorylation. Notably, the CarRS TCS alters its own activation state by responding to several environmental stresses, including PMB, divalent cations, bile salts, and pH change. Furthermore, CarR modulates the resistance of V. vulnificus to bile salts and acidic pH among the stresses, as well as PMB. Altogether, this study suggests that the CarRS TCS, in responding to multiple host environmental signals, could provide V. vulnificus with the benefit of surviving within the host by enhancing its optimal fitness during infection. Enteropathogenic bacteria have evolved multiple TCSs to recognize and appropriately respond to host environments. CAMP is one of the inherent host barriers that the pathogens encounter during the course of infection. In this study, the CarRS TCS of V. vulnificus was found to develop resistance to PMB, a CAMP-like antimicrobial peptide, by directly activating the expression of the operon. Although CarR binds to the upstream regions of the and operons regardless of phosphorylation, phosphorylation of CarR is required for the regulation of the operons, resulting in the PMB resistance. Furthermore, the CarRS TCS determines the resistance of V. vulnificus to bile salts and acidic pH by differentially regulating its own activation state in response to these environmental stresses. Altogether, the CarRS TCS responds to multiple host-related signals, and thus could enhance the survival of V. vulnificus within the host, leading to successful infection.
Topics: Humans; Polymyxin B; Vibrio vulnificus; Gene Expression Profiling; Bile Acids and Salts
PubMed: 37289068
DOI: 10.1128/spectrum.00305-23