-
Microorganisms Apr 2024Bacteria in the genus are ubiquitous in estuarine and coastal waters. Some species (including and are known human pathogens causing ailments like cholera, diarrhea,...
Bacteria in the genus are ubiquitous in estuarine and coastal waters. Some species (including and are known human pathogens causing ailments like cholera, diarrhea, or septicemia. Notably, can also cause a severe systemic infection (known as vibriosis) in eels raised in aquaculture facilities. Water samples were periodically collected from the estuary of the Asahi River, located in the southern part of Okayama City, Japan. These samples were directly plated onto CHROMagar Vibrio plates, and colonies displaying turquoise-blue coloration were selected. Thereafter, polymerase chain reaction was used to identify and . A total of 30 strains and 194 strains were isolated during the warm season when the water temperature (WT) was higher than 20 °C. Concurrently, an increase in coliforms was observed during this period. Notably, has two genotypes, designated as genotype 1 and genotype 2. Genotype 1 is pathogenic to humans, while genotype 2 is pathogenic to both humans and eels. The loop-mediated isothermal amplification method was developed to rapidly determine genotypes at a low cost. Of the 194 strains isolated, 80 (41.2%) were identified as genotype 1 strains. Among the 41 strains isolated when the WTs were higher than 28 °C, 25 strains (61.0%) belonged to genotype 1. In contrast, of the 32 strains isolated when the WTs were lower than 24 °C, 27 strains (84.4%) belonged to genotype 2. These results suggest that the distribution of the two genotypes was influenced by WT.
PubMed: 38792707
DOI: 10.3390/microorganisms12050877 -
Applied and Environmental Microbiology Jun 2024CRISPRi (Clustered Regularly Interspaced Palindromic Repeats interference) is a gene knockdown method that uses a deactivated Cas9 protein (dCas9) that binds a specific...
CRISPRi (Clustered Regularly Interspaced Palindromic Repeats interference) is a gene knockdown method that uses a deactivated Cas9 protein (dCas9) that binds a specific gene target locus dictated by an encoded guide RNA (sgRNA) to block transcription. Mobile-CRISPRi is a suite of modular vectors that enable CRISPRi knockdowns in diverse bacteria by integrating IPTG-inducible and genes into the genome using Tn transposition. Here, we show that the Mobile-CRISPRi system functions robustly and specifically in multiple species: , , , , and . We demonstrate efficacy by targeting both essential and non-essential genes that function to produce defined, measurable phenotypes: bioluminescence, quorum sensing, cell division, and growth arrest. We anticipate that Mobile-CRISPRi will be used in species to systematically probe gene function and essentiality in various behaviors and native environments.IMPORTANCEThe genetic manipulation of bacterial genomes is an invaluable tool in experimental microbiology. The development of CRISPRi (Clustered Regularly Interspaced Palindromic Repeats interference) tools has revolutionized genetics in many organisms, including bacteria. Here, we optimized the use of Mobile-CRISPRi in five species, each of which has significant impacts on marine environments and organisms that include squid, shrimp, shellfish, finfish, corals, and multiple of which pose direct threats to human health. The Mobile-CRISPRi technology is easily adaptable, moveable from strain to strain, and enables researchers to selectively turn off gene expression. Our experiments demonstrate Mobile-CRISPRi is effective and robust at repressing gene expression of both essential and non-essential genes in species.
Topics: Vibrio; Vibrio vulnificus; Vibrio parahaemolyticus; Gene Expression Regulation, Bacterial; CRISPR-Cas Systems; Vibrio cholerae; Clustered Regularly Interspaced Short Palindromic Repeats; Bacterial Proteins; Gene Knockdown Techniques; Aliivibrio fischeri
PubMed: 38775491
DOI: 10.1128/aem.00065-24 -
European Journal of Clinical... May 2024Non-cholera Vibrio spp. includes ubiquitous organisms living in aquatic environments. Their occurrence is associated with global warming and meteorological disasters. In...
Non-cholera Vibrio spp. includes ubiquitous organisms living in aquatic environments. Their occurrence is associated with global warming and meteorological disasters. In May 2023 the Romagna region, Italy, was affected by severe floods. In the following 15 weeks we observed 5 patients with invasive infections caused by V. vulnificus (3/5) and V. harveyi (2/5). All patients (median age 77 years) had medical comorbidities and shared exposure to seawater. Two patients needed surgery; 2 died. In conclusion, we observed an increased burden of Vibrio spp. invasive infections after May 2023 floods, affecting old patients with predisposing medical conditions.
PubMed: 38735888
DOI: 10.1007/s10096-024-04842-7 -
The Journal of Medical Investigation :... 2024Vibrio vulnificus (V. vulnificus) is a halophilic gram-negative bacterium that inhabits coastal warm water and induce severe diseases such as primary septicemia. To...
Vibrio vulnificus (V. vulnificus) is a halophilic gram-negative bacterium that inhabits coastal warm water and induce severe diseases such as primary septicemia. To investigate the mechanisms of rapid bacterial translocation on intestinal infection, we focused on outer membrane vesicles (OMVs), which are extracellular vesicles produced by Gram-negative bacteria and deliver virulence factors. However, there are very few studies on the pathogenicity or contents of V. vulnificus OMVs (Vv-OMVs). In this study, we investigated the effects of Vv-OMVs on host cells. Epithelial cells INT407 were stimulated with purified OMVs and morphological alterations and levels of lactate dehydrogenase (LDH) release were observed. In cells treated with OMVs, cell detachment without LDH release was observed, which exhibited different characteristics from cytotoxic cell detachment observed in V. vulnificus infection. Interestingly, OMVs from a Vibrio Vulnificus Hemolysin (VVH) and Multifunctional-autoprocessing repeats-in -toxin (MARTX) double-deletion mutant strain also caused cell detachment without LDH release. Our results suggested that the proteolytic function of a serine protease contained in Vv-OMVs may contribute to pathogenicity of V. vulnificus by assisting bacterial translocation. This study reveals a new pathogenic mechanism during V. vulnificus infections. J. Med. Invest. 71 : 102-112, February, 2024.
Topics: Vibrio vulnificus; Humans; Extracellular Vesicles; Hemolysin Proteins; L-Lactate Dehydrogenase; Bacterial Outer Membrane; Epithelial Cells
PubMed: 38735705
DOI: 10.2152/jmi.71.102 -
Environmental Science & Technology May 2024Climate change-induced stressors are contributing to the emergence of infectious diseases, including those caused by marine bacterial pathogens such as spp. These...
Climate change-induced stressors are contributing to the emergence of infectious diseases, including those caused by marine bacterial pathogens such as spp. These stressors alter temporal and geographical distribution, resulting in increased spread, exposure, and infection rates, thus facilitating greater -human interactions. Concurrently, wildfires are increasing in size, severity, frequency, and spread in the built environment due to climate change, resulting in the emission of contaminants of emerging concern. This study aimed to understand the potential effects of urban interface wildfire ashes on () growth and gene expression using transcriptomic approaches. was exposed to structural and vegetation ashes and analyzed to identify differentially expressed genes using the HTSeq-DESeq2 strategy. Exposure to wildfire ash altered growth and gene expression, depending on the trace metal composition of the ash. The high Fe content of the vegetation ash enhanced bacterial growth, while the high Cu, As, and Cr content of the structural ash suppressed growth. Additionally, the overall pattern of upregulated genes and pathways suggests increased virulence potential due to the selection of metal- and antibiotic-resistant strains. Therefore, mixed fire ashes transported and deposited into coastal zones may lead to the selection of environmental reservoirs of strains with enhanced antibiotic resistance profiles, increasing public health risk.
Topics: Vibrio vulnificus; Wildfires; Gene Expression
PubMed: 38690750
DOI: 10.1021/acs.est.3c08658 -
Journal of Personalized Medicine Apr 2024is a genus of Gram-negative bacteria found in various aquatic environments, including saltwater and freshwater. bacteremia can lead to sepsis, a potentially...
BACKGROUND
is a genus of Gram-negative bacteria found in various aquatic environments, including saltwater and freshwater. bacteremia can lead to sepsis, a potentially life-threatening condition in which the immune system enters overdrive in response to the disease, causing widespread inflammation and damage to tissues and organs. had the highest case fatality rate (39%) of all reported foodborne infections in the United States and a high mortality rate in Asia, including Taiwan. Numerous scoring systems have been created to estimate the mortality risk in the emergency department (ED). However, there are no specific scoring systems to predict the mortality risk of bacteremia. Therefore, this study modified the existing scoring systems to better predict the mortality risk of bacteremia.
METHODS
Cases of bacteremia were diagnosed based on the results from at least one blood culture in the ED. Patient data were extracted from the electronic clinical database, covering January 2012 to December 2021. The primary outcome was in-hospital mortality.This study used univariate and multivariate analyses to evaluate the mortality risk.
RESULTS
This study enrolled 36 patients diagnosed with bacteremia, including 23 males (63.9%) and 13 females (36.1%), with a mean age of 65.1 ± 15.7 years. The in-hospital mortality rate amounted to 25% (9/36), with 31.5% in (6/19) and 17.6% in non- (3/17). The non-survivors demonstrated higher MEDS (10.3 ± 2.4) than the survivors (6.2 ± 4.1) ( = 0.002). Concerning the qSOFA, the survivors scored 0.3 ± 0.5, and the non-survivors displayed a score of 0.6 ± 0.7 ( = 0.387). The AUC of the ROC for the MEDS and qSOFA was 0.833 and 0.599, respectively. This study modified the scoring systems with other predictive factors, including BUN and pH. The AUC of the ROC for the modified MEDS and qSOFA reached up to 0.852 and 0.802, respectively.
CONCLUSION
The MEDS could serve as reliable indicators for forecasting the mortality rate of patients grappling with bacteremia. This study modified the MEDS and qSOFA to strengthen the predictive performance of mortality risk for bacteremia. We advocate the prompt initiation of targeted therapeutic interventions and judicious antibiotic treatments to curb fatality rates.
PubMed: 38673012
DOI: 10.3390/jpm14040385 -
Asian Journal of Surgery Apr 2024
PubMed: 38658273
DOI: 10.1016/j.asjsur.2024.04.072 -
Natural Product Research Apr 2024This study was conducted to isolate and identify the bioactive compounds from the ethanolic extract of leaf against species through a bioassay-guided fractionation....
This study was conducted to isolate and identify the bioactive compounds from the ethanolic extract of leaf against species through a bioassay-guided fractionation. The ethanol extract was exposed to silica gel chromatography followed by reversed phase HPLC to isolate the most effective fraction against . Using further UHPLC-orbitrap-ion trap mass spectrometry, five compounds were isolated with broad-spectrum potency against a range of Vibrio species viz. and The IC values for the compounds ranged from 8 to 48 µg/mL against the most sensitive species and 58 to >400 µg/mL against The results of the toxicity tests demonstrated that the compounds were not harmful for shrimp. The study's findings indicate that leaf extract may contain bioactive molecules that are able to be substituted for antibiotics to treat vibriosis in shrimp farming.
PubMed: 38648539
DOI: 10.1080/14786419.2024.2344192 -
Italian Journal of Food Safety Feb 2024The majority of human diseases attributed to seafood are caused by spp., and the most commonly reported species are , , and . The conventional methods for the detection...
Detection of pathogenic spp. in foods: polymerase chain reaction-based screening strategy to rapidly detect pathogenic , , and in bivalve mollusks and preliminary results.
The majority of human diseases attributed to seafood are caused by spp., and the most commonly reported species are , , and . The conventional methods for the detection of species involve the use of selective media, which are inexpensive and simple but time-consuming. The present work aimed to develop a rapid method based on the use of multiplex real-time polymerase chain reaction (PCR) to detect , , and in bivalve mollusks. 30 aliquots of bivalve mollusks () were experimentally inoculated with two levels of , , and . ISO 21872-1:2017 was used in parallel for qualitative analysis. The limit of detection of 50% was 7.67 CFU/g for , 0.024 CFU/g for , and 1.36 CFU/g for . For and , the real-time PCR protocol was demonstrated to amplify the pathogens in samples seeded with the lowest and highest levels. The molecular method evaluated showed a concordance rate of 100% with the reference microbiological method. was never detected in samples contaminated with the lowest level, and it was detected in 14 samples (93.33%) seeded with the highest concentration. In conclusion, the developed multiplex real-time PCR proved to be reliable for and Results for are promising, but further analysis is needed. The proposed method could represent a quick monitoring tool and, if used, would allow the implementation of food safety.
PubMed: 38623280
DOI: 10.4081/ijfs.2024.11635 -
Surgical Infections May 2024infections develop rapidly and have high mortality and disability rates. can cause local wound infection, gastroenteritis, or septicemia. In this case, an 86-year-old...
infections develop rapidly and have high mortality and disability rates. can cause local wound infection, gastroenteritis, or septicemia. In this case, an 86-year-old male was accidentally stabbed in the middle of his right thumb while cleaning whitewater fish and came to the emergency department with high fever and painful swelling of the right hand. Physical examination revealed hemorrhagic bullae in the right hand. Emergency surgery and bacterial culture were performed. Because of timely antibiotic use and surgical treatment, the patient eventually recovered and was discharged from the hospital. This case suggests that the possibility of should be considered in cases of severe infection of the extremities, even without a history of seafood consumption or seawater exposure. Early recognition, rational choice of antibiotic agents, and timely wound debridement can substantially improve the prognosis of patients and reduce mortality.
Topics: Humans; Fasciitis, Necrotizing; Male; Vibrio vulnificus; Vibrio Infections; Aged, 80 and over; Sepsis; Anti-Bacterial Agents; Fingers; Debridement
PubMed: 38608243
DOI: 10.1089/sur.2023.284