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International Journal of Microbiology 2020The aim of this study was to characterise species of water samples collected from taps, boreholes, and dams in the North West province, South Africa, and assess...
The aim of this study was to characterise species of water samples collected from taps, boreholes, and dams in the North West province, South Africa, and assess biocontrol potentials of their bacteriophages. Fifty-seven putative isolates were obtained on thiosulfate-citrate-bile-salt-sucrose agar and identified using biochemical tests and species-specific PCRs. Isolates were further characterised based on the presence of virulence factors, susceptibility to eleven antibiotics, and biofilm formation potentials. Twenty-two (38.60%) isolates were confirmed as species, comprising (45.5%, = 10), (22.7%, = 5), (13.6%, = 3), (9.1%, = 2), and (9.1%, = 2). Three of the six virulent genes screened were positively amplified; four possessed the (18.18%) and (18.18%) genes, while the gene was harboured by 3 V. (13.64%) and one (4.55%) isolate. Isolates revealed high levels of resistance to cephalothin (95.45%), ampicillin (77.27%), and streptomycin (40.91%), while lower resistances (4.55%-27.27%) were recorded for other antimicrobials. Sixteen (72.7%) isolates displayed multiple antibiotic-resistant properties. Cluster analysis of antibiotic resistance revealed a closer relationship between isolates from different sampling sites. The species displayed biofilm formation potentials at 37°C (63.6, = 14), 35°C (50%, = 11), and 25°C (36.4%, = 8). Two phages isolated in this study (vB_VpM_SA3V and vB_VcM_SA3V) were classified as belonging to the family Myoviridae based on electron microscopy. These were able to lyse multidrug-resistant and strains. These findings not only indicate the presence of antibiotic-resistant virulent species from dam, borehole, and tap water samples that could pose a health risk to humans who either come in contact with or consume water but also present these lytic phages as alternative agents that can be exploited for biological control of these pathogenic strains.
PubMed: 32831847
DOI: 10.1155/2020/8863370 -
Journal of Cellular and Molecular... Dec 2022Different growth factors can regulate stem cell differentiation. We used keratinocyte growth factor (KGF) to direct adipose-derived stem cells (ASCs) differentiation...
Different growth factors can regulate stem cell differentiation. We used keratinocyte growth factor (KGF) to direct adipose-derived stem cells (ASCs) differentiation into keratinocytes. To enhance KGF bioavailability, we targeted KGF for collagen by fusing it to collagen-binding domain from Vibrio mimicus metalloprotease (vibrioCBD-KGF). KGF and vibrioCBD-KGF were expressed in Escherichia coli and purified to homogeneity. Both proteins displayed comparable activities in stimulating proliferation of HEK-293 and MCF-7 cells. vibrioCBD-KGF demonstrated enhanced collagen-binding affinity in immunofluorescence and ELISA. KGF and vibrioCBD-KGF at different concentrations (2, 10, and 20 ng/ml) were applied for 21 days on ASCs cultured on collagen-coated plates. Keratinocyte differentiation was assessed based on morphological changes, the expression of keratinocyte markers (Keratin-10 and Involucrin), and stem cell markers (Collagen-I and Vimentin) by real-time PCR or immunofluorescence. Our results indicated that the expression of keratinocyte markers was substantially increased at all concentrations of vibrioCBD-KGF, while it was observed for KGF only at 20 ng/ml. Immunofluorescence staining approved this finding. Moreover, down-regulation of Collagen-I, an indicator of differentiation commitment, was more significant in samples treated with vibrioCBD-KGF. The present study showed that vibrioCBD-KGF is more potent in inducing the ASCs differentiation into keratinocytes compared to KGF. Our results have important implications for effective skin regeneration using collagen-based biomaterials.
Topics: Humans; Cell Differentiation; Collagen; Collagen Type I; Fibroblast Growth Factor 7; HEK293 Cells; Keratinocytes; Stem Cells
PubMed: 36412036
DOI: 10.1111/jcmm.17619 -
Journal of Fish Diseases Dec 2017During recent decades, ornamental fish have proven to be one of the fastest growing categories of pets in Europe. In this framework, we evaluated both the potential...
During recent decades, ornamental fish have proven to be one of the fastest growing categories of pets in Europe. In this framework, we evaluated both the potential pathogenic and zoonotic risks caused by 53 Vibrio cholerae non-O1/non-O139 and a Vibrio mimicus strain isolated from ornamental fish species mostly originating from South-East Asia countries between 2000 and 2015 in Italy. All the strains were firstly identified at species level by biochemical, phylogenetic and mass spectrometry (matrix-assisted laser desorption ionization time of flight) methods, and then studied to reveal the presence of the main virulence and colonization-associated factors, as ctxA, ace, zot, stn/sto, toxR, rtxA, hlyA and tcpA by multiplex and single endpoint PCR assays. Findings showed that 21 of 54 strains harboured at least one virulence factor with a predominance for the toxR , rtxA and hlyAET genotype. Interestingly, the V. mimicus strain harboured the colonization factor and the CTX prophage receptor, tcpA, indicating the ability to capture and integrate it in its genome increasing its pathogenicity. Although these enterotoxins can sporadically cause gastroenteritis, the results highlight their probable involvement in causing severe implications for public health, suggesting the need for an European microbiological monitoring.
Topics: Animals; Fish Diseases; Fishes; Italy; Vibrio cholerae non-O1; Vibrio mimicus; Virulence Factors; Zoonoses
PubMed: 28677232
DOI: 10.1111/jfd.12659 -
Antonie Van Leeuwenhoek Jun 2017The Vibrionaceae are Gram-negative bacteria present in marine and estuarine environments worldwide, including several species known as important pathogens to humans and...
The Vibrionaceae are Gram-negative bacteria present in marine and estuarine environments worldwide, including several species known as important pathogens to humans and aquatic organisms. The aim of this research was to investigate the occurrence and virulence properties of Vibrio and Salinivibrio isolated from lagoons at Cuare Wildlife Refuge and Margarita Island in the southern Caribbean Sea. Water, plankton and oyster samples were collected during October 2011 and March 2012 and examined by specific PCR and culture methods. Vibrio genus DNA was detected in 95% of samples, while the intergenic spacer region (ISR) of Vibrio cholerae and the genes that code for the thermolabile direct haemolysin (tl) of Vibrio parahaemolyticus and the haemolysin/cytolysin (vvhA) of Vibrio vulnificus were absent or amplified in low proportions (23, 5, and 0%, respectively). Nine isolates from water and plankton were confirmed as Vibrio or Salinivibrio by phenotypic tests, 16S rRNA gene sequencing and phylogenetic analysis. All the isolates presented similar patterns of virulence factors, in which the genes ctxA (encoding for cholera toxin), tl and vvhA were lacking, whereas seven isolates displayed antibiotic resistance against ampicillin and cephalosporins. The 16S rRNA phylogenetic analysis showed the clustering of Vibrio isolates in three main clades: the plankton isolate from Cuare Wildlife Refuge formed a group with V. cholerae and Vibrio mimicus while the Margarita isolates clustered with sequences from the harveyi clade and Salinivibrio. This is the first time that Salinivibrio species are reported in tropical lagoons of the Caribbean Sea with antibiotic resistance.
Topics: Animals; Caribbean Region; Humans; Phylogeny; RNA, Ribosomal, 16S; Seawater; Tropical Climate; Vibrio; Virulence; Water Microbiology
PubMed: 28324231
DOI: 10.1007/s10482-017-0856-0 -
Cureus Aug 2023Malaria is referred to as a "rainy season disease" and is brought on by species. , a spirochete, cause zoonosis leptospirosis. It is pretty uncommon for both diseases...
Malaria is referred to as a "rainy season disease" and is brought on by species. , a spirochete, cause zoonosis leptospirosis. It is pretty uncommon for both diseases to coinfect one another. Before assuming a fever is caused by a vector-borne disease, it is essential to first rule out other possible causes, regardless of the patient's risk factors. This case report demonstrates an unusual coinfection and how it manifests. The patient can avoid many deadly consequences with early detection and prompt treatment. There have been reports of coinfections between malaria and various infectious diseases, including dengue, hantavirus, and filariasis. Recently, a few case reports of coinfection with leptospirosis and malaria have also been published. Leptospirosis and malaria are both spreadable diseases that are prevalent throughout the world, particularly in the tropics. We discuss a case of coinfection with meningoencephalitis, leptospirosis, and malaria in a young male who required intensive care unit (ICU) care. It is difficult to distinguish between single infections and coinfections due to the wide variability in presentation, which may further confound the clinical features. Furthermore, when a coinfection is present but has not yet been identified, the clinical course may worsen because there is no effective treatment. This case report demonstrates the uncommon coinfection appearance and related symptoms. The case study also examined the management of patients with leptospirosis, meningoencephalitis, and life-threatening malaria coinfections as well as the clinical course of such coinfections. A meningeal infection or inflammation that resembles both meningitis and encephalitis is referred to as meningoencephalitis.
PubMed: 37746455
DOI: 10.7759/cureus.43879 -
Marine Environmental Research Jul 2021Bacteria from the Vibrio genus are a ubiquitous component of coastal and estuarine ecosystems with several pathogenic Vibrio species displaying preferences for warm...
Bacteria from the Vibrio genus are a ubiquitous component of coastal and estuarine ecosystems with several pathogenic Vibrio species displaying preferences for warm tropical waters. We studied the spatial and temporal abundance of three key human potential pathogens V. parahaemolyticus, V. cholerae and V. vulnificus in northern tropical Australia, over the wet and dry seasons, to identify environmental parameters influencing their abundance. Quantitative PCR (qPCR) analysis revealed that V. parahaemolyticus occurred more frequently and in higher abundance than V. cholerae and V. vulnificus across all locations examined. All three species were more abundant during the wet season, with V. parahaemolyticus abundance correlated to temperature and conductivity, whereas nutrient concentrations and turbidity best explained V. vulnificus abundance. In addition to these targeted qPCR analyses, we assessed the composition and dynamics of the entire Vibrio community using hsp60 amplicon sequencing. Using this approach, 42 Vibrio species were identified, including a number of other pathogenic species such as V. alginolyticus, V. mimicus and V. fluvialis. The Vibrio community was more diverse in the wet season, with temperature and dissolved oxygen as the key factors governing community composition. Seasonal differences were primarily driven by a greater abundance of V. parahaemolyticus and V. vulnificus during the wet season, while spatial differences were driven by different abundances of V. harveyi, V. campbellii, V. cholerae and V. navarrensis. When we related the abundance of Vibrio to other bacterial taxa, defined using 16S rRNA gene amplicon sequencing, V. parahaemolyticus was negatively correlated to several taxa, including members of the Rickettsiales and Saccharimonadales, while V. vulnificus was negatively correlated to Rhobacteriaceae and Cyanobiaceae. In contrast, V. alginolyticus, V. harveyi and V. mediterranei were all positively correlated to Cyanobacteria. These observations highlight the dynamic nature of Vibrio communities and expands current understanding of the processes governing the occurrence of potentially pathogenic Vibrio spp. in tropical coastal ecosystems.
Topics: Australia; Ecosystem; Humans; RNA, Ribosomal, 16S; Vibrio cholerae; Vibrio parahaemolyticus
PubMed: 34242991
DOI: 10.1016/j.marenvres.2021.105405 -
Veterinary Immunology and... Jul 2014Vibrio mimicus (V. mimicus) is an extracellular pathogen that causes ascites disease in aquatic animals. In our previous studies, the outer membrane protein U (OmpU) of...
Design and evaluation of a tandemly arranged outer membrane protein U (OmpU) multi-epitope as a potential vaccine antigen against Vibrio mimicus in grass carps (Ctenopharyngodon idella).
Vibrio mimicus (V. mimicus) is an extracellular pathogen that causes ascites disease in aquatic animals. In our previous studies, the outer membrane protein U (OmpU) of V. mimicus has been proven to be a protective antigen, and several mimotopes of the protein were identified. Here, a tandemly arranged multi-epitope peptide (named 6EPIS) was designed with six mimotopes and heterologously expressed. Then, the immunoprotection efficacy of recombinant 6EPIS (r6EPIS) was evaluated in grass carps (Ctenopharyngodon idella) by determining relative percentage survival (RPS), specific immunoglobulin M (IgM) antibody titer, and transcriptional levels of immune-related genes of inoculated grass carps. Fish vaccinated with r6EPIS via intraperitoneal injection exhibited 85.71% RPS over the control, when challenged with V. mimicus. The enzyme-linked immunosorbent assay titer of specific IgM antibodies against r6EPIS reached 1:12,800 on Day 28 post the primary immunization. After 28 days post immunization, the transcriptional level of total IgM mRNA was significantly higher in the r6EPIS-vaccinated fish than in those vaccinated with recombinant OmpU, inactivated bacterin and rHis tag peptide (p<0.05). In addition, the transcription levels of interleukin-1β and tumor necrosis factor-α genes in the spleen and head kidney of r6EPIS-vaccinated fish were significantly increased during the period of immunization and early phase of infection, while the transcription level of interleukin-10 gene was significantly increased from Day 3 to 7 post challenge, compared to the control level. These results show that r6EPIS was highly immunogenic and could elicit strong protective immune responses. It may be an attractive vaccine candidate against V. mimicus infection.
Topics: Animals; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Vaccines; Carps; Epitopes; Fish Diseases; Vibrio Infections; Vibrio mimicus
PubMed: 24751414
DOI: 10.1016/j.vetimm.2014.03.016 -
Frontiers in Microbiology 2021Water resources contaminated with pathogenic species are usually a source of devastating infection outbreaks that have been a public health concern in both developed...
Water resources contaminated with pathogenic species are usually a source of devastating infection outbreaks that have been a public health concern in both developed and developing countries over the decades. The present study assessed the prevalence of six medically significant species in some water resources in Eastern Cape Province, South Africa for 12 months. We detected vibrios in all the 194 water samples analyzed using polymerase chain reaction (PCR). The prevalence of , , , , , and in freshwater samples was 34, 19, 9, 2, 3, and 2%, and that in brackish water samples was 44, 28, 10, 7, 46, and 51%, respectively. The population of the presumptive spp. isolated from freshwater (628) and brackish water (342) samples that were confirmed by PCR was 79% (497/628) and 85% (291/342), respectively. Twenty-two percent of the PCR-confirmed isolates from freshwater ( = 497) samples and 41% of the PCR-confirmed isolates from brackish water samples ( = 291) fall among the species of interest. The incidences of , , , , , and amidst these spp. of interest that were recovered from freshwater samples were 75, 14, 4, 6, 1, and 1%, whereas those from brackish water samples were 24, 7, 3, 3, 47, and 18%, respectively. Our observation during the study suggests pollution as the reason for the unusual isolation of medically important vibrios in winter. Correlation analysis revealed that temperature drives the frequency of isolation, whereas salinity drives the composition of the targeted species at our sampling sites. The finding of the study is of public health importance going by the usefulness of the water resources investigated. Although controlling and preventing most of the factors that contribute to the prevalence of medically important bacteria, such as species, at the sampling points might be difficult, regular monitoring for creating health risk awareness will go a long way to prevent possible -related infection outbreaks at the sampling sites and their immediate environment.
PubMed: 34149632
DOI: 10.3389/fmicb.2021.617703 -
Journal of Applied Microbiology Sep 2022We aimed at developing a fast and accurate method to detect Vibrio mimicus using real-time recombinase polymerase amplification assay.
AIM
We aimed at developing a fast and accurate method to detect Vibrio mimicus using real-time recombinase polymerase amplification assay.
METHODS AND RESULTS
Specific primers and probe were designed to target V. mimicus haemolysin (vmh) gene. Target DNA was successfully amplified at 41°C within 20 min. The method exhibited a high level of specificity and the sensitivity was 2.1 × 10 copies/25 μl or 8.4 copies/μl, which is in line with real-time polymerase chain reaction (PCR). The calibration curve plotted by the second-order polynomial regression showed better than the linear curve, as the correlation coefficient was raised to 0.9907, which suggested that the second-order polynomial regressions might be considered to apply to the quantification of real-time recombinase polymerase amplification (RPA). The limit of detection (LOD) was predicted to be 77 copies/25 μl or 3 copies/μl by a probit model. The limit of quantification (LOQ) was calculated to be 28 copies /25 μl or 1 copies/μl by a receiver operating characteristic (ROC) curve, which firstly make LOQ could be available to real-time RPA. For the performance of the real-time RPA in plasma samples, the detection sensitivity of real-time RPA was as good as the real-time PCR. For pretreatment of plasma samples, the boiling method was better than using kits, as it further shortened the time of the real-time RPA in detecting V. mimicus.
CONCLUSIONS
The real-time RPA assay developed in our study shows multiple advantages over currently available DNA diagnostic method, including a quicker time-to-result for a single sample, requiring minimal infrastructure and technical support and being tolerant to inhibitors in plasma samples.
SIGNIFICANCE AND IMPACT OF THE STUDY
The real-time RPA assay developed here is a potentially valuable tool for point-of-care (POC) diagnosis of V. mimicus infection in endemic field, especially in the resources-limited settings, as combined with portable devices.
Topics: DNA Primers; Humans; Limit of Detection; Nucleic Acid Amplification Techniques; Real-Time Polymerase Chain Reaction; Recombinases; Sensitivity and Specificity
PubMed: 35702884
DOI: 10.1111/jam.15666 -
Advances in Experimental Medicine and... 2023Of the over 100 species in the genus Vibrio, approximately twelve are associated with clinical disease, such as cholera and vibriosis. Crucially, eleven of those twelve,... (Meta-Analysis)
Meta-Analysis
Of the over 100 species in the genus Vibrio, approximately twelve are associated with clinical disease, such as cholera and vibriosis. Crucially, eleven of those twelve, including Vibrio cholerae and Vibrio vulnificus, have been isolated from birds. Since 1965, pathogenic Vibrio species have been consistently isolated from aquatic and ground-foraging bird species, which has implications for public health, as well as the One Health paradigm defined as an ecology-inspired, integrative framework for the study of health and disease, inclusive of environmental, human, and animal health. In this meta-analysis, we identified 76 studies from the primary literature which report on or examine birds as hosts for pathogenic Vibrio species. We found that the burden of disease in birds was most commonly associated with V. cholerae, followed by V. metschnikovii and V. parahaemolyticus. Meta-analysis wide prevalence of our Vibrio pathogens varied from 19% for V. parahaemolyticus to 1% for V. mimicus. Wild and domestic birds were both affected, which may have implications for conservation, as well as agriculturally associated avian species. As pathogenic Vibrios become more abundant throughout the world as a result of warming estuaries and oceans, susceptible avian species should be continually monitored as potential reservoirs for these pathogens.
Topics: Animals; Humans; Vibrio; Vibrio cholerae; Vibrio vulnificus; Cholera; Birds
PubMed: 36792882
DOI: 10.1007/978-3-031-22997-8_15