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Vibrio cholerae is a cholera-causing pathogen known to instigate severe contagious diarrhea that affects millions globally. Survival of vibrios depend on a combination of multicellular responses and adapt to changes that prevail in the environment. This process is achieved through a strong communication at the cellular level, the process has been recognized as quorum sensing (QS). The severity of infection is highly dependent on the QS of vibrios in the gut milieu. The quorum may exist in a low/high cell density (LCD/HCD) state to exert a positive or negative response to control the regulatory pathogenic networks. The impact of this regulation reflects on the transition of pathogenic V. cholerae from the environment to infect humans and cause outbreaks or epidemics of cholera. In this context, the review portrays various regulatory processes and associated virulent pathways, which maneuver and control LCD and HCD states for their survival in the host. Although several treatment options are existing, promotion of therapeutics by exploiting the virulence network may potentiate ineffective antibiotics to manage cholera. In addition, this approach is also useful in resource-limited settings, where the accessibility to antibiotics or conventional therapeutic options is limited. [ABSTRACT FROM AUTHOR]

Background: Wastewater-based epidemiology (WBE) is already being adopted for the surveillance of health conditions of communities and shows great potential for the monitoring of infectious pathogens of public health importance. There is however paucity of robust data to support extensive WBE in Nigeria. This study evaluated the prevalence of clinically relevant infectious pathogens and provided antimicrobial resistance profiles of bacteria pathogens in wastewater canals in Lagos State at a single point in time. Methods: This is a cross-sectional survey of wastewater canals in 20 Local Government Areas (LGAs) in Lagos State for detection of bacteria pathogens of public health importance including non-tuberculous mycobacteria and SARS-Cov-2 virus using cultural analysis and conventional Polymerase Chain Reaction (PCR) techniques. Descriptive epidemiological survey of communities around the canals was done using questionnaires to assess exposure pathways. Statistical analysis was done using SPSS version 27 while P value of < 0.05 was considered as significant. Results: Three thousand and fifty-four (3054) questionnaires were administered to 1215 (39.8%) females and 1658 (54.3%) males in communities situated around 40 canals in 20 LGAs. Although majority (81.8%) reported using water closet toilet system and pit latrine (12.5%), a few of them admitted to open defaecation [101 (3.3%)] while 299 (9.8%) engaged in open field waste disposal. SARS-CoV-2 was not detected from wastewater in this study. Two mycobacterial species that included Mycobacterium fortitium group (13, 32.5%) and Mycobacterium kansasii (11, 27.5%) were identified in 15 out of 20 LGAs sampled. A total of 123 bacteria pathogens were isolated across the 40 canals. Prominent enteropathogens isolated included Escheriachia coli (28.5%), Salmonella spp (16.3%), Vibro cholerae (10.6%) and Shigella spp (5.7%). Extended spectrum beta-lactamase genes were prominent (87.5%) in the wastewater samples with almost a half (42.5%) of the canals containing both SHV and CTX-M. Conclusion: This study highlights the presence of pathogens with potential to cause epidemic in wastewater canals in Lagos State and provides evidence to inform policy and strategies for wastewater monitoring and treatment. Further studies involving longitudinal monitoring of time-based variations is needed to identify trends in pathogen loads and AMR patterns over time. [ABSTRACT FROM AUTHOR]

Background: The bacterium Vibrio cholerae causes diarrheal illness and can acquire genetic material leading to multiple drug resistance (MDR). Rapid detection of resistance-conferring mobile genetic elements helps avoid the prescription of ineffective antibiotics for specific strains. Colorimetric loop-mediated isothermal amplification (LAMP) assays provide a rapid and cost-effective means for detection at point-of-care since they do not require specialized equipment, require limited expertise to perform, and can take less than 30 min to perform in resource limited regions. LAMP output is a color change that can be viewed by eye, but it can be difficult to design primer sets, determine target specificity, and interpret subjective color changes. Methods: We developed an algorithm for the in silico design and evaluation of LAMP assays within the open-source PCR Signature Erosion Tool (PSET) and a computer vision application for the quantitative analysis of colorimetric outputs. First, Primer3 calculates LAMP primer sequence candidates with settings based on GC-content optimization. Next, PSET aligns the primer sequences of each assay against large sequence databases to calculate sufficient sequence similarity, coverage, and primer arrangement to the intended taxa, ultimately generating a confusion matrix. Finally, we tested assay candidates in the laboratory against synthetic constructs. Results: As an example, we generated new LAMP assays targeting drug resistance in V. cholerae and evaluated existing ones from the literature based on in silico target specificity and in vitro testing. Improvements in the design and testing of LAMP assays, with heightened target specificity and a simple analysis platform, increase utility for in-field applications. Overall, 9 of the 16 tested LAMP assays had positive signal through visual and computer vision-based detection methods developed here. Here we show LAMP assays tested on synthetic AMR gene targets for aph(6), varG, floR, qnrVC5, and almG, which allow for resistance to aminoglycosides, penicillins, carbapenems, phenicols, fluoroquinolones, and polymyxins respectively. [ABSTRACT FROM AUTHOR]

Non-O1/non-O139 Vibrio cholerae (NOVC) strains are a distinct group of Vibrio cholerae that do not cause epidemic cholera. NOVC infections usually cause mild forms of gastroenteritis, and rarely severe (extra)intestinal infections, mostly affecting immunocompromised patients. Here, we describe the clinical course of a patient with NOVC bacteremia causing multiple liver abscesses, after drinking from a freshwater well in a non-coastal area. This case highlights the potential of a V.cholerae strain, that is phylogenetically distinct from the current pandemic cholera strain, to cause severe extra-intestinal infections, including liver abscesses. [ABSTRACT FROM AUTHOR]

Ballast water is essential for ship operations, but can also transport harmful organisms between ports, threatening local environments. The Ballast Water Management (BWM) Convention, established by the International Maritime Organization (IMO), requires ships to implement ballast water management measures to address this issue. In this study, ballast water samples were collected from ships entering Shahid Rajaee Port in Iran before and after the Ballast Water Management Convention came into force in 2017. The sampling was conducted in coordination with Iranian authorities, following IMO guidelines. Vibrio cholerae, Escherichia coli, and Enterococci were identified in 97 ships, and physical parameters such as temperature, salinity, and pH were measured in 15 ships. To prevent sample contamination, rigorous protocols were followed, including using sterile equipment, appropriate storage, and immediate transfer to the lab. The results showed that before the BWM Convention, V. cholerae was found in 6 out of 14 ships (42%), with ballast water retention times of 2–58 days. After the convention, V. cholerae was found in only 2 out of 83 ships (2.4%), with ballast water retention of 2–3 days. This indicates a significant reduction in the risk of pathogen transfer through ballast water. Further analysis after the Convention showed that V. cholerae was initially detected in 4 out of 15 ships, but only 1 ship (6.6%) had levels above the standard. E. coli and Enterococci were also detected in multiple ships, but their levels were below the standard. The study investigated the relationship between the bacterial levels and the physical parameters. While some correlations were found between E. coli/Enterococci and parameters like pH, temperature, salinity, and water retention time, no significant impact of physical parameters on V. cholerae levels was observed. T-tests revealed significant relationships between the bacterial levels and the physical parameters, as well as the ballast water retention time. Ships were found to have followed the ballast water exchange guidelines of exchanging at least 200 or 50 nautical miles from shore and at depths over 200 m, as mapped out for the Persian Gulf region. The results indicate that the implementation of the BWM Convention has been effective in reducing the risk of pathogen transfer through ballast water. However, the study emphasizes the importance of ongoing monitoring and enforcement to ensure continued compliance with the convention's requirements. [ABSTRACT FROM AUTHOR]

Abstract Non-O1/non-O139 Vibrio cholerae (NOVC) strains are a distinct group of Vibrio cholerae that do not cause epidemic cholera. NOVC infections usually cause mild forms of gastroenteritis, and rarely severe (extra)intestinal infections, mostly affecting immunocompromised patients. Here, we describe the clinical course of a patient with NOVC bacteremia causing multiple liver abscesses, after drinking from a freshwater well in a non-coastal area. This case highlights the potential of a V. cholerae strain, that is phylogenetically distinct from the current pandemic cholera strain, to cause severe extra-intestinal infections, including liver abscesses.

Copyright of Journal of Food Safety & Quality is the property of Journal of Food Safety & Quality Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Vibrio cholera (V.C.) is currently one of the most hazardous bacteria since it can lead to hepatitis, diarrhea, vomiting, and typhoid. The various methodologies include time-consuming steps that take several hours before the test report is accessible. Therefore, creating a quick and reliable technique to identify V.C. is essential. The detection of V.C. is proposed in this study using a unique bio-photonic detector that employs Surface Plasmon Resonance (SPR) structure and layers of silver (Ag), silicon (Si), and graphene (G). The reflectance spectrum properties are inspected by the transfer matrix technique. To optimize the efficiency of the bio-photonic sensor, the effects of changing the type of prism and layer thicknesses are thoroughly examined. The proposed detector achieves an extremely great sensitivity of 307.81 Deg./RIU as the structure has Ag (50 nm), Si (3 nm), and the G monolayer, which is not obtained in many previous studies in this sector. The suggested detector also facilitates designing for industry using low-cost product nanofabrication methods due to its low cost, customizable design and straightforward structure. Based on these benefits, the suggested detector candidate is a practical and effective device for detecting V.C. in water, and it may also be used for any biological sample. The optical device designed based on the proposed sensor can be used for detecting V.C and used in biomedical applications. [ABSTRACT FROM AUTHOR]

Purpose: Waterborne pathogens pose a significant threat to public health, emphasizing the continuous necessity for advancing robust detection techniques, particularly in preventing outbreaks associated with these pathogens. This study focuses on cholera, an infectious disease caused by Vibrio cholerae, serogroups O1 and O139, often transmitted through contaminated water and food, raising significant public health concerns in areas with poor sanitation and limited access to clean water. Methods: We developed a colorimetric biosensor using aptamer-functionalized gold nanoparticles to identify Vibrio cholerae O139 and address this issue. The detection mechanism relies on the color change of gold nanoparticles (AuNPs) from red to blue-purple induced by NaCl after the pathogen incubation and aptamer-target binding. Initial steps involved synthesizing and characterizing AuNPs, then exploring the impact of aptamer and NaCl concentrations on AuNP agglomeration. Optimization procedures for aptamer concentration and salt addition identified the optimal conditions for detection as 120 pM aptamers and 1 M NaCl. Results: The aptasensor demonstrated a robust linear relationship, detecting V. cholerae concentrations from 103 to 108 CFU/mL, with a limit of detection (LOD) of 587 CFU/mL. Specificity tests and accurate sample analyses confirmed the efficiency of the AuNPs aptasensor, showcasing its reliability and speed compared to traditional culture examination methods. Moreover, we extended the aptasensor to a paper-based sensing platform with similar detection principles. Conclusion: The change in color upon target binding was captured with a smartphone and analyzed using image processing software. The paper-based device detected the target in less than 2 min, demonstrating its convenience for on-field applications. [ABSTRACT FROM AUTHOR]

Introduction. The evolutionary association of Vibrio cholerae with chitin provided resistance to stress and protection from predators. The most important mechanism that provided V. cholerae with the effectiveness of association with chitin is biofilm formation. The ability to form a biofilm in V. cholerae depends on the presence of a factor, toxin-corrected adhesion pili (TCP), which are synthesized by the tcp A-F genes. One of the key methods for studying biofilms is microscopy. It allows one to visualize the structural elements and study various parameters of biofilms and the effects of various factors on them. Aim. To determine the epidemiological significance of the biofilm-forming ability of toxigenic strains by their morphological characteristics on chitin-containing substrates. Study of structural differences in biofilms of Vibrio cholerae tcpA+– and tcpA– strains on chitin-containing substrates. Results. It has been shown that Vibrio cholerae tcpA+– and tcpA– strains are able to form biofilms on the surface of chitin-containing substrates. The intensity of biofilm formation is more pronounced in tcpA+ strains, because V. cholerae ctxA+ tcpA+ cells in the biofilm are predominantly singly located and the surface of the chitinous exoskeleton with which they are in contact is intact, V. cholerae ctxA– tcpA– cells form chains in the biofilm, which indicates division processes, and scattered chitin of the endocuticle indicates activity of metabolic processes. Conclusion. The strains of V. cholerae used in the work, regardless of the presence or absence of the ctx and tcp genes, form bioplecs on a chitin substrate. The indicator of biofilm formation in terms of the thickness of the biofilm matrix is higher in V. cholerae ctxA+ tcpA+ , in terms of the degree of degradation of the chitin substrate it is higher in V. cholerae ctxA– tcpA– .

This paper present graphene based metasurface circular resonator (MSCR) inspired optical sensors for detecting waterborne bacteria such as E. coli, Vibrio Cholera, and Shigella flexneri. The optimal design is achieved through simulating all possible combination of proposed structure and analysis based on evaluation metrices. In this study, we have investigated four different sensors and among these sensors other variations are also examined by varying the graphene chemical potential to finalize the structure. The sensor design also takes into account cost-effectiveness, ease of use, and scalability to ensure the widespread adoption of the technology. The proposed designs consist of one central circle surrounded by multiple circles that increase in number with each design. These designs provide a larger surface area for bacterial adhesion, leading to increased detection sensitivity. The MSCR designs can be easily fabricated using standard techniques and can be modified to detect other types of bacteria or contaminants by changing the surface chemistry. [ABSTRACT FROM AUTHOR]

To ensure good water quality, microbiological contamination in water must be detected. This process is made easier and more distinctive using a photonic crystal fiber (PCF), which offers outstanding optical sensing capabilities. Herein, a PCF sensor model is proposed for detecting two types of waterborne bacteria, namely, Vibrio cholera and Escherichia coli bacteria. The core region of the proposed PCF sensor is made up of a single rectangle and the cladding region has 32 rectangular air holes that have the same height and width as the core rectangle. Zeonex is employed as the fiber material. Using Comsol 5.6 which is based on the finite‐element method, the model is numerically analyzed and structured. The simulation verifies the effectiveness of the proposed PCF to detect the analyte samples. Numerous performance indicators are calculated at an operating 2.8 THz. Simulation results show that the proposed PCF sensor is promising. Extremely high relative sensitivity (97.996%), lower effective area (6.3575 × 104 μm2), higher numerical aperture (0.23319), lower effective material loss (0.0034 cm−1), and lower confinement loss (0.1 × 10−14) are obtained which indicate an efficient PCF sensor. Additionally, the simplicity of the PCF design ensures the fabrication possibilities of the proposed sensor. [ABSTRACT FROM AUTHOR]

Genomic information can be used to predict major pathogenic traits of pathogens without the need for laboratory experimentation. However, no Vibrio cholerae genome-based trait identification tools currently exist. The aim of this study was to develop a web-based prediction tool to identify Vibrio pathogenic traits using publicly available 796 whole-genome sequences of V. cholerae. Using this application, 68 structural O-antigen gene clusters belonging to 49 serogroups of V. cholerae were classified, and the composition of the genes within the O-antigen cluster of each serogroup was identified. The arrangement and location of the CTX prophage and related elements of the seventh cholera pandemic strains were also revealed. With the versatile tool, named VicPred, we analyzed the assemblage of various SXTs (sulfamethoxazole/trimethoprim resistance element) and major genomic islands (GIs) of V. cholerae, and the increasing trend in drug-resistance revealing high resistance of the V. cholerae strains to certain antibiotics. The pathogenic traits of newly sequenced V. cholerae strains could be analyzed based on these characteristics. The accumulation of further genome data will expedite the establishment of a more precise genome-based pathogenic traits analysis tool.

This study aimed to investigate whether the VCA0560 gene acts as an active diguanylate cyclase (DGC) in Vibrio cholerae and how its transcription is regulated by Fur and HapR. The roles of VCA0560 was investigated by utilizing various phenotypic assays, including colony morphological characterization, crystal violet staining, Cyclic di-GMP (c-di-GMP) quantification, and swimming motility assay. The regulation of the VCA0560 gene by Fur and HapR was analyzed by luminescence assay, electrophoretic mobility shift assay, and DNase I footprinting. VCA0560 gene mutation did not affect biofilm formation, motility, and c-di-GMP synthesis in V. cholerae , and its overexpression remarkably enhanced biofilm formation and intracellular c-di-GMP level but reduced motility capacity. The transcription of the VCA0560 gene was directly repressed by Fur and the master quorum sensing regulator HapR. Overexpressed VCA0560 functions as an active DGC in V. cholerae , and its transcription is repressed by Fur and HapR. [ABSTRACT FROM AUTHOR]

The two‐component regulatory system (TCS) is a major regulatory system in bacteria that occurs in response to environmental changes and involves the sensor histidine kinase (HK) protein and response regulator (RR) protein. Among the TCSs, PhoR/PhoB is crucial for bacteria to adapt to changes in environmental phosphate concentrations. In addition, recent studies have shown that PhoR binding to the MgtC virulence factor activates phosphate transport for normal pathogenesis. In this work, we determined the crystal structure of the catalytic ATP binding domain of the PhoR sensor histidine kinase from Vibrio cholera, compared the structure with the known HK protein structures and discussed the potential binding interface with MgtC. [ABSTRACT FROM AUTHOR]

Bacteriophage therapy targeting the increasingly resistant Vibrio cholerae is highly needed. Hence, studying the phenotypic behavior of potential phages under different conditions is a prerequisite to delivering the phage in an active infective form. The objective of this study was to characterize phage VP4 (vB_vcM_Kuja), an environmental vibriophage isolated from River Kuja in Migori County, Kenya in 2015. The phenotypic characteristics of the phage were determined using a one‐step growth curve, restriction digestion profile, pH, and temperature stability tests. The results revealed that the phage is stable through a wide range of temperatures (20−50°C) and maintains its plaque‐forming ability at pH ranging from 6 to 12. The one‐step growth curve showed a latent period falling between 40 and 60 min, while burst size ranged from 23 to 30 plaque‐forming units/10 µl at the same host strain. The restriction digestion pattern using EcoRI, SalI, HindIII, and XhoI enzymes showed that HindIII could cut the phage genome. The phage DNA could not be restricted by the other three enzymes. The findings of this study can be used in future studies to determine phage−host interactions. [ABSTRACT FROM AUTHOR]

Objective: Cholera is a challenging ancient disease caused by Vibrio cholera (V. cholera). Antibiotics that prevent cell wall synthesis are among the first known antibiotic groups. Due to its high consumption, V. cholera has developed resistance to the majority of antibiotics in this class. Resistance to recommended antibiotics for the treatment of V. cholera has also increased. In light of the decrease in consumption of certain antibiotics in this group that inhibit cell wall synthesis and the implementation of new antibiotics, it is necessary to determine the antibiotic resistance pattern of V. cholera and to employ the most effective treatment antibiotic. Method: An comprehensive systematic search for relevant articles was conducted in PubMed, Web of Science, Scopus, and EMBASE through October 2020. Stata version 17.1 utilized the Metaprop package to execute a Freeman-Tukey double arcsine transformation in order to estimate weighted pooled proportions. Results: A total of 131 articles were included in the meta-analysis. Ampicillin was the most investigated antibiotic. The prevalence of antibiotic resistance was in order aztreonam (0%), cefepime (0%), imipenem (0%), meropenem (3%), fosfomycin (4%), ceftazidime (5%), cephalothin (7%), augmentin (8%), cefalexin (8%), ceftriaxone (9%), cefuroxime (9%), cefotaxime (15%), cefixime (37%), amoxicillin (42%), penicillin (44%), ampicillin (48%), cefoxitin (50%), cefamandole (56%), polymyxin-B (77%), carbenicillin (95%) respectively. Discussion: Aztreonam, cefepime, and imipenem are the most efficient V. cholera cell wall synthesis inhibitors. There has been an increase in resistance to antibiotics such as cephalothin, ceftriaxone, amoxicillin, and meropenem. Over the years, resistance to penicillin, ceftazidime, and cefotaxime, has decreased. [ABSTRACT FROM AUTHOR]

Objective: Cholera is a challenging ancient disease caused by Vibrio cholera (V. cholera). Antibiotics that prevent cell wall synthesis are among the first known antibiotic groups. Due to its high consumption, V. cholera has developed resistance to the majority of antibiotics in this class. Resistance to recommended antibiotics for the treatment of V. cholera has also increased. In light of the decrease in consumption of certain antibiotics in this group that inhibit cell wall synthesis and the implementation of new antibiotics, it is necessary to determine the antibiotic resistance pattern of V. cholera and to employ the most effective treatment antibiotic.Method: An comprehensive systematic search for relevant articles was conducted in PubMed, Web of Science, Scopus, and EMBASE through October 2020. Stata version 17.1 utilized the Metaprop package to execute a Freeman-Tukey double arcsine transformation in order to estimate weighted pooled proportions.Results: A total of 131 articles were included in the meta-analysis. Ampicillin was the most investigated antibiotic. The prevalence of antibiotic resistance was in order aztreonam (0%), cefepime (0%), imipenem (0%), meropenem (3%), fosfomycin (4%), ceftazidime (5%), cephalothin (7%), augmentin (8%), cefalexin (8%), ceftriaxone (9%), cefuroxime (9%), cefotaxime (15%), cefixime (37%), amoxicillin (42%), penicillin (44%), ampicillin (48%), cefoxitin (50%), cefamandole (56%), polymyxin-B (77%), carbenicillin (95%) respectively.Discussion: Aztreonam, cefepime, and imipenem are the most efficient V. cholera cell wall synthesis inhibitors. There has been an increase in resistance to antibiotics such as cephalothin, ceftriaxone, amoxicillin, and meropenem. Over the years, resistance to penicillin, ceftazidime, and cefotaxime, has decreased.

Copyright of Public Health Forum is the property of De Gruyter and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Last cholera epidemic has been recorded in Bangladesh between 1992–1993, while few sporadic localized outbreaks have been reported as recent as 2005. Serotype O1 of Vibrio cholera is considered as the principal causative agent which transmits through contaminated drinking water resulting that epidemic. Therefore, the objective of this research was to isolate V. cholera in 3 different water sources; River, pond and tube-well, in 5 different locations of Gazipur, Bangladesh, and to analyze their antibiogram study. A total of 45 water samples were randomly collected for the isolation and identification of Vibrio spp. Samples are then serially diluted in alkaline peptone water and streak on Thiosulfate Citrate Bile Salt Sucrose-TCBS agar for quantification of V. spp. For V. cholera isolation water samples were first enriched in nutrient broth at 37 °C for 16 hours followed by cultivation in selective media; TCBS agar at 37 °C for 24 hours. Yellow colonies on TCBS agar were screed as V. cholera and was confirmed by analyzing their biochemical characteristics like Catalase, Oxidase, MR, VP, Indole, Sugar fermentation. Following isolation antibiotic sensitivity test was performed on each V. cholera isolates to determine their antibiotic sensitivity profile. The results showed, out of 45 samples 12 contained V. cholera. Tube-well water has significantly lower concentration (log CFU/mL) of V. spp. than river and pond water (P < 0.05). Bacterial concentration doesn't deviate (P > 0.05) significantly in 5 different location the sample was collected from. All the 12 isolates were sensitive to Gentamicin and ciprofloxacin (100%), while Chloramphenicol (91.67%), Sulfamethoxazole (91.67%), Azithromycin (66.67%) showed high sensitivity. Isolates showed marginal sensitivity towards Tetracycline (33.33%), and Cephalexin (16.67%) and 100% resistance against antibiotics like Vancomycin, Penicillin, Erythromycin, and Nalidixic Acid. Based on these data we recommend using tube-well water instead of river and pond water for drinking purposes. Furthermore, we suggest selective use of sensitive antimicrobials listed here for therapeutics of cholera outbreak.

Background: Nanotechnology is considered one of the best advanced technologies in treatment through its ability to transport effective drugs and compounds and release them more effectively. MgO nanoparticles showed antimicrobial properties against both Gram-negative and Grampositive bacteria in vitro, including Escherichia coli and Staphylococcus aureus, as Vibrio cholera bacteria was selected during the period when the epidemic began to appear in Syria, with the presence of several people showing symptoms of diarrhea in the city of Aleppo. Methods: In this research, Nano magnesium oxide particles were prepared by sol-gel method, then the active compounds were extracted from the Syrian Inula plant using ethanol as a solvent. next step was load the plant extract on the surface of the prepared magnesium oxide particles. The antibacterial activity of the new product was studied against the bacteria that cause cholera. Results: Its effectiveness was compared with the effectiveness of 7 different antibiotics, and it outperformed all of them, noting that the studied isolate was resistant to three of them. Conclusion: The combination of nanoparticles with the natural extract is more effective in fighting bacteria, because each of them works in a different way and affects differently, so the combination of the two technologies gives greater effectiveness. [ABSTRACT FROM AUTHOR]

Vibrio cholerae includes strains responsible for the cholera disease and is a natural inhabitant of aquatic environments. V. cholerae possesses a unique polar flagellum essential for motility, adhesion, and biofilm formation. In a previous study, we showed that motility and biofilm formation are altered in the presence of subinhibitory concentrations of polymyxin B in V. cholerae O1 and O139. In this study, we performed an experimental evolution to identify the genes restoring the motility in the presence of a subinhibitory concentration of polymyxin B. Mutations in five genes have been identified in three variants derived from two different parental strains A1552 and MO10: ihfA that encodes a subunit of the integration host factor (IHF), vacJ (mlaA) and mlaF, two genes belonging to the maintenance of the lipid asymmetry (Mla) pathway, dacB that encodes a penicillin-binding protein (PBP4) and involved in cell wall synthesis, and ccmH that encodes a c-type cytochrome maturation protein. We further demonstrated that the variants derived from MO10 containing mutations in vacJ, mlaF, and dacB secrete more and larger membrane vesicles that titer the polymyxin B, which increases the bacterial survival and is expected to limit its impact on the bacterial envelope and participate in the flagellum's retention and motility. [ABSTRACT FROM AUTHOR]

Objectives: Few studies have evaluated determinants of multidrug-resistant (MDR) Vibrio cholerae O1 in older children and adults. This study aimed to characterize the prevalence of MDR V. cholerae O1 and associated risk factors among patients over five years of age in Bangladesh. Methods: Stool culture and antimicrobial susceptibility testing were performed as a part of a larger study at Dhaka Hospital in Bangladesh from March 2019–March 2020. Univariate statistics and multiple logistic regression were used to assess the association between a range of variables and MDR V. cholerae O1. Results: MDR was found in 175 of 623 (28.1%) V. cholerae O1 isolates. High levels of resistance were found to erythromycin (99.2%), trimethoprim-sulfamethoxazole (99.7%), and ampicillin (88.9%), while susceptibility was high to tetracyclines (99.7%), azithromycin (99.2%), ciprofloxacin (99.8%), and cephalosporins (98.6%). MDR was associated with prior antibiotic use, longer transport time to hospital, higher income, non-flush toilet use, greater stool frequency, lower blood pressure, lower mid-upper arm circumference, and lower percent dehydration. Conclusions: MDR V. cholerae O1 was common among patients over five in an urban hospital in Bangladesh. Significant factors associated with MDR may be actionable in identifying patients with a high likelihood of MDR.

Vibrio cholerae includes strains responsible for the cholera disease and is a natural inhabitant of aquatic environments. V. cholerae possesses a unique polar flagellum essential for motility, adhesion, and biofilm formation. In a previous study, we showed that motility and biofilm formation are altered in the presence of subinhibitory concentrations of polymyxin B in V. cholerae O1 and O139. In this study, we performed an experimental evolution to identify the genes restoring the motility in the presence of a subinhibitory concentration of polymyxin B. Mutations in five genes have been identified in three variants derived from two different parental strains A1552 and MO10: ihfA that encodes a subunit of the integration host factor (IHF), vacJ (mlaA) and mlaF, two genes belonging to the maintenance of the lipid asymmetry (Mla) pathway, dacB that encodes a penicillin-binding protein (PBP4) and involved in cell wall synthesis, and ccmH that encodes a c-type cytochrome maturation protein. We further demonstrated that the variants derived from MO10 containing mutations in vacJ, mlaF, and dacB secrete more and larger membrane vesicles that titer the polymyxin B, which increases the bacterial survival and is expected to limit its impact on the bacterial envelope and participate in the flagellum’s retention and motility.

Broad-spectrum amino acid racemases (Bsrs) enable bacteria to generate non-canonical D-amino acids (NCDAAs), whose roles and impact on microbial physiology, including modulation of cell wall structure and dissolution of biofilms, are just beginning to be appreciated. Here we used a diverse array of structural, biochemical and molecular simulation studies to define and characterize how BsrV is post-translationally regulated. We discovered that contrary to Vibrio cholerae alanine racemase AlrV highly compacted active site, BsrV’s is broader and can be occupied by cell wall stem peptides. We found that peptidoglycan peptides modified with NCDAAs are better stabilized by BsrV’s catalytic cavity and show better inhibitory capacity than canonical muropeptides. Notably, BsrV binding and inhibition can be recapitulated by undigested peptidoglycan sacculi as it exists in the cell. Docking simulations of BsrV binding the peptidoglycan polymer generate a model where the peptide stems are perfectly accommodated and stabilized within each of the dimeŕs active sites. Taking these biochemical and structural data together, we propose that inhibition of BsrV by peptidoglycan peptides underlies a negative regulatory mechanism to avoid excessive NCDAA production. Our results collectively open the door to use “à la carte” synthetic peptides as a tool to modulate DAAs production of Bsr enzymes.

In September 2019, after a reported death due to acute diarrheal disease in Shahpur village, Panchkula district, Haryana state, India, we conducted an outbreak investigation to identify the etiological agent, estimate the burden of disease, and make recommendations to prevent future outbreaks. The suspected cholera case was a resident of Shahpur huts, ≥1 year of age having ≥3 loose stools within a 24-h period between September 1 and 28, 2019 and a laboratory-confirmed cholera case, whose stool specimen tested positive for Vibrio cholerae. We identified 196 suspected cholera cases with a median age of 18 years (range: 1–65 years); 54% (106) being female. The overall attack rate was 8% (196/2,602), and the case fatality rate was 1% (2/196). Tested samples of water from tanks (n = 6), sewage effluent (n = 2), and 22% (4/18) of stool specimens collected from suspected cases were positive for V. cholerae. Strengthening surveillance, improving water, and sanitation systems are recommended to prevent future cholera outbreaks.

Water a vital resource of life has been faced with concerns of usage safety. As a strategy toward alleviating user safe-water-for-all (SWA) concern amongst safe water deprived populations, a cross-sectional analysis of various water sources were determined. These include sampling of wastewater treatment plants final effluents (WWTPFE), receiving water shed and domestic water, determination of physicochemical parameters (PPA), vibrio-colonial density (VCD), virulent/antibiotic resistant gene (V/ARG) detection and genotypic characterisation of Vibrio cholerae. Various PPA were reported to be higher above expected regulatory standard in the receiving water shed. The presumptive Vibrio counts of samples ranged between 1.10 - 7.91 log10 CFU/ml. PCR confirmed V. cholerae isolates were 61/8.04% in addition to diverse V/ARGs in various sites. The mean VCD ranges from 0.00±0.00 to 244.61±44.86 CFU/100 ml with lethality characteristic index (LCI) revealing high contamination, while detected virulent/resistant dynamics spans across gastro-enteric/sporadic, extended spectrum betalactamase (ESβL), New Delhi metalobatalactamase (NDM-β-1) and epidemic genes. Statistical significance was observed in VCD and PPA of sampled water plants at p

Bacteria have evolved diverse strategies to ensure survival under nutrient-limited conditions, where rapid energy generation is not achievable. Here, we performed a transposon insertion site sequencing loss-of-function screen to identify Vibrio cholerae genes that promote the pathogen's fitness in stationary phase. We discovered that the Mla (maintenance of lipid asymmetry) pathway, which is crucial for transferring phospholipids from the outer to the inner membrane, is critical for stationary phase fitness. Competition experiments with barcoded and fluorophore labeled wild-type and mlaE mutant V. cholerae revealed that the Mla pathway promotes re-expansion from 48h stationary phase cultures. The mutant's defect in transitioning out of stationary phase into active growth (culturability) was also observed in monocultures at 48h. However, by 96h the culturability of the mutant and wild-type strains were equivalent. By monitoring the abundances of genomically barcoded libraries of wild-type and ∆ mlaE strains, we observed that a few barcodes dominated the mutant culture at 96h, suggesting that the similarity of the population sizes at this time was caused by expansion of a subpopulation containing a mutation that suppressed the mlaE mutant's defect. Whole genome sequencing revealed that mlaE suppressors inactivated flagellar biosynthesis. Additional mechanistic studies support the idea that the Mla pathway is critical for the maintenance of V. cholerae's culturability as it promotes energy homeostasis, likely due to its role in regulating outer membrane vesicle shedding. Together our findings provide insights into the cellular processes that control re-expansion from stationary phase and demonstrate a previously undiscovered role for the Mla pathway.

Vibrio Cholera, a gram negative bacterium, is notoriously known to cause diarrheal epidemics. The serotypes O1 and O139 are mainly responsible for the diarrheal outbreaks due to the enterotoxin they produce. This enterotoxin however seems to be protective against bacteremia and hence bacteremia is rarely encountered. We report a case of an immune-competent young female who initially presented with hypokalemic periodic paralysis secondary to acute on chronic diarrhea. She was discharged after rehydration and bounced back in septic shock and her blood cultures grew Vibrio Cholera identified on Vitek. She succumbed to this organism within 24 hours of re-admission., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

Genomic information can be used to predict major pathogenic traits of pathogens without the need for laboratory experimentation. However, no Vibrio cholerae genome-based trait identification tools currently exist. The aim of this study was to develop a web-based prediction tool to identify Vibrio pathogenic traits using publicly available 796 whole-genome sequences of V. cholerae. Using this application, 68 structural O-antigen gene clusters belonging to 49 serogroups of V. cholerae were classified, and the composition of the genes within the O-antigen cluster of each serogroup was identified. The arrangement and location of the CTX prophage and related elements of the seventh cholera pandemic strains were also revealed. With the versatile tool, named VicPred, we analyzed the assemblage of various SXTs (sulfamethoxazole/trimethoprim resistance element) and major genomic islands (GIs) of V. cholerae, and the increasing trend in drug-resistance revealing high resistance of the V. cholerae strains to certain antibiotics. The pathogenic traits of newly sequenced V. cholerae strains could be analyzed based on these characteristics. The accumulation of further genome data will expedite the establishment of a more precise genome-based pathogenic traits analysis tool.

Background: Cholera is a primordial disease caused by Vibrio cholerae which existed from centuries in different parts of the world and still shows its periodic, endemic and epidemic presence. Thousands of cholera cases are reported from different parts of India and the disease remains endemic throughout the year. At present, we do not have enough knowledge about the phenotypic nature of the circulating V. cholerae strains in this part of the world. Objectives: This study was carried out over a period of 6 years with the aim defer with the changes in the prevalence and distribution of biotypes, serotypes and phage types of V. cholerae clinical isolates from various endemic regions of the country to determine phenotypic characteristics of the circulating strains and also to predict the attributes of cholera strains responsible for causing significant outbreaks in future. Materials and Methods: A total of 1882 V.cholerae O1 isolates from different cholera endemic areas of India were included in this study. V.cholerae strains which were identified as O1 biotype ElTor further analyzed for serotype and phage types using the standard methodologies. Polyvalent O1 and monospecific Inaba and Ogawa antisera were used for serotyping. A panel of five phages of Basu and Mukherjee phage typing scheme and five phages from the new phage typing scheme were used for phage typing analysis following standard methodology. Results: Maximum numbers of strains were isolated from cholera-endemic states like Gujarat and Maharashtra. All the isolates were confirmed as V. cholerae O1 biotype ElTor and majority of them were serotype Ogawa (93.2%). New phage typing scheme resulted in almost 100% typeable V. cholerae O1 strains included in this study and phage type 27 was the predominant type. Although 80% of the strains used in this study were sensitive to all the vibrio phages, S5 phage was found most efficient in lysing cholera strains indicating its broader host range. Conclusion: The current study identified phage type 27 as the most dominant type and serotype Ogawa was found continuous in circulation throughout the year which has caused recent cholera outbreaks in India during the past years. Phage sensitivity data propose an alternative cost-effective approach to prevent cholera outbreak by therapeutic uses of typing phages irrespective of origin or clonality of the strains.

Representatives of the genus Vibrio cholerae differ in the structure of lipopolysaccharide, in particular, its O-polysaccharide chains (O-antigen), which determines the serological specificity of vibrios. Currently, the water-phenolic method is used to obtain the lipopolysaccharide preparation. However, this technique relates to harsh chemical methods, leads to a change in original molecular organization of biopolymer, violating its structure and biological properties. Modern technologies in the development of diagnostic kits for the immunosuspension reaction of volume agglomeration allow for obtaining synthetic carriers with different reaction groups on the particle surface capable to bind antigens/antibodies. The aim of this study was to construct cholera antigenic polymeric diagnostic kit based on the lipopolysaccharide of Vibrio cholerae O1 serogroup. Materials and methods. The lipopolysaccharide was used as a sensitizer obtained through the author's modification of enzymatic purification from the cell membranes of Vibrio cholerae using ultrasonic disintegration. Results and discussion. The resulting sensitin contains small impurities of protein (1.5 %) and nucleic acids (0.1 %). Diagnosticum is characterized by high analytical sensitivity in agglomeration reaction with commercial and experimental rabbit serum to Vibrio cholerae O1 serogroup (1:640 - 1:5120) and analytical specificity (the diagnosticum does not interact with heterologous sera, with serums to pathogens of acute intestinal infections, as well as with sera from healthy donors). A polymeric antigenic cholera diagnosticum designed to detect antibodies to lipopolysaccharide of Vibrio cholerae in the blood serum of patients who were ill, suspected of the disease or vaccinated people has been constructed.

Background & objectives: Antibiotic resistance in Vibrio cholerae is a crucial matter in the world. Objective of this study was the improvement of cholera surveillance by assessing the antimicrobial resistance pattern and bacterial resistance genes in V. cholerae O1 isolates, reffered to Iranian Reference Health Laboratory, in cholera outbreaks during 2012- 2015. Methods: This study is a cross sectional- descriptive research. Antimicrobial susceptibility test (AST) to 8 antibiotics was performed on 113 V.cholerae O1 isolates using E-test method. For all isolates, conventional PCR method was used to detect the presence of tetracycline resistance genes (tetA, tetB and tetC) and the sulfamethoxazole-trimethoprim resistance genes (sul2 and dfrA1). Results: All isolates were sensitive to ampicillin, temocillin, ciprofloxacin and cefixime and 64% of strains showed intermediate susceptibility to erythromycin. The resistance rate of nalidixic acid, sulfamethoxazole-trimethoprim and tetracycline were 90%, 71% and 50% respectively. However, the frequency of multidrug resistant (MDR) strains varied across the years. The frequency of resistance genes (tetA, tetB, tetC, sul2 and dfrA1) were 70%, 34%, 58%, 66% and 70% respectively. Conclusion: AST should be used to determine the resistance profile at the beginning of a cholera outbreak and to monitor the resistance profile of circulating strains as part of surveillance of the disease. A prominent association was observed between phenotypic resistance to sulfamethoxazole-trimethoprim and presence of dfrA1gene. Determining the presence of resistance genes is necessary for understanding the epidemiology and routes of transmission of antibiotic resistance genes

Introduction: Vibrio cholera is a gram-negative bacterium that causes cholera which is an acute diarrheal illness. Cholera outbreak mostly happens in hot seasons when the bacteria reproduce rapidly in water contaminated with feces. Severe diarrhea and mortality have been the prevalent symptoms of cholera which affect the hygienic status and economic features of humans during centuries. There is a challenge currently to plan and propose a quick and reliable method to detect vibrio cholera which has been one of the fast growing and contagious disease in this decade. Materials & Methods: New Zealand white rabbits were immunized with vibrio cholera Ogawa and Inaba whole cells. The obtained gamma globulin from the whole serum was purified through ammonium sulfate precipitation method and then concentrated in this study. Subsequently, the purity of these antigens increased with the use of mixed mode-chromatography in the next step. The purified antibodies were connected to staphylococcus aureus cowan 1 NCTC-8325 whole cells. Rectal swab samples were inoculated into an enriched Alkaline Peptone Water medium (pH 8.6) incubated for 5 h at 37 °C. One drop from each sample was mixed with one drop of vibrio cholera co-agglutination complex. Subsequently, the results were read in 2-3 min. Findings: The obtained results showed 97%, 99%, and 98% of sensitivity, specificity, and accuracy from the proposed assay, respectively, compared to the standard culture methods. This proposed assay is considered as a rapid and reliable method to evaluate endemic or epidemic transmission of fatal illnesses. Discussion & Conclusions: The proposed co-agglutination test is a simple, rapid, and reliable method to detect Vibrio cholera in fecal specimens. This method can be utilized to detect and control severe diarrhea which results from vibrio leading to the prevention of mortality, especially in areas with minimal laboratory facilities.

Objective: Cholera is an acute secretory diarrhea caused by the Gram-negative bacterium, Vibrio cholerae mostly through production of cholera toxin (CT) and zonula occludens toxin (Zot). Isolates of V. cholerae have acquired resistance elements during the last decade. One of the most promising ways to treat resistant strains is to use antivirulence agents instead of killing the causative agent with conventional antibiotics. In this study, we examined whether different concentrations of capsaicin - the pungent fraction of red chili- can act as an antivirulence agent and inhibit V. cholerae toxin production. Materials and Methods: Two standard strains namely, V. cholerae ATCC 14035 and V. cholerae PTCC 1611 were used in this study. Minimum Inhibitory Concentration (MIC) of capsaicin was determined by broth microdilution method. Based on MIC results, the bacteria were cultured in the presence of sub-MIC concentrations of capsaicin and a negative control without capsaicin. Real-time PCR (RT-PCR) was carried out to determine the expression level of V. cholerae toxin genes at each concentration. Results: MIC test showed that 200 mg/mL of capsaicin in 2% dimethyl sulfoxide (DMSO) could inhibit the growth of the two standard strains of V. cholerae. The expression of V. cholerae toxin genes was significantly reduced following treatment with sub-MIC concentrations of capsaicin as assessed by RT-PCR. Conclusion: Capsaicin showed great inhibitory effect against cholera toxin and reduced Zot production in the tested strains of V. cholerae. The results showed promising insights into antivirulence effects of capsaicin.

A series of sixteen benzenesulfonamide derivatives has been synthesised and tested as inhibitors of Vibrio cholerae carbonic anhydrase (CA) enzymes, belonging to α-CA, β-CA, and γ-CA classes (VchCAα, VchCAβ, and VchCAγ). The determined Ki values were compared to those of selected human CA isoforms (hCA I and hCA II). Structure-affinity relationship analysis highlighted that all tested compounds proved to be active inhibitors of VchCAα at nanomolar concentration. The VchCAβ activity was lower to respect inhibitory efficacy toward VchCAα, whereas, these benzenesulfonamide derivatives failed to inhibit VchCAγ. Interestingly, compound 7e combined the best activity toward VchCAα and VchCAβ. In order to obtain a model for binding mode of our inhibitors toward bacterial CAs, we carried out docking simulations by using the available crystal structures of VchCAβ.

Cholera is one of the most important epidemic diseases globally. It is causing of morbidity and mortality in the world. Severe watery diarrhea causes a composite process involving several component that help them reach the lining of the small intestine, form colonies and generate bacterial toxins. This survey was carried out to discover the genetic patterns of clinical isolates according to the presence or absence of toxic genes for the city of Diwaniyah in southern Iraq. Sixty isolates were isolated from patients with cholera. The isolates included the center of the city and the surrounding rural areas. Biochemical and serological diagnosis. All the isolates were V. cholera serogroup O 1 of the serotyping Ogawa in biotype El Tor. Genetic testing was carried out using PCR technique and base on the presence or absence of toxin genes. Three genotypes were identified for the region.

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Background: Susceptibility to Vibrio cholerae infection is affected by blood group, age, and preexisting immunity, but these factors only partially explain who becomes infected. A recent study used 16S ribosomal RNA amplicon sequencing to quantify the composition of the gut microbiome and identify predictive biomarkers of infection with limited taxonomic resolution.Methods: To achieve increased resolution of gut microbial factors associated with V. cholerae susceptibility and identify predictors of symptomatic disease, we applied deep shotgun metagenomic sequencing to a cohort of household contacts of patients with cholera.Results: Using machine learning, we resolved species, strains, gene families, and cellular pathways in the microbiome at the time of exposure to V. cholerae to identify markers that predict infection and symptoms. Use of metagenomic features improved the precision and accuracy of prediction relative to 16S sequencing. We also predicted disease severity, although with greater uncertainty than our infection prediction. Species within the genera Prevotella and Bifidobacterium predicted protection from infection, and genes involved in iron metabolism were also correlated with protection.Conclusion: Our results highlight the power of metagenomics to predict disease outcomes and suggest specific species and genes for experimental testing to investigate mechanisms of microbiome-related protection from cholera. [ABSTRACT FROM AUTHOR]

The study included many parts; the first part was the isolation and identification of Vibrio cholerae from diarrhea cases occurred in Al-Muthanna province, 150 diarrhea samples were collected from hospitalized patients in Al-Hussein Teaching Hospital, Women's and Children's Teaching Hospital, Al-Rumaytha General Hospital and Al-Khudher Hospital. The samples were cultured directly on Alkaline peptone water (APW), TCBS agar and Blood agar then the vibrio was identified by microscopic and biochemical tests as well as APl20E was used to confirm identification. The serotype was identified by antisera in addition to biochemical tests used to differentiate the biotype EI TOR and Classical. Spring test was used to differentiate Vibrio spp. from Aeromonas spp. The second part included statistical study of Cholerae epidemiology in Al-Muthanna province during the years of 2015, 2016, 2017 and 2018 in order to identify patient numbers and the most common serotypes. The third part was to identify the antibiotic susceptibility profile of V. colerae isolates against 12 antibiotics (clindamycine, tetracycline, pipracilline, Cefoxitin, amikacin, ampicillin, amoxicillin clavanic acid, nitrofurantoin, ticarcillin-clavulanic acid, erythromycin, nalidic acid and cefapim). The phenotypic characteristics, biochemical test, APl20E, and VITEK SYS revealed that only 10 isolates 6.66% out of 150 diarrhea sample were V. cholerae O1 EI TOR of INABA serotype. The epidemiological survey uncovered that the year of 2015 was the higher year recording cholerae infection at 294 infections in comparison to other years while no cholerae infection was recorded during the 2016. INABA serotype was the most common serotype and the infections were recorded in the males more than women at statistical significant (p< 0.05). The results showed that Samawa city/Al- Muthanna province was higher city of infection cases during 2015, 2017, and 2018 compared to other cities, where The antibiotic susceptibility profile showed that all V. cholerae isolates were 100% sensitive to Amikacin, Nalidic acid, cifampim and Nitrofurantoin while 100% resistant to Amoxicillin, Erythromycin, ticarcillin-clavulanic acid, Cefoxitin, ampicillin, clindamycin and Tetracyclin while were moderate to pipracilline. [ABSTRACT FROM AUTHOR]

Background: Cholera is a primordial disease caused by Vibrio cholerae which existed from centuries in different parts of the world and still shows its periodic, endemic and epidemic presence. Thousands of cholera cases are reported from different parts of India and the disease remains endemic throughout the year. At present, we do not have enough knowledge about the phenotypic nature of the circulating V. cholerae strains in this part of the world. Objectives: This study was carried out over a period of 6 years with the aim defer with the changes in the prevalence and distribution of biotypes, serotypes and phage types of V. cholerae clinical isolates from various endemic regions of the country to determine phenotypic characteristics of the circulating strains and also to predict the attributes of cholera strains responsible for causing significant outbreaks in future. Materials and Methods: A total of 1882 V.cholerae O1 isolates from different cholera endemic areas of India were included in this study. V.cholerae strains which were identified as O1 biotype ElTor further analyzed for serotype and phage types using the standard methodologies. Polyvalent O1 and monospecific Inaba and Ogawa antisera were used for serotyping. A panel of five phages of Basu and Mukherjee phage typing scheme and five phages from the new phage typing scheme were used for phage typing analysis following standard methodology. Results: Maximum numbers of strains were isolated from cholera-endemic states like Gujarat and Maharashtra. All the isolates were confirmed as V. cholerae O1 biotype ElTor and majority of them were serotype Ogawa (93.2%). New phage typing scheme resulted in almost 100% typeable V. cholerae O1 strains included in this study and phage type 27 was the predominant type. Although 80% of the strains used in this study were sensitive to all the vibrio phages, S5 phage was found most efficient in lysing cholera strains indicating its broader host range. Conclusion: The current study identified phage type 27 as the most dominant type and serotype Ogawa was found continuous in circulation throughout the year which has caused recent cholera outbreaks in India during the past years. Phage sensitivity data propose an alternative cost-effective approach to prevent cholera outbreak by therapeutic uses of typing phages irrespective of origin or clonality of the strains. [ABSTRACT FROM AUTHOR]

Cholera infections caused by the gamma-proteobacterium Vibrio cholerae have ravaged human populations for centuries, and cholera pandemics have afflicted every corner of the globe. Fortunately, interventions such as oral rehydration therapy, antibiotics/antimicrobials, and vaccines have saved countless people afflicted with cholera, and new interventions such as probiotics and phage therapy are being developed as promising approaches to treat even more cholera infections. Although current therapies are mostly effective and can reduce disease transmission, cholera outbreaks remain deadly, as was seen during recent outbreaks in Haiti, Ethiopia, and Yemen. This is due to significant underlying political and socioeconomic complications, including shortages of vaccines and clean food and water and a lack of health surveillance. In this review, we highlight the strengths and weaknesses of current cholera therapies, discuss emerging technologies, and argue that a multi-pronged, flexible approach is needed to continue to reduce the worldwide burden of cholera.

Abstract Background A nationwide outbreak of Vibrio cholerae occurred in Ghana in 2014 with Accra, the nation’s capital as the epi-center. The outbreak spread to the Brong Ahafo Region (BAR) which is geographically located in the middle of the country. In this region a review of data collected during the outbreak was carried out and analyzed descriptively to determine the hot spots and make recommendations for effective response to future outbreaks. Methods A review of patient records and line lists of cases of cholera reported in all hospitals during the period of the outbreak (July–December 2014) was conducted. Hospitals used IDSR (Integrated Disease Surveillance and Response system) standard case definitions to detect and report cases for management. The GPS coordinates of all districts and health facilities were collected and utilized in the construction of spot maps. We also obtained populations (denominators) from the BAR Health surveillance unit of the Ghana Health Service. All the data thus collected was analyzed descriptively and expressed as frequencies and rates. Results A total of 1035 cases were reported, 550 (53.4%) were males and the rest females. Their ages ranged from 1 to 95 years; (mean age of 28.2 ± 19.6 years). The most affected (23.5%) was the 20–29 year old age group. On the 30th July, 2014, a 26 year old male (recorded as the index case of the cholera outbreak in the Brong Ahafo region) with a history of travel from Accra reported to the Nkoranza district hospital with a history of symptoms suggestive of cholera. The reporting of cholera cases reached their peak (17.3%) in week 15 of the outbreak (this lasted 25 weeks). An overall attack rate of 71/100,000 population, and a case fatality rate of 2.4% was recorded in the region. Asutifi South district however recorded a case fatality of 9.1%, the highest amongst all the districts which recorded outbreaks. The majority of the cases reported in the region were from Atebubu-Amanten, Sene West, Pru, and Asunafo North districts with 31.1, 26.0, 18.2 and 9.9% respectively. Vibrio cholerae serotype O1 was isolated from rectal swabs/stool samples tested. Conclusion Vibrio cholerae serotype O1 caused the cholera-outbreak in the Brong Ahafo Region and mainly affected young adult-males. The most affected districts were Atebubu-Amanten, Sene west, Pru (located in the eastern part of the region), and Asunafo North districts (located in the south west of the region). Case Fatality Rate was higher (2.4%) than the WHO recommended rate (

A series of 2-thio- and 2-seleno-acetamides bearing the benzenesulfonamide moiety were evaluated as Carbonic Anhydrase (CA, EC 4.2.1.1) inhibitors against different pathogenic bacteria such as the Vibrio cholerae (VchCA-α and VchCA-β), Burkholderia pseudomallei (BpsCA-β and BpsCA-β ), Mycobacterium tuberculosis (Rv3723-β) and the Salmonella enterica serovar Typhimurium (StCA2-β). The molecules represent interesting leads worth developing as innovative antibacterial agents since they possess new mechanism of action and isoform selectivity preferentially against the bacterial expressed CAs. The identification of potent and selective inhibitors of bacterial CAs may lead to tools also useful for deciphering the physiological role(s) of such proteins. [ABSTRACT FROM AUTHOR]

A series of sixteen benzenesulfonamide derivatives has been synthesised and tested as inhibitors of Vibrio cholerae carbonic anhydrase (CA) enzymes, belonging to α-CA, β-CA, and γ-CA classes (VchCAα, VchCAβ, and VchCAγ). The determined Ki values were compared to those of selected human CA isoforms (hCA I and hCA II). Structure-affinity relationship analysis highlighted that all tested compounds proved to be active inhibitors of VchCAα at nanomolar concentration. The VchCAβ activity was lower to respect inhibitory efficacy toward VchCAα, whereas, these benzenesulfonamide derivatives failed to inhibit VchCAγ. Interestingly, compound 7e combined the best activity toward VchCAα and VchCAβ. In order to obtain a model for binding mode of our inhibitors toward bacterial CAs, we carried out docking simulations by using the available crystal structures of VchCAβ. [ABSTRACT FROM AUTHOR]

The de novo pyrimidine biosynthesis pathway is essential for the proliferation of many pathogens. One of the pathway enzymes, dihydroorotase (DHO), catalyzes the reversible interconversion of N-carbamoyl- l -aspartate to 4,5-dihydroorotate. The substantial difference between bacterial and mammalian DHOs makes it a promising drug target for disrupting bacterial growth and thus an important candidate to evaluate as a response to antimicrobial resistance on a molecular level. Here, we present two novel three-dimensional structures of DHOs from Yersinia pestis (Yp DHO), the plague-causing pathogen, and Vibrio cholerae (Vc DHO), the causative agent of cholera. The evaluations of these two structures led to an analysis of all available DHO structures and their classification into known DHO types. Comparison of all the DHO active sites containing ligands that are listed in DrugBank was facilitated by a new interactive, structure-comparison and presentation platform. In addition, we examined the genetic context of characterized DHOs, which revealed characteristic patterns for different types of DHOs. We also generated a homology model for DHO from Plasmodium falciparum. Unlabelled Image [ABSTRACT FROM AUTHOR]

Objective: Cholera is an acute secretory diarrhea caused by the Gram-negative bacterium, Vibrio cholerae mostly through production of cholera toxin (CT) and zonula occludens toxin (Zot). Isolates of V. cholerae have acquired resistance elements during the last decade. One of the most promising ways to treat resistant strains is to use antivirulence agents instead of killing the causative agent with conventional antibiotics. In this study, we examined whether different concentrations of capsaicin - the pungent fraction of red chili-can act as an antivirulence agent and inhibit V. cholerae toxin production. Materials and Methods: Two standard strains namely, V. cholerae ATCC 14035 and V. cholerae PTCC 1611 were used in this study. Minimum Inhibitory Concentration (MIC) of capsaicin was determined by broth microdilution method. Based on MIC results, the bacteria were cultured in the presence of sub-MIC concentrations of capsaicin and a negative control without capsaicin. Real-time PCR (RT-PCR) was carried out to determine the expression level of V. cholerae toxin genes at each concentration. Results: MIC test showed that 200 mg/mL of capsaicin in 2% dimethyl sulfoxide (DMSO) could inhibit the growth of the two standard strains of V. cholerae. The expression of V. cholerae toxin genes was significantly reduced following treatment with sub-MIC concentrations of capsaicin as assessed by RT-PCR. Conclusion: Capsaicin showed great inhibitory effect against cholera toxin and reduced Zot production in the tested strains of V. cholerae. The results showed promising insights into antivirulence effects of capsaicin. [ABSTRACT FROM AUTHOR]

Cholera remains a major risk in developing countries, particularly after natural or man-made disasters. Vibrio cholerae El Tor is the most important cause of these outbreaks, and is becoming increasingly resistant to antibiotics, so alternative therapies are urgently needed. In this study, a single bacteriophage, Phi_1, was used to control cholera prophylactically and therapeutically in an infant rabbit model. In both cases, phage-treated animals showed no clinical signs of disease, compared with 69% of untreated control animals. Bacterial counts in the intestines of phage-treated animals were reduced by up to 4 log10 colony-forming units/g. There was evidence of phage multiplication only in animals that received a V. cholerae challenge. No phage-resistant bacterial mutants were isolated from the animals, despite extensive searching. This is the first evidence that a single phage could be effective in the treatment of cholera, without detectable levels of resistance. Clinical trials in human patients should be considered. [ABSTRACT FROM AUTHOR]