RESUMEN
Sensing and resisting oxidative stress is critical for Vibrio cholerae to survive in either the aquatic environment or the gastrointestinal tract. Previous studies mainly focused on the mechanisms of oxidative stress response regulation that rely on enzymatic antioxidant systems, while functions of non-enzymatic antioxidants are rarely discussed in V. cholerae. For the first time, we investigated the role of hydrogen sulfide (H2S), the simplest thiol compound, in protecting V. cholerae against oxidative stress. We found that degradation of L-cysteine by putative cystathionine ß-synthase (CBS) is the major source of endogenous H2S in V. cholerae. Our results indicate that intracellular H2S level has a positive correlation with cbs expression, while the enhanced H2S production can render V. cholerae cells less susceptible to H2O2 in vitro. Using proteome analysis and real-time qPCR assay, we found that cbs expression could stimulate the expression of several enzymatic antioxidants, including reactive oxygen species (ROS) detoxifying enzymes SodB, KatG and AhpC, the DNA protective protein DPS and the protein redox regulator Trx1. Assays of ROS detoxification capacities revealed that CBS-derived H2S could promote catalase activity at the post-translational level, especially for KatB, which serves as an important way that endogenous H2S participates in H2O2 detoxification. The enhancement of catalase activity by H2S is achieved through facilitating the uptake of iron. Adult mice experiments showed that cbs mutant has colonization defect, while either complementation of cbs or exogenous supplement of N-Acetyl-L-Cysteine restores its fitness in the host environment. Herein, we proposed that V. cholerae regulates CBS-dependent H2S production for better survival and proliferation under ROS stress.
Asunto(s)
Cistationina betasintasa/metabolismo , Interacciones Huésped-Patógeno/fisiología , Sulfuro de Hidrógeno/metabolismo , Cinesinas/metabolismo , Vibrio cholerae/metabolismo , Animales , Proteínas Bacterianas/metabolismo , Catalasa/metabolismo , Cólera/metabolismo , Ratones , Estrés Oxidativo/fisiología , Vibrio cholerae/patogenicidadRESUMEN
The use of traditional foods and beverages or their bioactive compounds as anti-virulence agents is a new alternative method to overcome the increased global emergence of antimicrobial resistance in enteric pathogens. In the present study, we investigated the anti-virulence activity of a polyphenolic fraction previously isolated from Kombucha, a 14-day fermented beverage of sugared black tea, against Vibrio cholerae O1. The isolated fraction was mainly composed of the polyphenols catechin and isorhamnetin. The fraction, the individual polyphenols and the combination of the individual polyphenols significantly inhibited bacterial swarming motility and expression of flagellar regulatory genes motY and flaC, even at sub-inhibitory concentrations. The polyphenolic compounds also decreased bacterial protease secretion and mucin penetration in vitro. In vivo study revealed that the polyphenolic fraction significantly inhibited V. cholerae induced fluid accumulation in the rabbit ileal loop model and intestinal colonization in suckling mice model. Therefore, the anti-virulence activity of the Kombucha polyphenolic fraction involved inhibition of motility and protease secretion of V. cholerae, thus preventing bacterial penetration through the mucin layer as well as fluid accumulation and bacterial colonization in the intestinal epithelial cells. The overall results implied that Kombucha might be considered as a potential alternative source of anti-virulence polyphenols against V. cholerae. To the best of our knowledge, this is the first report on the anti-virulence activity of Kombucha, mostly attributed to its polyphenolic content.
Asunto(s)
Té de Kombucha , Polifenoles/farmacología , Vibrio cholerae/efectos de los fármacos , Animales , Antibacterianos/farmacología , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/metabolismo , Proteínas Bacterianas/efectos de los fármacos , Catequina/farmacología , Movimiento Celular/efectos de los fármacos , Cólera/tratamiento farmacológico , Expresión Génica/efectos de los fármacos , Intestino Delgado/efectos de los fármacos , Intestino Delgado/microbiología , Ratones , Péptido Hidrolasas/efectos de los fármacos , Extractos Vegetales/farmacología , Quercetina/análogos & derivados , Quercetina/farmacología , Conejos , Vibrio cholerae/patogenicidad , Virulencia/efectos de los fármacos , Virulencia/genética , Factores de Virulencia/genética , Factores de Virulencia/metabolismoRESUMEN
Acute Vibrio cholerae infection triggers significant inflammatory response and immense fluid secretion in the intestine. In the present study, methyl gallate (MG) isolated from Terminalia chebula was evaluated to determine the in vivo fluid accumulation-inhibitory, anticolonization and anti-inflammatory and in vitro biofilm-inhibitory activities against multi-drug resistant (MDR) V. cholerae. Bacterial membrane-damaging and biofilm-inhibitory activities were determined by membrane perturbation and transmission electron microscopy (TEM); and microdilution assays, respectively. Fluid accumulation-inhibitory and anticolonization activities of MG (23.80-95.23â¯mg/kg body weight) were determined in 4-5 days old BALB/c mice with an incubation time of 18â¯h. The effect of MG (1, 50 and 500â¯mg/kg body weight) on intestinal inflammatory reaction induced by V. cholerae was studied by performing histology in Swiss albino mice. MIC and MBC of MG against the test strains were 32-64 and 64-256⯵g/ml, respectively. MG showed the fluid accumulation-inhibitory activity with inhibition values of 42.86-89.08% at doses between 23.80 and 95.23â¯mg/kg body weight and significant anticolonization activity (pâ¯<â¯0.0001) against V. choleare in the suckling mouse intestine. MG (500â¯mg/kg body weight) significantly inhibited the inflammatory reactions induced by V. cholerae compared to the vehicle control. MG exhibited 70% minimum biofilm inhibition concentration of 64⯵g/ml and bacterial membrane damaging activity at 1â¯×â¯MBC. The results obtained in the present study suggest that MG has potential as an effective agent for the treatment of severe secretory and inflammatory diarrheal disease caused by MDR V. cholerae.
Asunto(s)
Antibacterianos/farmacología , Antiinflamatorios/farmacología , Biopelículas/efectos de los fármacos , Fluoroquinolonas/farmacología , Ácido Gálico/análogos & derivados , Terminalia/química , Vibrio cholerae/efectos de los fármacos , Animales , Antibacterianos/aislamiento & purificación , Membrana Celular/efectos de los fármacos , Cólera/microbiología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Ácido Gálico/administración & dosificación , Ácido Gálico/aislamiento & purificación , Ácido Gálico/farmacología , Intestino Delgado/patología , Intestino Delgado/virología , Ratones , Ratones Endogámicos BALB C , Extractos Vegetales/farmacología , Vibrio cholerae/citología , Vibrio cholerae/crecimiento & desarrollo , Vibrio cholerae/patogenicidadRESUMEN
Antibiotics are important adjuncts to oral rehydration therapy in cholera disease management. However, due to the rapid emergence of resistance to the antibiotics used to treat cholera, therapeutic options are becoming limited. Therefore, there is a critical need to develop additional therapeutics to aid in the treatment of cholera. Previous studies showed that the extracytoplasmic stress response (σE) pathway of Vibrio cholerae is required for full virulence of the organism. The pathway is also required for bacterial growth in the presence of ethanol. Therefore, we exploited this ethanol sensitivity phenotype in order to develop a screen for inhibitors of the pathway, with the aim of also inhibiting virulence of the pathogen. Here we describe the optimization and implementation of our high-throughput screening strategy. From a primary screen of over 100,000 compounds, we have identified seven compounds that validated the growth phenotypes from the primary and counterscreens. These compounds have the potential to be developed into therapeutic agents for cholera and will also be valuable probes for uncovering basic molecular mechanisms of an important cause of diarrheal disease.
Asunto(s)
Evaluación Preclínica de Medicamentos/métodos , Ensayos Analíticos de Alto Rendimiento/métodos , Vibrio cholerae/efectos de los fármacos , Vibrio cholerae/patogenicidad , VirulenciaRESUMEN
Orthologs search identified that the Vibrio cholerae gluconate (Gnt) utilization system minimally consisted of the Entner-Doudoroff (ED) pathway (edd and eda) and three other genes, namely gntU, gntK and gntR This system appeared unique by genomic organization of component genes into two operons transcribed in opposite directions. In silico analysis indicated GntU as an inner-membrane protein functioning for transport and GntK as a kinase with cytosolic localization that generates Gnt6P, which is then metabolized through the ED pathway. Enzyme 6-phosphogluconate dehydratase encoded by edd converts Gnt6P to 2-keto-3-deoxy-6-phosphogluconate (KDPG), which is metabolized by the action of KDPG-aldolase encoded by eda Transcriptional upregulation of the Gnt utilization genes in the gntR mutant matched well to a predicted repressor role of GntR. GntR displayed DNA binding to a region in the promoters of two bi-directionally transcribed operons. Growth defect of mutants in Gnt-supplemented media confirmed obligate involvement of these genes in Gnt utilization and such defect was restored upon complementation. Defective Gnt utilization resulted in attenuation of colonization potential and reduction of cholera toxin secretion in V. cholerae The ED pathway mutants showed the highest level of virulence attenuation. Overall, this study established a minimal requirement of the V. cholerae Gnt utilization system, which played a critical role in pathogenesis.
Asunto(s)
Gluconatos/metabolismo , Vibrio cholerae/fisiología , Secuencia de Aminoácidos , Animales , Cólera/microbiología , Orden Génico , Genes Bacterianos , Proteínas de Transporte de Membrana/química , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Redes y Vías Metabólicas , Mutación , Operón , Conejos , Vibrio cholerae/patogenicidad , Virulencia/genéticaRESUMEN
Nanomaterial-based photoluminescence (PL) diagnostic devices offer fast and highly sensitive detection of pesticides, DNA, and toxic agents. Here we report a label-free PL genosensor for sensitive detection of Vibrio cholerae that is based on a DNA hybridization strategy utilizing nanostructured magnesium oxide (nMgO; size >30 nm) particles. The morphology and size of the synthesized nMgO were determined by transmission electron microscopic (TEM) studies. The probe DNA (pDNA) was conjugated with nMgO and characterized by X-ray photoelectron and Fourier transform infrared spectroscopic techniques. The target complementary genomic DNA (cDNA) isolated from clinical samples of V. cholerae was subjected to DNA hybridization studies using the pDNA-nMgO complex and detection of the cDNA was accomplished by measuring changes in PL intensity. The PL peak intensity measured at 700 nm (red emission) increases with the increase in cDNA concentration. A linear range of response in the developed PL genosensor was observed from 100 to 500 ng/µL with a sensitivity of 1.306 emi/ng, detection limit of 3.133 ng/µL and a regression coefficient (R(2)) of 0.987. These results show that this ultrasensitive PL genosensor has the potential for applications in the clinical diagnosis of cholera.
Asunto(s)
Técnicas Biosensibles , Cólera/diagnóstico , ADN Bacteriano/aislamiento & purificación , Óxido de Magnesio/química , Nanoestructuras/química , Vibrio cholerae/aislamiento & purificación , Cólera/microbiología , Cólera/patología , Sondas de ADN/síntesis química , Sondas de ADN/química , ADN Bacteriano/genética , ADN Complementario/química , ADN Complementario/genética , Humanos , Límite de Detección , Mediciones Luminiscentes , Nanoestructuras/ultraestructura , Hibridación de Ácido Nucleico/métodos , Procesos Fotoquímicos , Espectroscopía Infrarroja por Transformada de Fourier , Vibrio cholerae/genética , Vibrio cholerae/patogenicidadRESUMEN
The severe diarrheal disease cholera is endemic in over 50 countries. Current therapies for cholera patients involve oral and/or intravenous rehydration, often combined with the use of antibiotics to shorten the duration and intensity of the disease. However, as antibiotic resistance increases, treatment options will become limited. Linoleic acid has been shown to be a potent negative effector of V. cholerae virulence that acts on the major virulence transcription regulator protein, ToxT, to inhibit virulence gene expression. ToxT activates transcription of the two major virulence factors required for disease, cholera toxin (CT) and toxin-coregulated pilus (TCP). A conjugated form of linoleic acid (CLA) is currently sold over the counter as a dietary supplement and is generally recognized as safe by the U.S. Food and Drug Administration. This study examined whether CLA could be used as a new therapy to reduce CT production, which, in turn, would decrease disease duration and intensity in cholera patients. CLA could be used in place of traditional antibiotics and would be very unlikely to generate resistance, as it affects only virulence factor production and not bacterial growth or survival.
Asunto(s)
Antibacterianos/farmacología , Proteínas Bacterianas/antagonistas & inhibidores , Toxina del Cólera/biosíntesis , Ácidos Linoleicos Conjugados/farmacología , Factores de Transcripción/antagonistas & inhibidores , Vibrio cholerae/efectos de los fármacos , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Cólera/tratamiento farmacológico , Cólera/fisiopatología , Toxina del Cólera/genética , ADN Bacteriano/metabolismo , Modelos Animales de Enfermedad , Regulación Bacteriana de la Expresión Génica , Conejos , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Vibrio cholerae/metabolismo , Vibrio cholerae/patogenicidad , Virulencia/genética , Factores de Virulencia/genética , Factores de Virulencia/metabolismoRESUMEN
Botanicals are rich in bioactive compounds, and some offer numerous beneficial effects to animal and human health when consumed. It is well known that phytochemicals in cranberries have anti-oxidative and antimicrobial activities. Recently, an increasing body of evidence has demonstrated that cranberry phytochemicals may have potential benefits that promote healthy aging. Here, we use Caenorhabditis elegans as a model to show that water-soluble cranberry extract standardized to 4.0% proanthocyanidins (WCESP), a major component of cranberries, can enhance host innate immunity to resist against Vibrio cholerae (V. cholerae; wild type C6706 (O1 El Tor biotype)) infection. Supplementation of WCESP did not significantly alter the intestinal colonization of V. cholerae, but upregulated the expression of C. elegans innate immune genes, such as clec-46, clec-71, fmo-2, pqn-5 and C23G10.1. Additionally, WCESP treatment did not affect the growth of V. cholerae and expression of the major bacterial virulence genes, and only slightly reduced bacterial colonization within C. elegans intestine. These findings indicate that the major components of WCESP, including proanthocyanidins (PACs), may play an important role in enhancing the host innate immunity. Moreover, we engaged C. elegans mutants and identified that the p38 MAPK signaling, insulin/IGF-1 signaling (IIS), and HSF-1 play pivotal roles in the WCESP-mediated host immune response. Considering the level of conservation between the innate immune pathways of C. elegans and humans, the results of this study suggest that WCESP may also play an immunity-promoting role in higher order organisms.
Asunto(s)
Proteínas de Caenorhabditis elegans/biosíntesis , Inmunidad Innata/efectos de los fármacos , Extractos Vegetales/administración & dosificación , Proantocianidinas/administración & dosificación , Factores de Transcripción/biosíntesis , Proteínas Quinasas p38 Activadas por Mitógenos/biosíntesis , Envejecimiento/efectos de los fármacos , Envejecimiento/genética , Animales , Caenorhabditis elegans/efectos de los fármacos , Caenorhabditis elegans/genética , Caenorhabditis elegans/inmunología , Proteínas de Caenorhabditis elegans/genética , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Factor I del Crecimiento Similar a la Insulina/biosíntesis , Extractos Vegetales/química , Proantocianidinas/química , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/genética , Vaccinium macrocarpon/química , Vibrio cholerae/efectos de los fármacos , Vibrio cholerae/patogenicidad , Proteínas Quinasas p38 Activadas por Mitógenos/genéticaRESUMEN
Resveratrol, a phytochemical commonly found in the skin of grapes and berries, was tested for its biofilm inhibitory activity against Vibrio cholerae. Biofilm inhibition was assessed using crystal violet assay. MTT assay was performed to check the viability of the treated bacterial cells and the biofilm architecture was analysed using confocal laser scanning microscopy. The possible target of the compound was determined by docking analysis. Results showed that subinhibitory concentrations of the compound could significantly inhibit biofilm formation in V. cholerae in a concentration-dependent manner. AphB was found to be the putative target of resveratrol using docking analysis. The results generated in this study proved that resveratrol is a potent biofilm inhibitor of V. cholerae and can be used as a novel therapeutic agent against cholera. To our knowledge, this is the first report of resveratrol showing antibiofilm activity against V. cholerae.
Asunto(s)
Biopelículas/efectos de los fármacos , Extractos Vegetales/farmacología , Estilbenos/farmacología , Vibriosis/microbiología , Vibrio cholerae/patogenicidad , Vitis/química , Proteínas Bacterianas/metabolismo , Relación Dosis-Respuesta a Droga , Simulación del Acoplamiento Molecular , Resveratrol , Estilbenos/uso terapéutico , Transactivadores/metabolismo , Vibriosis/tratamiento farmacológico , Vibrio cholerae/metabolismoRESUMEN
AIM: Study of plant extracts that have the ability to neutralize cytotoxic activity of hemolysin. MATERIALS AND METHODS: Preparations of purified and recombinant V. cholerae eltor hemolysin as well as supernatants of V. cholerae strains were used. Determination ofcytotoxic activity of hemolysin and neutralizing activity of plant extracts were carried out by using cell cultures CHO-K1 and CaCo2. RESULTS: Out of 9 water extracts only 3 - extracts of Rhei rhizome, Limonium gmelinii and Quercus robur neutralized hemolysin in cell culture CHO-K1 and CaCo2, whereas the other extracts--Humulus lupulus, Ocimum basilicum, Chelidonium majus, Juglans regia, Achillea milefolium and Hypericum perforatum did not have anti-cytotoxic effect. Neutralizing properties of extracts are exhibited during their co-incubation with hemolysin preparations and supernatants of V. cholerae strains already within 10 minutes. CONCLUSION: Plant extracts that have anti-cytotoxic activity against hemolysin are perspective for development oftherapeutic-prophylaxis preparations.
Asunto(s)
Proteínas Bacterianas/efectos de los fármacos , Cólera/tratamiento farmacológico , Proteínas Hemolisinas/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Proteínas Bacterianas/toxicidad , Células CHO , Células CACO-2 , Cólera/microbiología , Toxina del Cólera/antagonistas & inhibidores , Toxina del Cólera/química , Cricetinae , Cricetulus , Proteínas Hemolisinas/toxicidad , Humanos , Extractos Vegetales/química , Plumbaginaceae/química , Quercus/química , Vibrio cholerae/efectos de los fármacos , Vibrio cholerae/patogenicidadRESUMEN
The Entner-Doudoroff (ED) pathway has recently been shown to play an important role in sugar catabolism for many organisms although very little information is available on the functionality of this pathway in Vibrio cholerae, the causative agent of cholera. In this study, activation of the genes edd and eda, encoding 6-phosphogluconate dehydratase and 2-keto-3-deoxy-6-phosphogluconate aldolase, was used as a marker of a functional ED pathway in V. cholerae. Transcriptional activation analyses and gene silencing experiments with cells grown in sugar-supplemented M9 medium demonstrated that the ED pathway is functional in V. cholerae and is obligatory for gluconate catabolism. Importantly, selective activation of the ED pathway led to concurrent elevation of transcripts of prime virulence genes (ctxA and tcpA) and their regulator (toxT). Further, lowering of these transcript levels and cholera toxin production in vitro by an ED pathway-defective mutant (strain N16961 with a Δedd mutation [Δedd(N16961) strain]) suggested the importance of this pathway in regulating V. cholerae virulence. The in vivo relevance of these data was established as the mutant failed to colonize in suckling mice intestine or to induce fluid accumulation in ligated rabbit ileal loops. Activation of the ED pathway in V. cholerae was shown to inhibit biofilm formation in vitro that could be reversed in the mutant. As further support for these results, comparative transcriptome analysis with cells grown in the presence of glucose or gluconate revealed that a functional ED pathway led to activation of a subset of previously reported in vivo expressed genes. All of these results suggest the importance of the ED pathway in V. cholerae pathogenesis.
Asunto(s)
Aldehído-Liasas/metabolismo , Cólera/microbiología , Regulación Bacteriana de la Expresión Génica , Gluconatos/metabolismo , Hidroliasas/metabolismo , Vibrio cholerae/patogenicidad , Aldehído-Liasas/genética , Animales , Animales Lactantes , Medios de Cultivo , Modelos Animales de Enfermedad , Perfilación de la Expresión Génica , Silenciador del Gen , Hidroliasas/genética , Intestinos/microbiología , Ratones , Conejos , Vibrio cholerae/genética , Vibrio cholerae/crecimiento & desarrollo , Vibrio cholerae/metabolismo , Virulencia , Factores de Virulencia/genética , Factores de Virulencia/metabolismoRESUMEN
BACKGROUND & OBJECTIVES: Cyclone AILA hit Indian States on eastern coast on May 25, 2009. An investigation was conducted to examine if AILA was responsible for increased reporting of diarrhoea cases from the district of East-Medinipur in West Bengal. Identifying causative organisms for diarrhoea and assessing their antibiotic susceptibility profile were other objectives. METHODS: Rapid situation and response assessment technique was employed to triangulate primary and secondary data collected through field visits. Prescription audit was also conducted. RESULTS: Significantly increased occurrence of diarrhoea was observed in June 2009 in two subdivisions namely Haldia and Egra (OR 1.6 and 1.3 respectively; 95% CI 1.52-1.65 and 1.21-1.32 P<0.001) considering 2007 as baseline. Vibrio cholerae grew from 54 per cent of the stool samples (21/39; 17 V. cholerae O1-Ogawa and 4 non-O1-non-O139), confirming a community outbreak of cholera. Shigella flexneri 3a was isolated from 5 per cent stool specimens. Increased rate of admission in treatment centres due to diarrhoea in the whole district coincided with the formation of cyclone and showed over two-fold rise compared to the admission recorded 6 days ago. Haldia subdivision had the highest attack rate of 9 per 1000 in the month of June, 2009 whereas for the whole district it was 5 per 1000 in the same month. All the isolates of V. cholerae were resistant to ampicillin and furazolidone and sensitive to norfloxacin and azithromycin. INTERPRETATION & CONCLUSIONS: Pre-AILA changes in the environment, AILA and seasonality of diarrhoea in the study district interplayed towards increased occurrence of diarrhoea. Continuous tracking of 'seasonality of diarrhoea in the community with vulnerability assessment of potential hosts', 'antibiotic sensitivity profile of the causative microorganisms', and 'prescription practice of physicians' would help appropriate disaster management.
Asunto(s)
Cólera/epidemiología , Tormentas Ciclónicas , Diarrea/epidemiología , Brotes de Enfermedades/estadística & datos numéricos , Medicina Ambiental , Vibrio cholerae/aislamiento & purificación , Vibrio cholerae/patogenicidad , Antibacterianos , Estudios Transversales , Desastres , Retroalimentación , Humanos , India/epidemiología , Salud PúblicaRESUMEN
The rise in multi-drug resistant Vibrio cholerae strains is a big problem in treatment of patients suffering from severe cholera. Only a few studies have evaluated the potential of natural compounds against V. cholerae. Extracts from plants like 'neem', 'guazuma', 'daio', apple, hop, green tea and elephant garlic have been shown to inhibit bacterial growth or the secreted cholera toxin (CT). However, inhibiting bacterial growth like common antimicrobial agents may also impose selective pressure facilitating development of resistant strains. A natural compound that can inhibit virulence in V. cholerae is an alternative choice for remedy. Recently, some common spices were examined to check their inhibitory capacity against virulence expression of V. cholerae. Among them methanol extracts of red chili, sweet fennel and white pepper could substantially inhibit CT production. Fractionation of red chili methanol extracts indicated a hydrophobic nature of the inhibitory compound(s), and the n-hexane and 90 per cent methanol fractions could inhibit >90 per cent of CT production. Purification and further fractionation revealed that capsaicin is one of the major components among these red chili fractions. Indeed, capsaicin inhibited the production of CT in various V. cholerae strains regardless of serogroups and biotypes. The quantitative reverse transcription real-time PCR assay revealed that capsaicin dramatically reduced the expression of major virulence-related genes such as ctxA, tcpA and toxT but enhanced the expression of hns gene that transcribes a global prokaryotic gene regulator (H-NS). This indicates that the repression of CT production by capsaicin or red chili might be due to the repression of virulence genes transcription by H-NS. Regular intake of spices like red chili might be a good approach to fight against devastating cholera.
Asunto(s)
Antibacterianos/farmacología , Productos Biológicos , Farmacorresistencia Bacteriana , Extractos Vegetales/farmacología , Vibrio cholerae/efectos de los fármacos , Vibrio cholerae/patogenicidad , Animales , Antibacterianos/uso terapéutico , Capsaicina/farmacología , Capsaicina/uso terapéutico , Cólera/tratamiento farmacológico , Contraindicaciones , Diarrea/tratamiento farmacológico , Humanos , Extractos Vegetales/uso terapéuticoRESUMEN
The Gram-negative bacteria Vibrio cholerae poses significant public health concerns by causing an acute intestinal infection afflicting millions of people each year. V. cholerae motility, as well as virulence factor expression and outer membrane protein production, has been shown to be affected by bile. The current study examines the effects of bile on V. cholerae phospholipids. Bile exposure caused significant alterations to the phospholipid profile of V. cholerae but not of other enteric pathogens. These changes consisted of a quantitative increase and migratory difference in cardiolipin, decreases in phosphatidylglycerol and phosphatidylethanolamine, and the dramatic appearance of an unknown phospholipid determined to be lyso-phosphatidylethanolamine. Major components of bile were not responsible for the observed changes, but long-chain polyunsaturated fatty acids, which are minor components of bile, were shown to be incorporated into phospholipids of V. cholerae. Although the bile-induced phospholipid profile was independent of the V. cholerae virulence cascade, we identified another relevant environment in which V. cholerae assimilates unique fatty acids into its membrane phospholipids - marine sediment. Our results suggest that Vibrio species possess unique machinery conferring the ability to take up a wider range of exogenous fatty acids than other enteric bacteria.
Asunto(s)
Membrana Celular/metabolismo , Ácidos Grasos/metabolismo , Interacciones Huésped-Patógeno , Vibrio cholerae/citología , Microbiología del Agua , Bilis/metabolismo , Ácidos y Sales Biliares/metabolismo , Bilirrubina/metabolismo , Escherichia coli/metabolismo , Ácidos Grasos Insaturados/metabolismo , Sedimentos Geológicos/química , Lecitinas/metabolismo , Fosfolípidos/química , Fosfolípidos/aislamiento & purificación , Salmonella enterica/metabolismo , Especificidad de la Especie , Espectrometría de Masa por Ionización de Electrospray , Vibrio cholerae/crecimiento & desarrollo , Vibrio cholerae/metabolismo , Vibrio cholerae/patogenicidad , VirulenciaRESUMEN
Carvacrol and cymene, phenolic compounds naturally present in the essential oil of oregano and thyme, were examined for their antimicrobial activity against Vibrio cholerae (ATCC 14033, VC1, and VC7) inoculated in carrot juice. Carvacrol exhibited a dose dependent inhibitory effect on the bacteria. Although cymene did not have antimicrobial activity against the bacteria, it enhanced the inhibitory ability of carvacrol. At 25 °C, the lowest concentrations of carvacrol and cymene required for zero detectable viable count varied depending on bacterial strains; 5 and 5 ppm, respectively, for VC7; 5 and 7.5 ppm, respectively, for VC1; and 7.5 and 7.5 ppm, respectively, for ATCC 14033. This study also examined several factors influencing the antimicrobial activity of carvacrol and cymene against V. cholerae ATCC 14033, including temperature, bacterial cell number, and food substrate. Carvacrol and cymene inhibited the bacterium in carrot juice at 25 °C more efficiently than at 15 and 4 °C. The doses of both compounds required for zero detectable viable count increased as the number of the bacterial cells in the carrot juice increased. The fat content and the complexity of foods were shown to decrease the antimicrobial activity of the compounds.
Asunto(s)
Antiinfecciosos/farmacología , Microbiología de Alimentos , Monoterpenos/farmacología , Vibrio cholerae/efectos de los fármacos , Vibrio cholerae/patogenicidad , Cólera/prevención & control , Cimenos , Conservación de Alimentos , Humanos , Aceites Volátiles/química , Origanum/química , Aceites de Plantas/química , Thymus (Planta)/química , Vibrio cholerae/crecimiento & desarrolloAsunto(s)
Antibacterianos/farmacología , Regulación Bacteriana de la Expresión Génica/fisiología , Myrica , Vibrio cholerae/crecimiento & desarrollo , Vibrio cholerae/genética , Animales , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Humanos , Ratones , Extractos Vegetales/farmacología , Vibrio cholerae/efectos de los fármacos , Vibrio cholerae/patogenicidad , VirulenciaRESUMEN
Silkworm larvae, Bombyx mori, were examined as an animal model of human infection with pathogenic bacteria. When 3 x 10(7) cells of Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa, or Vibrio cholerae were injected into the blood of fifth instar silkworm larvae, over 90% of the larvae died within 2 days, whereas over 90% survived for 5 days after injection of the same amount of Escherichia coli. Growth of S. aureus was observed in larvae blood and tissues. Immunostaining analysis revealed that S. aureus proliferated at the surface of the midgut. Infection of silkworm larvae by methicillin-sensitive S. aureus was cured by ampicillin, oxacillin, and vancomycin, whereas infection by methicillin-resistant S. aureus was not cured by ampicillin or oxacillin, although vancomycin was effective. Disinfectants were not effective because of toxicity against the larvae. Thus, silkworm larvae are useful for evaluating antibiotics for pathogenic bacterial infection in humans.
Asunto(s)
Antibacterianos/uso terapéutico , Infecciones Bacterianas/tratamiento farmacológico , Bombyx/microbiología , Larva/microbiología , Pseudomonas aeruginosa/fisiología , Staphylococcus aureus/fisiología , Vibrio cholerae/fisiología , Animales , Antibacterianos/farmacología , Infecciones Bacterianas/microbiología , Bombyx/efectos de los fármacos , Bombyx/crecimiento & desarrollo , Modelos Animales de Enfermedad , Quimioterapia Combinada/farmacología , Quimioterapia Combinada/uso terapéutico , Humanos , Concentración 50 Inhibidora , Larva/efectos de los fármacos , Larva/fisiología , Dosificación Letal Mediana , Resistencia a la Meticilina , Pruebas de Sensibilidad Microbiana , Oxazoles/farmacología , Oxazoles/uso terapéutico , Pseudomonas aeruginosa/crecimiento & desarrollo , Pseudomonas aeruginosa/patogenicidad , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , Staphylococcus aureus/patogenicidad , Vancomicina/farmacología , Vancomicina/uso terapéutico , Vibrio cholerae/crecimiento & desarrollo , Vibrio cholerae/patogenicidadRESUMEN
El vibrio cholerae tipo no-01 es un microorganismo de distribución mundial con hábitat acuático, que ocasionalmente produce patología en el hombre. Existe relación directa demostrada entre la ingesta de productos de mar y/o la realización de actividades marinas, y la infección por vibrio. La clínica que produce con más frecuencia es la gastrointestinal y en particular la enfermedad diarreica. Por otro lado, la otitis media por vibrio cholerae tipo no01 es extremadamente infrecuente en nuestro medio, aunque si se revisa la bibliografía, su incidencia parece haber aumentado durante los últimos años. Se presenta un caso clínico de otitis media supurada por vibrio cholerae tipo no-01 y se revisa la literatura. (AU)
Asunto(s)
Masculino , Niño , Humanos , Vibrio cholerae/aislamiento & purificación , Vibrio cholerae/patogenicidad , Amoxicilina/administración & dosificación , Amoxicilina/uso terapéutico , Penicilinas/aislamiento & purificación , Penicilinas/análisis , Quinolonas/aislamiento & purificación , Quinolonas/análisis , Otitis Media Supurativa/diagnóstico , Otitis Media Supurativa/etiología , Otitis Media Supurativa/tratamiento farmacológico , Ciprofloxacina/uso terapéutico , Pruebas de Sensibilidad Microbiana/métodos , Antibacterianos/administración & dosificación , Antibacterianos/uso terapéutico , Vibrio cholerae/inmunología , Vibrio cholerae/ultraestructura , Dolor de Oído/diagnóstico , Dolor de Oído/etiología , Hepatopatías/diagnóstico , Hemocromatosis/diagnóstico , Hemocromatosis/complicaciones , Otitis Media Supurativa/epidemiología , Otitis Media Supurativa/microbiologíaRESUMEN
Con el fin de estudiar los patrones de excreción, colonización y la capacidad protectora de cepas vivas atenuadas de Vibrio cholerae O1El Tor, se inmunizaron conejos Nueva Zelandia con estas cepas y su correspondiente parental, con 2 dosis por el modelo de inoculación oral en conejos adultos. Fueron retados 2 semanas después de la segunda dosis por el modelo de intestino ligado, con cepas altamente virulentas de V. choleraeO1 serotipos Ogawa e Inaba y serogrupo O139. Se comprobó que las cepas manupuladas de forma genética no afectan los patrones de excreción, cuando se compara con su parental. Se observó en el reto una disminución en los niveles de colonización de las cepas virulentas de ambos serotipos; tanto en los conejos inmunizados con las cepas atenuadas como con la parental en comparación con animales controles inmunizados con la cepa Escherichia coli K-12, lo que indica que hubo cierto grado de protección. En el caso de los animales retados con la cepa 0139 se demostró que la protección es específica para cada serogrupo pues en este caso no se observó disminución de la colonización.
In order to study the excretion patterns, colonization and protective capacity of live sttenuated strains of Vibrio cholerae O1. E1 Tor, rabbits were immunized in New Zealand with these strains and their corresponding parental strains. 2 doses were administered by the model of oral inoculation in adult rabbits. Rabbits were rotated 2 weeks after the second dose by the model of ligated intestine with highly virulent strains of V. cholerae O1 Ogawa and Inaba serotypes and O139 serogroup. It was proved that the genetically manipulated strains do not effect the excretion patterns when they are compared with their parental strains. It was observed in the challenge a decrease in the levels of colonization of virulent strains of both serotypes, not only among the rabbits immunized with the attenuated strains, but also among those immunizedwith the parental strains in comparison with control animals immunized with the strain of Escherichia coli K-12, which means that there was certain degree of protection. In the case of the animals challenged with the O139 strain it was demonstrated that the protection is specific for each serogroup, since in this case there was no reduction of the colonization.