Corchorus olitorius aqueous extract attenuates quorum sensing-regulated virulence factor production and biofilm formation

Objective:To investigate the effect of Corchorus olitorius aqueous fraction (COAF) on quorum sensing (QS)-regulated virulence factors and biofilm formation in Pseudomonas aeruginosa (PAO1). Methods:The preliminary screening of the anti-QS effect of COAF was performed by evaluating the anti-pathogenic activity against Chromobacterium violaceum CV026 biosensor strain. Next, the inhibitory effects of COAF on QS-regulated pyocyanin production, proteolytic and elastolytic activities, swarming motility, and biofilm formation were evaluated in PAO1.Results:The results showed that the treatment of COAF significantly decreased the biofilm biomass, attenuated virulence factors, and inhibited swarming motility of PAO1 without affecting the growth of the bacteria in a dose-dependent manner. COAF at 2000 μg/mL significantly decreased Las B elastase activity in PAO1 culture, exopolysaccharide production, swarming motility, pyocyanin level, and biomass of PAO1 by 55％ (P<0.05), 60％ (P<0.01), 61％ (P<0.01), 65％(P<0.01) and 73％ (P<0.01), respectively. In addition, the production of violacein was decreased by 62％ (P<0.01) with the treatment of a high dose of COAF. Conclusions:These findings indicate that COAF can be a potential source of anti-QS agents.

Septicemia por Chromobacterium Violaceum en pediatría / Septicemia due to Chromobacterium Violaceum in pediatrics

Chromobacterium violaceum es una bacteria gram negativa anaerobia facultativa, que se encuentra ampliamente distribuida en el agua y el suelo en regiones tropicales y subtropicales, que se asocia con infecciones respiratorias, gastrointestinales, abscesos hepáticos, meningitis, endocarditis, síndrome hemofagocítico y sepsis fulminante. Se presentan 2 casos en niños: el primero es un varón de 8 años con lesiones en piel, fiebre y adenitis inguinal, que ingresó con un cuadro de sepsis severa, síndrome de distrés respiratorio agudo (SDRA) y falleció a las 3 h del ingreso. De los hemocultivos se aisló Chromobacterium violaceum. El segundo caso, es una niña de 12 años con antecedente de fiebre y adenopatía inguinal secundaria a herida cortopunzante en el pie homolateral, que ingresó con un cuadro de sepsis, con desarrollo de abscesos múltiples profundos. De la colección obtenida de piel y partes blandas y de un aspirado traqueal se aisló Chromobacterium violaceum. Recibió tratamiento antibiótico adecuado y posteriormente fue dada de alta. Se realizó una revisión bibliográfica de esta infección en niños y se encontraron 44 casos en todo el mundo. Algunos de éstos, se relacionaron con inmunodeficiencia de base, como la enfermedad granulomatosa crónica. La infección por esta bacteria es rara y se presenta como un cuadro grave que no responde a antibióticos habituales de uso empírico y tiene una alta tasa de mortalidad (AU)

Chromobacterium violaceum is a facultative anaerobic Gramnegative bacillus, widely distributed in water and soil in tropical and subtropical regions and associated with respiratory and gastrointestinal infections, liver abscesses, meningitis, endocarditis, hemophagocytic syndrome, and fulminant sepsis. Here two pediatric cases are presented: The first was an 8-year-old boy with skin lesions, fever, and inguinal adenitis, who was admitted with severe sepsis, acute respiratory distress syndrome (ARDS) and died three hours after. Chromobacterium violaceum was isolated from blood cultures. The second case was a 12-year-old girl with a history of fever and inguinal adenopathy secondary to a wound in the homolateral foot, who was admitted because of sepsis and multiple deep abscesses. From samples collected from the skin and soft tissues as well as tracheal aspirate Chromobacterium violaceum was isolated. Adequate antibiotic treatment was started and the patient was subsequently discharged. In a review of the literature, 44 cases worldwide were identified. Some of these cases were related to underlying immunodeficiency, such as chronic granulomatous disease. Infection with this bacterium is rare and presents with severe manifestations that do not respond to the common empirical antibiotics and are associated with a high mortality rate (AU)

Anti-Candida and anti-quorum sensing activity of airborne microorganisms detected by a rapid method

Abstract INTRODUCTION: Introducing new antibiotics to the clinic is critical. METHODS: We adapted a plate method described by Kawaguchi and coworkers in 20131 for detecting inhibitory airborne microorganisms. RESULTS: We obtained 51 microbial colonies antagonist to Chromobacterium violaceum, purified and retested them, and of these, 39 (76.5%) were confirmed. They comprised 24 bacteria, 13 fungi, and 2 yeasts. Among the fungi, eight (61.5%) produced active extracts. Among the bacterial, yeast, and fungal strains, 17 (44.7%) and 12 (31.6%) were active against Candida albicans and Candida parapsilosis, respectively. CONCLUSIONS: The proposed screening method is a rapid strategy for discovering potential antibiotic producers.

Romipeptides A and B, two new romidepsin derivatives isolated from Chromobacterium violaceum No.968 and their antitumor activities in vitro / 中国天然药物

Romipeptides A and B (1 and 2), two new romidepsin derivatives, and three known compounds, chromopeptide A (3), romidepsin (4) and valine-leucine dipeptide (5) were isolated from the fermentation broth of Chromobacterium violaceum No. 968. Their structures were elucidated by interpretation of their UV, HR-ESI-MS and NMR spectra. The absolute configuration of compound 1 and 2 were established by single crystal X-ray diffraction analysis. Compounds 1-5 were evaluated for their anti-proliferative activities against three human cancer cell lines, SW620, HL60, and A549. The results showed most of these compounds exhibited antitumor activities in vitro, in which compound 2 displayed potent cytotoxicity to SW620, HL60 and A549 cell lines, with IC of 12.5, 6.7 and 5.7 nmol·L, respectively.

Reporte de dos casos de inusual infección no letal por Chromobacterium violaceum. Revisión literaria / Report two cases non lethal infectious diseases for Chromobacterium violaceum. Review of the literature

Chromobacterium violaceum es una bacteria gramnegativa anaeróbica facultativa, que habita en el suelo y el agua de las áreas tropicales y subtropicales. La infección en seres humanos es rara. A continuación se presentan dos casos; una lactante mayor, indígena, quien posterior a baño de inmersión en aguas estancadas comienza a presentar fiebre con aumento de volumen y limitación funcional y gonalgia izquierda, se realiza lavado articular, se toma muestra para cultivo con crecimiento para Chromobacterium violaceum. Un escolar masculino de 7 años de edad con antecedente de Leucemia Linfoblastica Aguda con recaída en sistema nervioso central, cumpliendo esquema de altas dosis de metotrexate; quien comienza a presentar fiebre y signos de flogosis en sitio de colocación de cáteter de vía central, el cual es retirado y cultivado con crecimiento para Chromobacterium violaceum. Esta infección en humanos es una enfermedad grave y sistémica con una alta tasa de mortalidad.

Chromobacterium violaceum is a Gram-negative facultative anaerobic bacterium, which lives in the soil and water of subtropical and tropical areas. Infection in humans is rare. Here we present two cases. One was in one indigenous newborn, who after one immersion in stagnant waters presented fever and increase in the volume of testicle and limitation in movility of the left limb. After arthrocentesis of left knee and culture of the sinovial liquid, growth of Cromabacterium violaceum was reported. Another case was a male school of 7 years old with a history of acute lymphoblastic leukemia relapse in the central nervous system, that completed his high dose methotexate scheme. Some days after he presented fever and edema at the site of central venous catheterization. The catheter was retired and it was obtained growth of Chromobacterium violaceum. Human infection with Chromobacterium violaceum is a severe and systemic disease with a high mortality rate.

Recent research advances on Chromobacterium violaceum

Chromobacterium violaceum is a gram-negative bacterium, which has been used widely in microbiology labs involved in quorum sensing (QS) research. Among the QS-regulated traits of this bacterium, violacein production has received the maximum attention. Violacein production in this organism, however is not under sole control of QS machinery, and other QS-regulated traits of this bacterium also need to be investigated in better detail. Though not often involved in human infections, this bacterium is being viewed as an emerging pathogen. This review attempts to highlight the recent research advances on C. violaceum, with respect to violacein biosynthesis, development of various applications of this bacterium and its bioactive metabolite violacein, and its pathogenicity.

Recent research advances on Chromobacterium violaceum

Chromobacterium violaceum is a gram-negative bacterium, which has been used widely in microbiology labs involved in quorum sensing (QS) research. Among the QS-regulated traits of this bacterium, violacein production has received the maximum attention. Violacein production in this organism, however is not under sole control of QS machinery, and other QS-regulated traits of this bacterium also need to be investigated in better detail. Though not often involved in human infections, this bacterium is being viewed as an emerging pathogen. This review attempts to highlight the recent research advances on C. violaceum, with respect to violacein biosynthesis, development of various applications of this bacterium and its bioactive metabolite violacein, and its pathogenicity.

Composition, anti-quorum sensing and antimicrobial activity of essential oils from Lippia alba

Many Gram-negative pathogens have the ability to produce N-acylhomoserine lactones (AHLs) as signal molecules for quorum sensing (QS). This cell-cell communication system allows them to coordinate gene expression and regulate virulence. Strategies to inhibit QS are promising for the control of infectious diseases or antibiotic resistant bacterial pathogens. The aim of the present study was to evaluate the anti-quorum sensing (anti-QS) and antibacterial potential of five essential oils isolated from Lippia alba on the Tn-5 mutant of Chromobacterium violaceum CV026, and on the growth of the gram-positive bacteria S. aureus ATCC 25923. The anti-QS activity was detected through the inhibition of the QS-controlled violacein pigment production by the sensor bacteria. Results showed that two essential oils from L. alba, one containing the greatest geranial:neral and the other the highest limonene:carvone concentrations, were the most effective QS inhibitors. Both oils also had small effects on cell growth. Moreover, the geranial/neral chemotype oil also produced the maximum zone of growth inhibition against S. aureus ATCC 25923. These data suggest essential oils from L. alba have promising properties as QS modulators, and present antibacterial activity on S. aureus.

Chromobacterium violaceum: um caso fatal no nordeste do Brasil / Chromobacterium violaceum: a fatal case in the northeast of the Brazil

Chromobacterium violaceum is a rare pathogen that can potentially cause fatal infections in humans. An 8-year-old child from Natal, northeast of Brazil, presented history of fever, sore throat, and abdominal pain, during 5 days before admission, and died 4 hours after hospitalization. Chromobacterium violaceum was isolated from oropharynx scrapings and was resistant to ampicillin, cefotaxime, cefalotin, ceftazidime, and ceftriaxone...

Chromobacterium violaceum é um patógeno raro que potencialmente pode causar infecções fatais em humanos. Relatamos o caso de uma criança de 8 anos de idade, moradora da cidade de Natal, nordeste do Brasil, que apresentou história de febre, dor na garganta e no abdome durante os cinco dias anteriores à internação, e veio a falecer após 4 horas de hospitalização. A bactéria Chromobacterium violaceum foi isolada da orofaringe e apresentou resistência a ampicilina, cefotaxima, cefalotina, ceftazidima e ceftriaxona...

Biosurfactant production by Chr omobacterium violaceum ATCC 12472 using corn steep liquor and corn post-frying oil as nutrients / Produção de biossurfactante por Chromobacterium violaceum ATCC 12472 utilizando milhocina e óleo de milho pós-fritura como nutrientes

The interest in microbial surfactants has been increased considerably in the recent years. Environmental concern among consumers, combined with new laws control of the environment, the demand for natural surfactants, to replace petrochemicals. The microbial surfactants have been tested in many industrial and environmental applications, such as in bioremediation, oil dispersion and oil recovery, replacing chemical surfactants. Furthermore, they can also be used in the food, cosmetic, detergent and agriculture. However, increased environmental concern among consumers to new legislation combined control of the environment has led to demand for natural surfactants as alternatives to existing products. This work aimed to produce biosurfactant by Chromobacterium violaceum ATCC 12472, using low nutrient value. The surface tension of the biosurfactant produced by C. violaceum ATCC 12472 had values between 29 and 40 mN/m. The best result was observed on the assay 8 (8.0% of corn steep liquor, 1.0% of lactose and corn oil post-frying 7.5%) factorial design, within 72 hours of fermentation, lowering the surface tension of water from 71 to 29 mN/m. The best results of the index (E24) and emulsification activity (UEA) were observed in the central point, and using the both methods and corn post-frying oil resulted in 72% of emulsification, and the emulsification activity value of 6 UAE. The results demonstrate the high potential of C. violaceum promising in producing a biosurfactant.

O interesse em surfactantes de origem microbiana tem aumentado consideravelmente nos últimos anos. A preocupação ambiental entre os consumidores, combinada às novas legislações de controle do meio ambiente, aumentou a procura por surfactantes naturais em substituição aos derivados petroquímicos. Os surfactantes microbianos têm sido testados em muitas aplicações ambientais e industriais, como na biorremediação, na dispersão de manchas oleosas e na recuperação de petróleo, substituindo os surfactantes químicos. Além disso, também podem ser utilizados nas indústrias de alimentos, cosméticos, detergentes e na agricultura. Neste trabalho foi produzido biossurfactante por Chromobacterium violaceum ATCC 12472, utilizando nutrientes de baixo valor agregado. A tensão superficial do biossurfactante produzido por C. violaceum ATCC 12472 teve valores entre 29 e 40 mN/m. O melhor resultado foi observado no ensaio 8 (milhocina 8,0%, lactose 1,0% e óleo de milho pós-fritura 7,5%) do planejamento fatorial, com 72 horas de fermentação, reduzindo a tensão superficial da água de 71 para 29 mN/m. Os melhores resultados do índice (E24) e da atividade de emulsificação (UEA) foram observados no ponto central, sendo utilizados para ambos os métodos o óleo de milho, que resultou no E24 de 2% e atividade com valores acima de 6 UAE. Os resultados obtidos demonstram o elevado potencial de C. violaceum na produção de um promissor biossurfactante.

Antipathogenic potential of marine Bacillus sp. SS4 on N-acyl-homoserine-lactone-mediated virulence factors production in Pseudomonas aeruginosa (PAO1).

Antipathogenic therapy is an outcome of the quorum-sensing inhibition (QSI) mechanism, which targets autoinducerdependent virulent gene expression in bacterial pathogens. N-acyl homoserine lactone (AHL) acts as a key regulator in the production of virulence factors and biofilm formation in Pseudomonas aeruginosa PAO1 and violacein pigment production in Chromobacterium violaceum. In the present study, the marine bacterial strain SS4 showed potential QSI activity in a concentration-dependent manner (0.5–2 mg/ml) against the AHL-mediated violacein production in C. violaceum (33–86%) and biofilm formation (33–88%), total protease (20–65%), LasA protease (59– 68%), LasB elastase (36–68%), pyocyanin (17–86%) and pyoverdin productions in PAO1. The light and confocal laser scanning microscopic analyses confirmed the reduction of the biofilm-forming ability of PAO1 when treated with SS4 extract. Furthermore, the antibiofilm potential was confirmed through static biofilm ring assay, in which ethyl acetate extract of SS4 showed concentration-dependent reduction in the biofilm-forming ability of PAO1. Thus, the result of this study clearly reveals the antipathogenic and antibiofilm properties of the bacterial isolate SS4. Through 16S rDNA analysis, the strain SS4 was identified as Bacillus sp. (GenBank Accession Number: GU471751).

Screening of certain medicinal plants from India for their anti-quorum sensing activity.

Discovery of quorum sensing (QS) system to coordinate virulence and biofilm formation in bacterial pathogens has triggered search for safe, stable and non-toxic anti-QS compounds from natural products. Ethanolic extracts of 24 Indian medicinal plants were tested by agar well and disc diffusion assay for anti-QS activity using Chromobacterium violaceum (CV12472 and CVO26) reporter strains. AHL from C. violaceum CV31532 was isolated and partially purified for its use in CVO26 based bioassay. Effect on swarming-motility of Pseudomonas aeruginosa (PAO1) was also recorded at sub-MIC concentrations of extracts. Of the 24 medicinal plants screened Hemidesmus indicus (L.) Schult (root), Holarrhena antidysenterica (Roth)A.DC. (bark), Mangifera indica L. (seed) Punica granatum L. (pericarp) and Psoralea corylifolia L. (seed) demonstrated varying level of inhibition of violacein production in the reporter strains. Moreover, a significant reduction in swarms was recorded over control. The inhibition of violacein production and swarming motility may be due to direct or indirect interference on QS by active constituents or the interactive effect of different phytocompounds present in the extracts. These plant extracts may be selected for activity guided fractionation to identify and characterize the active principle

Computational analysis suggests that virulence of Chromobacterium violaceum might be linked to biofilm formation and poly-NAG biosynthesis

Groups of genes that produce exopolysaccharide with a N-acetyl-D-glucosamine monomer are in the genome of several pathogenic bacteria. Chromobacterium violaceum, an opportunistic pathogen, has the operon hmsHFR-CV2940, whose proteins can synthesize such polysaccharide. In this work, multiple alignments among proteins from bacteria that synthesize such polysaccharide were used to verify the existence of amino acids that might be critical for pathogen activity. Three-dimensional models were generated for spatial visualization of these amino acid residues. The analysis carried out showed that the protein HmsR preserves the amino acids D135, D228, Q264 and R267, considered critical for the formation of biofilms and, furthermore, that these amino acids are close to each other. The protein HmsF of C. violaceum preserves the residues D86, D87, H156 and W115. It was also shown that these residues are also close to each other in their spatial arrangement. For the proteins HmsH and CV2940 there is evidence of conservation of the residues R104 and W94, respectively. Conservation and favorable spatial location of those critical amino acids that constitute the proteins of the operon indicates that they preserve the same enzymatic function in biofilm synthesis. This is an indicator that the operon hmsHFR-CV2940 is a possible target in C. violaceum pathogenicity.

Diversity of Chromobacterium violaceum isolates from aquatic environments of state of Pará, Brazilian Amazon

The present study intended to characterize the phenotypic and genetic diversity of Brazilian isolates of Chromobacterium violaceum from aquatic environments within the Amazon region. Nineteen isolates showed morphological properties of C. violaceum and the majority grew at 44°C. Low temperatures, in contrast, showed to be inhibitory to their growth, as eleven isolates did not grow at 10ºC and nine did not produce pigmentation, clearly indicating an inhibition of their metabolism. The largest variation among isolates was observed in the citrate test (Simmons), in which 12 isolates were positive, and in the oxidation/fermentation of sucrose, with six positives isolates. Chloramphenicol, gentamicin and sulfonamides efficiently inhibited bacterial growth. Amplified products of the recA gene were digested with HindII or PstI, which produced three or four restriction fragments patterns, respectively. The combined analysis arranged the isolates into six genospecies. The higher diversity observed in Belém (genotypes C, D, E and F) may be a consequence of intense human occupation, pollution of the aquatic environment or due to the higher diversity of the environments sampled in that region. In conclusion, a high level of genetic and phenotypic diversity was observed, and four new genospecies were described.

Isolation and Identification of Marine Actinomycetes with Quorum Sensing Inhibitory Activity / 微生物学通报

Quorum sensing systems of pathogens are central regulators for the expression of virulence factors. Increasing evidence implies that targeting the quorum sensing system of many pathogenic bacteria is a promising therapeutic approach to control infections. In this work,we isolated 47 strains of actinomycetes from the mud sample of Jiaozhou Bay. Quorum sensing inhibitory activity was monitored by Chromobacterium violaceum CV026. As a result,the culture broth extract of actinomycetes WA-7 was found to have significant quorum sensing inhibitory activity. This strain was assigned to the genus Streptomyces based on its 16S rDNA sequence. Further investigation revealed that the extract could inhibit the quorum sensing-controlled violacein and proteases production of C. violaceum in a concentration-dependent manner.

Fatal septicemia caused by Chromobacterium violaceum in a child from Colombia

A 4-year old child living in Colombia presented with a history of fever and severe abdominal pain for four days. The patient developed pneumonia, septic shock, multiple organ failure and died on the fifth day of hospitalization. Chromobacterium violaceum was isolated from admission blood cultures and was resistant to ampicillin, cephalosporins, carbapenems and aminoglycosides.

Una niña de 4 años que vivía en Colombia presentó historia de fiebre y dolor abdominal severo por cuatro días. La paciente desarrollo neumonía, shock séptico, múltiple falla de órganos y muerte el quinto día de hospitalización. Chromobacterium violaceum fue aislado de cultivos de sangre y mostró resistencia a ampicilina, cefalosporinas, carbapenems y aminoglicosidos.

Sepsis por Chromobacterium violaceum pigmentado y no pigmentado / Sepsis caused by pigmented and no pigmented Chromobacterium violaceum

La sepsis por Chromobacterium violaceum es un cuadro clínico rara vez reportado y asociado con una elevada letalidad. Comunicamos un caso fatal de sepsis en un niño indígena venezolano con 6 años de edad, quien presentó fiebre y aumento de volumen en la región inguinal derecha y que fuera inicialmente diagnosticado como plastrón apendicular abscedado. La cirugía de apendicetomía reveló un absceso en el músculo psoas derecho, del cual se aisló una cepa pigmentada de C. violaceum, mientras que a partir de hemocultivos se aislaron cepas pigmentada y no pigmentada, simultáneamente, del mismo agente. Se administró tratamiento con imipenem y amikacina pero su condición se deterioró y falleció a los 9 días de su ingreso.

Chromobacterium violaceum sepsis is rare but associated with a high mortality rate. We report a fatal case of C. violaceum sepsis in a 6 years old Venezuelan indian boy. Clinical manifestations were fever and swelling in the right inguinal region. The initial diagnosis was an appendicular plastron. Appendicectomy was performed and during surgery a right psoas abscess was identified that resulted culture positive for pigmented C. violaceum. Blood cultures were positive for a pigmented and non pigmented C. violaceum strain. Imipenem and amikacin were administered despite of which the child died 9 days after hospital admission.

Sepsis by Chromobacterium violaceum: first case report from Colombia

Chromobacterium violaceum is found in tropical and subtropical regions; it is the only Chromobacterium species pathogenic for humans. Due to its rare presentation, physicians often ignore the importance of this pathogen. We report a fulminant fatal case of bacteremia in a 38-year-old Colombian man. The clinical manifestations were fever, thoracic pain, respiratory failure and death. His condition, from the beginning of clinical diagnosis, went into continuous deterioration, till his death, within a few days after the symptoms began. Two hemocultures isolated C. violaceum. We conclude that doctors should consider this differential diagnosis in patients with systemic inflammatory response syndrome, with continuous deterioration.

A Case of Chromobacterium Infection after Car Accident in Korea

Chromobacterium violaceum is a gram negative straight rod, 0.8-1.2 by 2.5 to 6.0 m, which is motile by one polar flagella and one to four lateral flagella. The organism inhabits soil and water and is often found in semitropical and tropical climates. Infections in humans are rare. We report a case of infection caused by strains of C. violaceum. A 38-year-old male patient was admitted to KyungHee University Hospital, Seoul, Korea on July 28th, 2003, after a car accident. The patient had multiple trauma and lacerations. He had an open wound in the left tibial area from which C. violaceum was isolated. The strain was resistant to ampicillin, tobramycin, ampicillin/sulbactam, ceftriaxone and cefepime, but was susceptible to amikacin, gentamicin, ciprofloxacin, levofloxacin, trimethoprim/sulfamethoxazole and piperacillin/tazobactam. The patient was treated successfully by debridement, cephapirin sodium and astromicine sulfate.