Evaluation of anti-quorum sensing potential of Averrhoa bilimbi (Kamias) against Pseudomonas aeruginosa ATCC 27853

BACKGROUND AND OBJECTIVE@#Many opportunistic and nosocomial pathogens like Pseudomonas aeruginosa are very reliant on a bacterium-to-bacterium communication system called quorum sensing (QS). Without the aforementioned process, gene expressions associated with virulence factors will not be produced. In this study, the sub-inhibitory concentrations (sub-MICs) of methanolic leaf extract and obtained fractions from Averrhoa bilimbi (kamias) were screened for ability to inhibit quorum sensing-controlled phenotypes of P. aeruginosa ATCC 27853.@*METHODOLOGY@#A. bilimbi crude extract was fractionated through liquid-liquid extraction, producing four (4) fractions: hexane fraction, dichloromethane (DCM) fraction, ethyl acetate (EtOAc) fraction, and water (H2O) fraction. Among the sub-MICs obtained from resazurin-based fluorimetric microtiter assay, only 50 μg/mL was utilized in evaluating the anti-QS properties of crude extract and fr@*RESULTS@# In the swarming motility assay, hexane fraction (9.39 mm ± 0.67) and DCM fraction (10.82 mm ± 0.95) displayed restriction in the treated P. aeruginosa ATCC 27853 swarms against the control (16.20 mm ± 2.55). In the anti-pyocyanin production assay, hexane fraction exhibited an inhibition of 42.66 % ± 12.94. TLC analysis and phytochemical screening revealed that hexane fraction contains steroids, terpenes, triterpenes, and glycolipids; and DCM fraction contains cardiac glycosides, triterpenoids, terpenes, triterpenes, steroids, alkaloids, and glycolipids.@*CONCLUSION@#Hexane and DCM fractions obtained from A. bilimbi significantly inhibited swarming of P. aeruginosa ATCC 27853 while none of the extracts were able to significantly inhibit pyocyanin formation of P. aeruginosa ATCC 27853.

A case of mass occurrence of Sandalodesmus araujoi () in a municipality of São Paulo, Brazil and description of the heretofore unknown female (Polydesmida, Chelodesmidae) / Um caso de ocorrência em massa de Sandalodesmus araujoi () em um município de São Paulo, Brasil e descrição da fêmea até então desconhecida (Polydesmida, Chelodesmidae)

Abstract Somatic characters are shared by many Chelodesmidae groups, and generic placement and species identifications traditionally have been based on gonopodal morphology. Female genitalic characters have been largely neglected and are rarely photographed or illustrated. This is rather unfortunate as the morphology of female genitalia presents important characters and may be decisive for developing a more robust family classification. We describe the heretofore unknown female of Sandalodesmus araujoi (Schubart, 1946), previously known only from the male holotype collected in São Paulo, Brazil in December 1943; discuss the utility of female genitalic characters for species delineation in Sandalodesmus; and report the first case of a mass occurrence in the Chelodesmidae. While an attempt at a formal diagnosis of Sandalodesmus females based on genitalic characters is premature, the vulvar morphology of the three taxa examined in this study suggests that female genitalia are species-specific. Some characters (i.e., asymmetric valves, presence of digitiform projections and reduction of setae on the internal basal portion of the valves) are constant between the species, suggesting utility for generic-level delineation. Mass occurrences of millipedes are typically unpredictable and likely related to variations in environmental conditions and/or anthropogenic modifications of natural habitats. Although the mass occurrence of S. araujoi reported herein was only observed once, the event coincides with the mating period of millipedes during the rainy season in Brazil. On the other hand, the region where the species was found has been the target of intense urban development, including replacement of natural habitats with residential areas, which may have influenced its population dynamics.

Resumo Os caracteres somáticos são compartilhados por muitos grupos de Chelodesmidae, e o posicionamento genérico e as identificações de espécies tradicionalmente têm sido baseadas na morfologia do gonópodo. Carateres genitais das fêmeas foram amplamente negligenciados e raramente são fotografados ou ilustrados. Isso é lamentável, pois a morfologia da genitália feminina apresenta características importantes e pode ser decisiva para o desenvolvimento de uma classificação mais robusta. Neste trabalho, descrevemos a até então desconhecida fêmea de Sandalodesmus araujoi (Schubart, 1946), anteriormente conhecida apenas pelo holótipo macho coletado em São Paulo, Brasil, em dezembro de 1943; discutimos a utilidade de caracteres genitais femininos para delineamento de espécies em Sandalodesmus; e relatamos o primeiro caso de ocorrência em massa para Chelodesmidae. Embora uma tentativa de diagnose formal para fêmeas de Sandalodesmus com base em caracteres genitais seja prematura, a morfologia vulvar dos três táxons examinados neste estudo, sugere que a genitália feminina é espécie-específica. Alguns caracteres (e.g. válvulas assimétricas, presença de projeções digitiformes e redução de cerdas na margem interna das válvas) são constantes entre as espécies do gênero, sugerindo utilidade para delineamento em nível genérico. Ocorrências em massa de milípedes são tipicamente imprevisíveis e provavelmente relacionadas a variações nas condições ambientais e/ou modificações antropogênicas de habitats naturais. Embora a ocorrência em massa de S. araujoi aqui relatada tenha sido observada apenas uma vez, o evento coincide com o período de reprodução dos milípedes durante a estação chuvosa no Brasil. Por outro lado, a região onde a espécie foi encontrada tem sido alvo de intenso desenvolvimento urbano, incluindo substituição de habitats naturais por áreas residenciais, o que pode ter influenciado sua dinâmica populacional.

In vitro quorum quenching activity of eleusine indica crude ethanolic extract against pseudomonas aeruginosa and serratia marcescens

@#<p style="text-align: justify;"><strong>Introduction:</strong> Nosocomial contaminants such as Pseudomonas aeruginosa and Serratia marcescens are increasingly developing resistance to many antibiotics. One of the promising alternatives that may complement, if not substitute, the use of antibiotics is quorum quenching, the process of interfering with chemical signals that mediate communication between microorganisms. Eleusine indica, a ubiquitous grass used traditionally to treat infections, has been shown to contain metabolites, such as fatty acid derivatives and p-coumaric acid, capable of quorum quenching. To date, there has been no study on the quorum quenching activity of E. indica.</p><p style="text-align: justify;"><strong>Objectives:</strong> This study aimed to determine the in vitro activity of crude ethanolic extract of E. indica leaves against selected quorum-sensing regulated virulence factors of P. aeruginosa and S. marcescens.</p><p style="text-align: justify;"><strong>Methodology:</strong> E. indica leaves were collected, washed, air-dried, and homogenized. Following ethanolic extraction and rotary evaporation, the extract was screened for antimicrobial activity through disk diffusion test and broth microdilution assay. The quorum quenching activity of the extract against P. aeruginosa was measured through swarming motility assay, while the activity against S. marcescens was measured through swarming motility and pigment inhibition assays. The quorum quenching assays were conducted in triplicates, and analysis of variance (ANOVA) was performed to identify differences among the treatment groups.</p><p style="text-align: justify;"><strong>Results:</strong> Disk diffusion test revealed that no zones of inhibition formed against both P. aeruginosa and S. marcescens for varying concentrations of up to 200 mg/mL of the crude extract. Likewise, the MIC of the extract against both P. aeruginosa and S. marcescens was determined to be >200 mg/mL. However, it was shown that the extract, at 50 mg/mL, has statistically significant activity (p<0.05) against the swarming motility of P. aeruginosa, and it is 71.6% as effective in reducing the swarming area of the bacteria compared to cinnamaldehyde. This was not observed when the extract was tested against the swarming motility of and pigment production by S. marcescens.</p><p style="text-align: justify;"><strong>Conclusion:</strong> In this study, the quorum quenching activity of the crude ethanolic extract of E. indica leaves was found to be effective against P. aeruginosa but not against S. marcescens. The compounds that will be identified by further studies may conceivably be used as an adjunct therapy in P. aeruginosa infections and as coating agents in medical devices.</p>

Description of mating behavior, life cycle, and antennal sensilla of Cyclocephala putrida Burmeister, 1847 (Coleoptera, Scarabaeidae, Dynastinae) / Descrição do comportamento de cópula, ciclo de vida e sensilas antenais de Cyclocephala putrida Burmeister, 1847 (Coleoptera, Scarabaeidae, Dynastinae)

Abstract The genus Cyclocephala is common in Brazil (Coleoptera, Scarabaeidae, Dynastinae). The adults of some species are important pollinators, and the larvae develop in the soil, feed on organic matter, and contribute to nutrient cycle, but immatures of some species feed on plant roots, and some were registered causing damage in crops. The mating process of some phytophagous scarab beetles has a chemical recognition step, and the antenna is the main structure involved in the detection of odorants associated with insect communication. In the present study the mating behavior, life cycle, and antennal sensilla of C. putrida are described. The study was conducted at the Universidade Estadual de Mato Grosso do Sul, Cassilândia, Brazil. Adults were collected by a light trap installed from January 2016 to December 2017 and were taken to the laboratory for studies. Adults swarms are brief and were registered from January to February, and specimens were mostly collected at 20:00 to 22:00h. Chemical recognition may occur at least during one of the mating steps, during which the couple kept their antennae moving and the lamellae open, while females select males. In laboratory, the mating process lasted 7.5 minutes on average. The antennae of females have about 3399 sensilla and males about 4229 sensilla. Sensilla placodea types I, II, and III are the most abundant, and sensilla ampullacea, basiconica, and coeloconica are also present. The embryonic period lasted 16.0 days; first, second and third instars lasted 16.0, 48.3, and 165.3 days, respectively. The pupal period lasted 24.0 days. The period between egg deposition and adult emergency is about 271.5 days.

Resumo O gênero Cyclocephala é comum no Brasil (Coleoptera, Scarabaeidae, Dynastinae). Os adultos de algumas espécies são importantes polinizadores, e a larva desenvolve-se no solo, alimenta-se de matéria orgânica e contribui para a ciclagem de nutrientes, mas imaturos de algumas espécies alimentam-se de raízes de plantas, e alguns são registrados causando danos em plantas cultivadas. O processo de cópula de algumas espécies de Scarabaeidae fitófagos, apresentam reconhecimento químico, e nas antenas aparecem várias estruturas responsáveis pela detecção dos odores envolvidos na comunicação. No presente trabalho o comportamento de cópula, ciclo de vida e sensilas antenais de Cyclocephala putrida são descritos. O estudo foi conduzido na Universidade Estadual de Mato Grosso do Sul, Cassilândia, Brasil. Adultos foram coletados com armadilha luminosa instalada de Janeiro de 2016 a Dezembro de 2017, e levados para laboratório para estudos. Os adultos revoam por curto período de janeiro a fevereiro, e os espécimes foram coletados em maior quantidade das 20:00 às 22:00 h. O reconhecimento químico possivelmente ocorre durante as etapas que envolvem o comportamento de cópula, no qual o casal mantém as antenas em movimento e as lamelas abertas e as fêmeas selecionam os machos. Em laboratório a cópula dura 7,5 minutos em média. As antenas das fêmeas possuem 3399 sensilas e os machos 4229 sensilas, e as sensilas placódeas dos tipos I, II e III, foram as mais abundantes e sensilas ampuláceas, basicônicas e coelocônicas também estão presentes. O estágio de ovo durou 16,0 dias; o primeiro, segundo e terceiro instar duraram 16,0, 48,3 e 165,3 dias, respectivamente. O período pupal durou 24,0 dias. O período entre deposição dos ovos e emergência dos adultos é de 271,5 dias em média.

Regulação da expressão de pioverdina dependente de contato em pseudomonas aeruginosa / Regulation of surface-dependent pyoverdine expression in Pseudomonas aeruginosa

A gama-proteobactéria Pseudomonas aeruginosa é um patógeno oportunista humano frequentemente associado a pacientes com queimadura grave e aos portadores de fibrose cística. O estabelecimento de infecção depende de uma série de fatores que contribuem para a virulência deste patógeno, dentre eles a produção de sideróforos e outros sistemas de captação de ferro. Pioverdina é o principal sideróforo sintetizado por bactérias do gênero Pseudomonas e linhagens deficientes na sua produção são incapazes de estabelecer infecção em modelos animais. A regulação da biossíntese deste sideróforo envolve a agregação entre as células, indicando a dependência de contato para completa indução da sua produção. O contato com uma superfície altera o comportamento das células e diversos fenótipos são dependentes deste sinal mecânico. PrlC é uma oligopeptidase A putativamente envolvida na degradação de peptídeo-sinais e PA14_00800, uma pequena proteína com domínio de função desconhecida, codificada por um gene imediatamente à jusante de prlC. Existem poucos trabalhos na literatura sobre PrlC e seus homólogos e nenhuma informação sobre PA14_00800. Este trabalho teve como objetivo elucidar o envolvimento de PrlC e PA14_00800 na regulação da produção de pioverdina por células em contato com uma superfície. Para estabelecer uma correlação na expressão destes genes, um estudo da organização gênica foi realizado por RT-PCR, confirmando que eles fazem parte do mesmo operon e, portanto, que a expressão destes genes é regulada pelos mesmos fatores. Ensaios classicamente modulados pelo segundo mensageiro c-di-GMP, como formação de biofilme e motilidade, não apresentaram variações nas linhagens mutantes ΔprlC, ΔPA14_00800 ou Δoperon, indicando que a deleção destes genes não altera significativamente os níveis de c-di-GMP nas células. A motilidade do tipo swarming é, no entanto, severamente afetada na linhagem ΔPA14_00800 quando o meio de cultura não contém cloreto de cálcio e glicose, indicando um defeito na sinalização celular ou requerimento energértico desta linhagem nestas condições. PA14_00800 regula a fluorescência de P. aeruginosa em meio sólido e semissólido, mas não em meio líquido. Esta fluorescência depende tanto de pioverdina quanto de PQS, umamolécula de comunicação celular fluorescente, e a possibilidade de outros fatores estarem envolvidos neste fenótipo ainda está sob investigação. Análise do transcritoma por RNASeq com a linhagem ΔPA14_00800 comparada à linhagem parental foi realizada a partir de colônias destas linhagens crescidas em M9 modificado. Genes envolvidos no sistema de secreção do tipo III e do tipo VI e na biossíntese de PQS apareceram dentre os genes diferencialmente expressos, bem como genes para o catabolismo de glicose. Este trabalho foi o primeiro a investigar o papel de PA14_00800 na fisiologia de P. aeruginosa, e os conhecimentos adquiridos aqui podem ser transpostos, com cautela, para compreensão da função dos homólogos de PA14_00800 em outras bactérias

The gamma-proteobacterium Pseudomonas aeruginosa is a human opportunistic pathogen frequently associated with patients with severe burns and those with cystic fibrosis. The establishment of infection depends on several factors that contribute to the virulence of this pathogen, among them siderophore production and other iron uptake systems. Pyoverdine is the main siderophore synthesized by the bacteria of the genus Pseudômonas and pyoverdinedeficient strains are unable to establish infection in animal models. The regulation of biosynthesis of this siderophore involves cell aggregation, indicating contact dependency for complete induction of pyoverdine production. Surface contact alters cell behavior and several phenotypes are dependent on this mechanical cue. PrlC is an oligopeptidase A putatively involved in peptide-signals degradation and PA14_00800, a small protein with a domain of unknown function, encoded by a gene immediately downstream of prlC. There are few papers in the literature on PrlC and its homologues and no information on PA14_00800. This work aimed to elucidate the role of PrlC and PA14_00800 in surface-dependent regulation of pyoverdine production. To establish a correlation in the expression of these genes, a study of the gene organization was performed by RT-PCR, confirming that they are part of an operon and therefore the expression of these genes is regulated by the same factors. Traits classically modulated by the second messenger c-di-GMP, such as biofilm formation and motility, did not show variations in the ΔprlC, ΔPA14_00800 or Δoperon, indicating that the deletion of these genes does not significantly alter the levels of c-di-GMP within the cells. Swarming motility is, however, severely affected in the strain ΔPA14_00800 when the culture medium does not contain calcium chloride and glucose, indicating a cell signaling defect or energetic requirement under these conditions. PA14_00800 regulates surface-dependent fluorescence of P. aeruginosa, in solid and semi-solid medium. This fluorescence depends on both pyoverdine and PQS, a fluorescent cell-to-cell communication molecule, and the investigation of other putative factors involved in this phenotype is still under study. Transcriptomic analysis by RNASeq with the strain ΔPA14_00800 compared to PA14 was performed from colonies ofthese strains grown in modified M9 1% agar. Genes involved in the type III and type VI secretion systems, in PQS biosynthesis and glucose catabolism were differentially expressed. This work was the first to investigate the role of PA14_00800 in the physiology of P. aeruginosa, and the knowledge obtained here can be cautiously transposed to understanding the role of PA14_00800 homologues in other bactéria

Mechanisms and molecular typing of carbapenem-resistant Proteus mirabilis strains deficient in swarming motility / 中华微生物学和免疫学杂志

Objective To investigate the mechanism and epidemiological characteristics of carbap-enem-resistant Proteus mirabilis ( PM) strains deficient in swarming motility. Methods PM strains were isolated from Hangzhou General Hospital of CAPF ( Chinese People′s Armed Police Forces) during January 2013 to December 2014. Bacterial motility and flagella of the PM strains were observed through semi-solid agar culture and flagella staining. Pulsed-field gel electrophoresis ( PFGE) was performed for homology anal-ysis. Antimicrobial susceptibility test and phenotypic confirmatory test were also carried out. PCR analysis and DNA sequencing were performed to confirm the genotype of resistant genes. Plasmid electroporation and S1-PFGE in combination with Southern blot hybridization were used to determine the location of the carbap-enem-resistant genes. Genetic structure of the blaKPC-2 gene was obtained by PCR mapping. Results A total of 42 PM isolates deficient in swarming motility were screened out and the resistance rates to imipenem and meropenem were 57. 1% and 52. 4%, respectively. PCR analysis and DNA sequencing confirmed that 24 carbapenem-resistant PM isolates deficient in swarming motility carried blaKPC-2 gene and belonged to three clones as indicated by the results of PFGE. Southern blot hybridization indicated that the blaKPC-2 gene was located on plasmids varying in size (26 kb, 55 kb and 139 kb). In addition, some of the strains harbored several resistant genes, such as blaTEM-1 , blaCTX-M-65 and rmtB. The genetic structures of strains carrying blaKPC-2 gene were ISKpn8, blaKPC-2 and ISKpn6-like from upstream to downstream. Conclusion Compared with the PM strains with swarming motility, the carbapenem-resistance rate was significantly higher in these PM strains deficient in swarming motility. Carbapenemases KPC-2 played an important role in the carbapen-em-resistant PM strains deficient in swarming motility. There was a cloning spread trend for carbapenem-re-sistant PM strains in our hospital. Clinicians should pay more attention to the risk of spreading.

Phenolic compounds affect production of pyocyanin, swarming motility and biofilm formation of Pseudomonas aeruginosa

Objective: To investigate the effects of plant-derived phenolic compounds (i.e. caffeic acid, cinnamic acid, ferulic acid and vanillic acid) on the production of quorum sensing regulated virulence factors such as pyocyanin, biofilm formation and swarming motility of Pseudomonas aeruginosa (P. aeruginosa) isolates. Methods: Fourteen clinical P. aeruginosa isolates obtained from urine samples and P. aeruginosa PA01 strain were included in the study. The antibacterial effects of phenolic compounds were screened by well diffusion assay. Pyocyanin and biofilm ac-tivity were measured from culture supernatants and the absorbance values were measured using a spectrophotometer. Swarming plates supplemented with phenolic acids were point inoculated with P. aeruginosa strains and the ability to swarm was determined by measuring the distance of swarming from the central inoculation site. Results: Tested phenolic compounds reduced the production of pyocyanin and biofilm formation without affecting growth compared to untreated cultures. Moreover, these compounds blocked about 50% of biofilm production and swarming motility in P. aeruginosa isolates. Conclusions: We may suggest that if swarming and consecutive biofilm formation could be inhibited by the natural products as shown in our study, the bacteria could not attach to the surfaces and produce chronic infections. Antimicrobials and natural products could be combined and the dosage of antimicrobials could be reduced to overcome antimicrobial resistance and drug side effects.

Phenolic compounds affect production of pyocyanin, swarming motility and biofilm formation of Pseudomonas aeruginosa

Objective To investigate the effects of plant-derived phenolic compounds (i.e. caffeic acid, cinnamic acid, ferulic acid and vanillic acid) on the production of quorum sensing regulated virulence factors such as pyocyanin, biofilm formation and swarming motility of Pseudomonas aeruginosa (P. aeruginosa) isolates. Methods Fourteen clinical P. aeruginosa isolates obtained from urine samples and P. aeruginosa PA01 strain were included in the study. The antibacterial effects of phenolic compounds were screened by well diffusion assay. Pyocyanin and biofilm activity were measured from culture supernatants and the absorbance values were measured using a spectrophotometer. Swarming plates supplemented with phenolic acids were point inoculated with P. aeruginosa strains and the ability to swarm was determined by measuring the distance of swarming from the central inoculation site. Results Tested phenolic compounds reduced the production of pyocyanin and biofilm formation without affecting growth compared to untreated cultures. Moreover, these compounds blocked about 50% of biofilm production and swarming motility in P. aeruginosa isolates. Conclusions We may suggest that if swarming and consecutive biofilm formation could be inhibited by the natural products as shown in our study, the bacteria could not attach to the surfaces and produce chronic infections. Antimicrobials and natural products could be combined and the dosage of antimicrobials could be reduced to overcome antimicrobial resistance and drug side effects.

Motility of Bacterial Pathogens in Minimal Media Containing Human Serum.

Aims: To investigate the effect of human serum on growth pattern, cellular morphology, and motility of potentially pathogenic bacteria. Study Design: This was an analytic experimental study. Place and Duration of Study: Institute of Exact Sciences and Technology (ICET), Federal University of Amazonas (UFAM), Mycology Laboratory, between August 2012 and July 2013. Methodology: Growth of Escherichia coli, Salmonella typhi, Bacillus subtilis and Bacillus cereus was examined on hard agar (1.5%) solid medium containing 0, 20, or 40 percent pooled human serum. Dilutions of each species were point-inoculated at the center of the plate. Cultivation was carried out aerobically for up to 20 days at 37°C in sealed humidified boxes. Spreading growth was examined by measuring colony diameters, analyzing macroand micromorphology, and measuring the fractal dimension of colonies. Results: E. coli and S. typhi strains grew better in Davis and Mingioli agar, whereas B. subtilis and B. cereus grew better in Fujikawa agar. B. cereus and S. typhi developed a white halo of proteolysis around the colony in the medium supplemented with serum. The addition of human serum to minimal hard-agar medium induced a cellular phenotypic change and a colony morphological change, especially in B. cereus and S. typhi. B. cereus and S. typhi developed elongated cells on the colony edge in the presence of human serum, showing cells in raft-like association. Generally, colonies of bacteria grown in the absence of human serum presented smaller fractal and growth dimensions and morebranched spreading, presumably in a sliding translocation. Conclusion: Cells of “temperate swarmer” species translocated more efficiently on hard agar supplemented with human serum, by sliding and possibly by swarming. The presence of human blood or serum, despite the inhibitory activity of antibodies, may allow pathogenic bacterial cells to overcome the difficulties of low levels of nutrients and hard surfaces with little available water, and may facilitate translocation to other sites. Further investigations of the influence of human serum on swarming and sliding are warranted.

Isolation, identification & characterization of Proteus penneri - a missed rare pathogen.

Background & objectives: Indole negative Proteus species are invariably incorrectly identified as P. mirabilis, missing isolates of Proteus penneri. P. penneri is an invasive pathogen capable of causing major infectious diseases still seldom reported in individual cases. We report here the isolation, differentiation, characterization and typing of P. penneri from patients with different clinical infections. Methods: Urine, pus and body fluids collected from patients in intensive care units, wards and out patients departments of a tertiary health care institute from north India were cultured. A total of 61 indole negative Proteus isolates were subjected to extended biochemical tests to differentiate and identify P. penneri from P. mirabilis including failure to produce ornithine decarboxylase (by 0% strains of P. penneri and 100% strains of P. mirabilis) besides P. penneri being uniformly salicin negative, non-utilizer of citrate but ferments sucrose and maltose. Antibiograms and Dienes phenomenon were performed to characterize and type P. penneri isolates besides screening for β-lactamase production. Results: Eight isolates of P. penneri were identified; four from urine, three from abdominal drain-fluid and one from diabetic foot ulcer. P. penneri was isolated as the sole pathogen in all patients having underlying disease; post-operatively. Swarming was not seen in the first strain on primary isolation and was poor in strain-4. All eight isolates were biochemically homologous but multi-drug resistant (MDR) with resistance to 6-8 drugs (up to 12). β-lactamase production was seen in three of five isolates while Dienes phenomenon found four distinct types and discriminated strains differing in resistance even with a single drug. Interpretation & Conclusions: A few additional biochemical tests identified P. penneri isolates; it infected patients with underlying disease and strains were MDR and heterogenous.

Effects of Pseudomonas aeruginosa pvdQ gene on altering antibiotic susceptibility under swarming conditions / 中华微生物学和免疫学杂志

Objective To investigate whether Pseudomonas aeruginosa pvdQ( PA2385 ) gene reveals altered antibiotic susceptibility under swarming conditions. Methods The plasmid pME6032 with pvdQ gene was constructed and identified, then transformed into Pseudomonas aeruginosa PAO1 by the electroporation, building pvdQ overexpression strain. Using the same method building pME6032-PAO1 strain.Bacteria were inoculated in LB overnight , measuring the colony diameter of the swarming zone . Results Strains of pvdQ overexpression was successfully constructed by real-time PCR. Comparison of two strains of the swarming motility of change in diameter: The result showed that PAO1 and pvdQ overexpression strains can both improve the antibiotics resistance. Swarmer cells of pvdQ overexpression strain exhibited a 2- to 4-fold increase in antibiotic resistance toward ceftazidime,ciprofloxacin, meropenem and polymyxin B compared to PAO1 on BM2-swarming agar plates. Conclusion pvdQ gene played an important role in elevating the antibiotics resistance, which through prarticipated in the swarmer cell differentiation.

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

Periodic variety in swarming motility of Proteus mirabilis / 第三军医大学学报

Objective To investigate the periodic variety at the germ form, quantity, flagellar expression and activity of breath enzyme etc during the swarming motility of Proteus mirabilis (PM). Methods The 2, 3, 5-tetraphenyltetrazolium chloride (TTC), as indictor of breath enzyme, was added to the culture medium. Electronic microscope was adopted to observe the germ form and flagellar. Quantity of PM was measured by turbidimetry. Results During the PM growth and migration at LB medium, the germ quantity reduced and form changed from short and small to long and thin, flagellar got more and breath enzyme turned into inactive. Conclusion At the LB medium, PM in growth process appeared varieties in length of germ body, flagellar quantity and germs density, etc. PM in solid medium appeared wave motility, resulting from many factors including the environment, the signal transduction in cells and the density of germ.

Abnormal flagellar production in O-antigen mutants of Citrobacter freundii / 第三军医大学学报

Objective To study the molecular mechanism of swarming motility in Citrobacter freundii.Methods Mini-Tn5 transposon was used to induce swarming mutants in Citrobacter freundii.The mutant genes were amplified by inverse-PCR and were then sequenced.Mutants were observed under light microscope and electron microscope to determine the characters of motility and bacterial shape.Results Seven swarming mutants were found to be three gene mutants(wzxB,waaW,waaL) involving LPS synthesis.Moreover,flagellar production of these mutants was abnormal,that is,only small portion of bacterial cells produced flagella normally.The majority of cells tended to aggregate and produced little flagella observed under electron microscope.Conclusion Absence of O-antigen affects the production of flagella and swarming motility of Citrobacter(freundii).