Proteus vulgaris and Proteus mirabilis Decrease Candida albicans Biofilm Formation by Suppressing Morphological Transition to Its Hyphal Form.

PURPOSE: Candida albicans (C. albicans) and Proteus species are causative agents in a variety of opportunistic nosocomial infections, and their ability to form biofilms is known to be a virulence factor. In this study, the influence of co-cultivation with Proteus vulgaris (P. vulgaris) and Proteus mirabilis (P. mirabilis) on C. albicans biofilm formation and its underlying mechanisms were examined. MATERIALS AND METHODS: XTT reduction assays were adopted to measure biofilm formation, and viable colony counts were performed to quantify yeast growth. Real-time reverse transcriptase polymerase chain reaction was used to evaluate the expression of yeast-specific genes (rhd1 and rbe1), filament formation inhibiting genes (tup1 and nrg1), and hyphae-related genes (als3, ece1, hwp1, and sap5). RESULTS: Candida biofilm formation was markedly inhibited by treatment with either living or heat-killed P. vulgaris and P. mirabilis. Proteus-cultured supernatant also inhibited Candida biofilm formation. Likewise, treatment with live P. vulgaris or P. mirabilis or with Proteus-cultured supernatant decreased expression of hyphae-related C. albicans genes, while the expression of yeast-specific genes and the filament formation inhibiting genes of C. albicans were increased. Heat-killed P. vulgaris and P. mirabilis treatment, however, did not affect the expression of C. albicans morphology-related genes. CONCLUSION: These results suggest that secretory products from P. vulgaris and P. mirabilis regulate the expression of genes related to morphologic changes in C. albicans such that transition from the yeast form to the hyphal form can be inhibited.

Production of acylated homoserine lactone by a novel marine strain of Proteus vulgaris and inhibition of its swarming by phytochemicals.

A marine strain of Proteus vulgaris capable of activating multiple acylated homoserine lactone (AHL)-based reporter cultures was isolated. The cognate signal molecule was characterized as octanoyl homoserine lactone (OHL) and its production was observed to be growth dependent, with maximum production (5.675 µg l(-1)) at 24 h growth. The strain exhibited swarming, but its motility was not affected upon addition of pure OHL or culture supernatant. Phytochemicals such as quercitin and berberine chloride inhibited OHL production and reduced swarming. FliA, the predominantly upregulated protein during swarming, was considered as a possible target for these inhibitors, and docking of the two most active and two least active inhibitors to this protein suggested preferential binding of the former set of compounds. Apart from adding new evidence to AHL production in Proteus vulgaris, active inhibitors shortlisted from this study could help in identifying lead compounds to act against this opportunistic pathogen of the respiratory and gastrointestinal tract.

Influence of bacterial presence on biofilm formation of Candida albicans.

PURPOSE: Candida albicans is an opportunistic pathogen that is commonly found in human microflora. Biofilm formation (BF) is known as a major virulence factor of C. albicans. The aim of this study was to examine the influence of bacterial presence on biofilm formation of C. albicans. MATERIALS AND METHODS: The BF of Candida was investigated when it was co-cultured with C. albicans (C. albicans 53, a yeast with a low BF ability, and C. albicans 163, a yeast with high BF ability) and bacteria. BF was assessed with XTT reduction assay. A scanning electron microscope was used to determine the structure of the biofilm, and real-time reverse transcriptase polymerase chain reaction was used to amplify and quantify hyphae-associated genes. RESULTS: Co-culturing with two different types of bacteria increased the BF value. Co-culturing with C. albicans 53 and 163 also increased the BF value compared to the value that was obtained when the C. albicans was cultured individually. However, co-culturing with bacteria decreased the BF value of C. albicans, and the BF of C. albicans 163 was markedly inhibited. The expression of adherence and morphology transition related genes were significantly inhibited by co-culturing with live bacteria. CONCLUSION: Bacteria have a negative effect on the formation of biofilm by C. albicans. This mechanism is the result of the suppression of genes associated with the hyphae transition of C. albicans, and bacteria particles physically affected the biofilm architecture and biofilm formation.

[Antiadhesive potencial of Rhodococcus erythropolis IMB Ac-5017 biosurfactants].

The effect of Rhodococcus erythropolis IMB Ac-5017 biosurfactants (surface-active substances, SAS) with different degree of purification on attachment of bacteria (Escherichia coli IEM-1, Bacillus subtilis BT-2, Proteus vulgaris BT-1, Staphylococcus aureus BMC-1, Pseudomonas aeruginosa P-55, Enterobacter cloacae AC-22, Erwinia aroidaeae B-433), yeasts (Candida albicans D-6) and fungi (Aspergillus niger P-3, Fusarium culmorum T-7) to the abiotic surfaces (glass, plastic, ceramics, steel, linoleum) was studied. The dependence of microorganisms adhesion on degree of SAS purification (supernatant, purified SAS solution), SAS concentration (0,04-1,25 mg/ml), type of surface and test-cultures was established. The adhesion of majority investigated bacterial cells after treatment of abiotic surfaces with supernatant of cultural liquid with SAS concentration 0,06-0,25 mg/ml was on the average 20-45, yeasts C. albicans D-6--30-75% and was less than that purified SAS solution with the same concentration. Higher antiadhesive activity of supernatant as compared to purified SAS solution testifies to possibility of exception of the expensive stage of isolation and purification at obtaining of preparations with antiadhesive properties.

[Role of superoxide anion radicals in the bacterial corrosion of metals].

It was found that seven strains of bacteria can cause corrosion damage to aluminum, its alloys, and zinc. With respect to the studied metals, the most active bacteria were Proteus vulgaris 1212 and Pseudomonas aeruginosa 969. Superoxide anion radicals were demonstrated to play a role in the initiation of corrosive damage to aluminum and zinc, while bacterial exometabolites participate in the later stages of this process.

Electron transfer from Proteus vulgaris to a covalently assembled, single walled carbon nanotube electrode functionalised with osmium bipyridine complex: application to a whole cell biosensor.

We report the fabrication and use of electrodes constructed from single walled carbon nanotubes (SWCNTs) chemically assembled on a carbon surface and functionalised with an osmium(II) bipyridine complex (Osbpy). The ability of the electrodes to transduce biologically generated currents from Proteus vulgaris has been established. Our investigations show that there are two contributions to the current: one from electroactive species secreted into solution and another from cell redox sites. The modified electrode can be used to monitor cell metabolism, thereby acting as a whole cell biosensor. The biosensor was used in a 1-h assay to investigate the toxicity of ethanol, sodium azide and the antibiotic ampicillin and gave quantitative data that were closely correlated with standard cell plate viability assays. The results provide proof of principle that the whole cell biosensor could be used for high throughput screening of antimicrobial activity. One of the modified electrodes was used for approximately 1000 measurements over four months demonstrating the robustness of the system.

Crystalline bacterial biofilm formation on urinary catheters by urease-producing urinary tract pathogens: a simple method of control.

The problem of catheter encrustation stems from infection by urease-producing bacteria. These organisms generate ammonia from urea, elevate the pH of urine and cause crystals of calcium and magnesium phosphates to form in the urine and the biofilm that develops on the catheter. In this study, a laboratory model was used to compare the ability of 12 urease-positive species of urinary tract pathogens to encrust and block catheters. Proteus mirabilis, Proteus vulgaris and Providencia rettgeri were able to raise the urinary pH above 8.3 and produce catheter-blocking crystalline biofilms within 40 h. Morganella morganii and Staphylococcus aureus elevated the pH of urine to 7.4 and 6.9, respectively, and caused some crystal deposition in the biofilms but did not block catheters in the 96 h experimental period. Isolates of Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Serratia marcescens, Pseudomonas aeruginosa and Providencia stuartii were only capable of raising the pH of urine to a maximum of 6.4 and failed to cause crystal deposition in the biofilm. The most effective way to prevent catheter encrustation was shown to be diluting urine and increasing its citrate concentration. This strategy raises the nucleation pH (pH(n)) at which calcium and magnesium phosphates crystallize from urine. Increasing the fluid intake of a healthy volunteer with citrated drinks resulted in urine with a pH(n) of >8.0 in which catheter encrustation was inhibited. It is suggested that this dietary strategy will be an effective means of controlling catheter encrustation, whichever bacterial species is causing the problem.

[Effect of the proteolytic enzymes of Bacillus licheniformis and the lysoamidase of Lysobacter sp. XL1 on Proteus vulgaris and Proteus mirabilis].

Preparations of culture liquid of three Bacullus licheniformis strains (S, 103, and 60.4) and the enzymatic preparation lysoamidase from culture liquid of Lysobacter sp. strain XL1 actively lysed preliminarily autoclaved cells of gram-negative bacteria Proteus vulgaris and P. mirabilis. Living Proteus cells treated with these enzymatic preparations were lysed during their subsequent autoclaving. Inoculation of enzyme-treated Proteus cells, taken either separately or in combination with one another and polymyxin B, into a rich medium led to cell repair and restoration of viability of culture.

Membrane fluidity sensoring microbial fuel cell.

A study has been performed to examine the effect of temperature and ethanolic stresses on the coulombic efficiency of a microbial fuel cell. The conventional-type fuel cell containing Gram-negative bacteria, Proteus vulgaris, was investigated as a model system. From current output measurements, it was found that the coulombic yields were altered by environmental stresses such as temperature shock or ethanol treatment to the bacteria. While high-temperature or ethanolic shock led to a remarkable decrement in coulombic output, the low-temperature shock induced a slight increase in microbial fuel cell efficiency. These results indicate that the membrane fluidity is affected considerably by environmental stress, which in turn affects the electron transfer process through the bacterial cell membrane to and from the electrode. This interpretation was confirmed by the cyclic voltammetric study of a mediator on an electrode surface modified with the lipids extracted from the membrane of P. vulgaris under the given stress. Markedly different electrochemical behaviors were observed depending on the environmental stress. A reciprocal relationship between coulomb output and the ratio of saturation/unsaturation of fatty acids has been observed. This is the first report, to our knowledge, that the structural adaptation of membrane fatty acids in response to the environmental shock can regulate the coulombic efficiency of a microbial fuel cell.

Influence of different natural physical fields on biological processes.

In space flight conditions gravity, magnetic, and electrical fields as well as ionizing radiation change both in size, and in direction. This causes disruptions in the conduct of some physical processes, chemical reactions, and metabolism in living organisms. In these conditions organisms of different phylogenetic level change their metabolic reactions undergo changes such as disturbances in ionic exchange both in lower and in higher plants, changes in cell morphology for example, gyrosity in Proteus (Proteus vulgaris), spatial disorientation in coleoptiles of Wheat (Triticum aestivum) and Pea (Pisum sativum) seedlings, mutational changes in Crepis (Crepis capillaris) and Arabidopsis (Arabidopsis thaliana) seedling. It has been found that even in the absence of gravity, gravireceptors determining spatial orientation in higher plants under terrestrial conditions are formed in the course of ontogenesis. Under weightlessness this system does not function and spatial orientation is determined by the light flux gradient or by the action of some other factors. Peculiarities of the formation of the gravireceptor apparatus in higher plants, amphibians, fish, and birds under space flight conditions have been observed. It has been found that the system in which responses were accompanied by phase transition have proven to be gravity-sensitive under microgravity conditions. Such reactions include also the process of photosynthesis which is the main energy production process in plants. In view of the established effects of microgravity and different natural physical fields on biological processes, it has been shown that these processes change due to the absence of initially rigid determination. The established biological effect of physical fields influence on biological processes in organisms is the starting point for elucidating the role of gravity and evolutionary development of various organisms on Earth.

Classification of Proteus vulgaris biogroup 3 with recognition of Proteus hauseri sp. nov., nom. rev. and unnamed Proteus genomospecies 4, 5 and 6.

Strains traditionally identified as Proteus vulgaris formed three biogroups. Biogroup 1, characterized by negative reactions for indole production, salicin fermentation and aesculin hydrolysis, is now known as Proteus penneri. Biogroup 2, characterized by positive reactions for indole, salicin and aesculin, was shown by DNA hybridization (hydroxyapatite method) to be a genetic species separate from biogroup 1 and from biogroup 3 which is positive for indole production and negative for salicin and aesculin. In this study, 52 strains were examined, of which 36 strains were Proteus vulgaris biogroup 3, which included the current type strain of the species P. vulgaris (ATCC 29905T), and compared to seven strains of Proteus vulgaris biogroup 2 and nine type strains of other species in the genera Proteus, Providencia and Morganella. By DNA hybridization, these 36 strains were separated into four distinct groups, designated as Proteus genomospecies 3, 4, 5 and 6. DNAs within each separate Proteus genomospecies were 74-99% related to each other in 60 degrees C hybridization reactions with < or = 4.5% divergence between related sequences. Proteus genomospecies 3 contained the former P. vulgaris type strain and one other strain and was negative in reactions for salicin fermentation, aesculin hydrolysis and deoxyribonuclease, unlike the reactions associated with strains considered as typical P. vulgaris which are positive in reactions for salicin, aesculin and DNase. Genomospecies 3 can be distinguished from Proteus genomospecies 4, 5 and 6 because it is negative for Jordan's tartrate. Proteus genomospecies 4, containing five strains, was differentiated from Proteus penneri, genomospecies 3 and 6 and most, but not all, strains of genomospecies 5, by its ability to ferment L-rhamnose. Proteus genomospecies 5 and 6, containing 18 and 11 strains, respectively, could not be separated from each other by traditional biochemical tests, by carbon source utilization tests or SDS-PAGE of whole-cell proteins. In an earlier publication, a request was made to the Judicial Commission that the former type strain of P. vulgaris (ATCC 13315) be replaced by P. vulgaris biogroup 2 strain ATCC 29905T, a strain considered more biochemically typical of P. vulgaris strains. This would have the effect of assigning the name P. vulgaris to P. vulgaris biogroup 2. Since this request has been acceded to, the name Proteus hauseri is herein proposed for Proteus vulgaris genomospecies 3. Its type strain is ATCC 700826T. Proteus genomospecies 4, 5 and 6 will remain unnamed until better phenotypic differentiation can be accomplished. All Proteus genomospecies were similar in their antimicrobial susceptibility patterns. Nineteen strains were isolated from urine, four from faeces, two from wounds, nine from other human sources and two from animals.

Plant beneficial effect of two strains of Proteus vulgaris isolated from tea plantations.

Two strains of Proteus isolated from tea plantation soil were tested for their ability to colonise the roots of gram (Cicer arietinum), bean (Phaseolus radiatus) and mung (Phaseolus mungo) using a gnotobiotic system. Seeds bacterized with the two strains grew faster and showed significant increase in root and shoot enlargement of the plants tested. The bioactive fractions obtained from the culture filtrates and separated through HPLC showed that the plant growth promoting fractions were not always fungicidal and that the insecticidal fraction which was found only in RRLJ 16 was not plant growth promoting. These results suggest that the plant growth promotion effect of the plant beneficial bacteria may not always be due to disease suppression.

[A probiotic as an antagonist of bacterial translocation in experimental pancreatitis]. / Un probiotico come antagonista della traslocazione batterica nella pancreatite sperimentale.

Infection is the most common cause of death in acute pancreatitis. Earlier studies have demonstrated that early enteral nutrition decreases microbial translocation, upregulates the immune function and reduces septic complications and mortality. Lactobacillus plantarum (Lp) has been shown to be effective in reducing egress of endotoxin and microbial strain that showed very high adherence power to gut mucosa. We adopted a model of acute pancreatitis induced by isolation and ligation of biliopancreatic duct in adult Lewis rats. Three groups were studied: A. control group (sham operation); B. induced pancreatitis, no further treatment; C. Induced pancreatitis + gavage with 5 ml/day of a suspension of Lp 299 v in a dose of 0.5-1.0 x 10(9)/ml during 4 days before and 4 days after induction of pancreatitis. All animals were sacrificed after 96 hours. Histological studies and microbiological analyses were performed. Forty out of 55 animals showed signs of severe pancreatitis on sacrifice after 96 hours. Only these animals were further studied. In group A, we found only 1/20 bacteria in mesenteric nodes (MN). Pathogenic microrganisms were found in the non-treated group in MN in 14/20 and in the pancreatic tissue in 10/20. In contrast, when kept on an umbrella of Lp 299 v, only 4/20 animals demonstrated growth of enteric bacteria in MN and 3/20 in pancreatic tissue. All of these results showed a significant reduction of infection in the treated groups. In our model, Lp 299 v is effective in preventing microbial translocation in experimental pancreatitis. Treatment with probiotic bacteria, such as Lactobacillus spp, seems to be a promising alternative as problems with antibiotic-resistant bacteria seem to accumulate.

Potential virulence factors of Proteus bacilli.

The object of this review is the genus Proteus, which contains bacteria considered now to belong to the opportunistic pathogens. Widely distributed in nature (in soil, water, and sewage), Proteus species play a significant ecological role. When present in the niches of higher macroorganisms, these species are able to evoke pathological events in different regions of the human body. The invaders (Proteus mirabilis, P. vulgaris, and P. penneri) have numerous factors including fimbriae, flagella, outer membrane proteins, lipopolysaccharide, capsule antigen, urease, immunoglobulin A proteases, hemolysins, amino acid deaminases, and, finally, the most characteristic attribute of Proteus, swarming growth, enabling them to colonize and survive in higher organisms. All these features and factors are described and commented on in detail. The questions important for future investigation of these facultatively pathogenic microorganisms are also discussed.

The effect of granulocyte colony-stimulating factor (G-CSF) on bacterial translocation in the splenectomized rat.

BACKGROUND/AIMS: We created a study group in order to investigate the effects of splenectomy and filgrastim. Filgrastim is an immunomodulator granulocyte colony-stimulating factor (G-CSF), that affects bacterial translocation. MATERIALS AND METHODS: We created 3 study groups with 30 male Sprague-Dawley rats; the first group included sham splenectomy, the second group was splenectomy, and the third group was splenectomy+ filgrastim group. RESULTS: The mean bacterial colony count of the cecum were 2.5 x 10(9) in group 1, 1.2 x 10(10) in group 2 and 3.5 x 10(9) in group 3. The differences between these groups were accepted as statistically significant. The mean counts of the terminal ileum were 1.1 x 10(9) in group 1, 5.5 x 10(10) in group 2 and 2.5 x 10(10) in group 3. The p values of group 1-2 were 0.036 (statistically significant) and 0.123 in groups 2-3) were not statistically significant. The mean counts of the liver were 0.2 x 10(4), 1 x 10(10) and 3.4 x 10(5), respectively. In comparison of the groups the p values of the first and last 2 groups were found to be 0.047 (statistically significant). The mean counts of the mesenteric lymph node were 0.7 x 10(3), 1 x 10(10) and 0.9 x 10(6) respectively. The p values were 0.343 for the first and the last 2 groups both. As they were above 0.05, they were not statistically significant. The degrees of liver Kupffer cell hyperplasia were (+) 40%, (+2) 50% and (+3) 10% (group 1), (+) 10%, (+2) 40% and (+3) 50% (group 2), (+2) 60% and (+3) 40% (group 3) (p = 0.0039). The rates of the absence of pathology in mesenteric lymph nodes were 70% (group 1), 90% (group 2) and 100% (group 3) (p = 0.049). These findings were statistically significant. CONCLUSIONS: We found that splenectomy has activated the whole predisposing factors of bacterial translocation and created the latter itself. In addition, we showed that filgrastim, a recently widespread used G-CSF, decreases bacterial translocation significantly.

Examination of biofilm formation and risk of infection associated with the use of urinary catheters with leg bags.

Urinary catheters and legs bags were simultaneously colonized by Escherichia coli and Proteus vulgaris using a model urinary drainage system. the system was continuously supplied with filter-sterilized artificial urine using a diurnal flow pattern. The extent of colonization was determined by assessment of both planktonic and biofilm formation over time. Contamination of the catheters resulted in rapid colonization of the whole system within a 24 h period. Contamination of the leg bags resulted in an ascending biofilm formation over a four-day period. Results indicated that infection risk could be minimized by changing the catheter and leg bags at least once a week. The design of the leg bags was not found to influence the rate or extent of biofilm formation.

Mechanisms of antimotility action of tricyclic compounds in Proteus vulgaris.

Tricyclic compounds were able to inhibit the motility of Proteus vulgaris. The effectiveness of antimotility action was related to the physicochemical properties of the molecules, i.e. energy of HOMO, Log P, total surface. The antimotility action of the compounds was due to their reversible inhibition on the proton pump of the bacterium. Phosphate anion antagonized the antimotility, and potassium cation enhanced the action of phosphate anion on the antimotility effect induced by the agents. Glucose reversed the antimotility action of the compounds. Factors directly increasing the bacterial proton-motive force (PMF) could change bacterial motility and the antimotility action of the tricyclic compounds.

Effects of some tricyclic psychopharmacons and structurally related compounds on motility of Proteus vulgaris.

A simple test for the evaluation of drugs interfering with bacterial motility was established with Proteus vulgaris. With this model, promethazine, 7-hydroxy-chlorpromazine, imipramine, 7,8-dioxochlorpromazine and acridine orange were shown to exert significant motility and swarming inhibitory action on Proteus vulgaris strains at subinhibitory concentrations. Quinidine enhanced the antimotility effect of promethazine. The antimotility effect of promethazine was synergized by proton pump inhibitors omeprazole and abscissic acid, but antagonized by extracellular potassium and sodium ions.

The Dienes effect as an epidemiological tool in a paraplegic unit.

The authors study the Dienes effect in 136 P.mirabilis and 2 P.vulgaris strains, isolated from different infection or colonization sites, in 27 spinal cord injured patients, admitted to a Paraplegic Unit, during a 9 month period. It is shown that 7 cross-infection and 18 cross-contamination episodes occurred, affecting 13 of 27 patients, which indicates the great spreading capacity of these microorganisms; 97% of the results obtained were observed once again after 6 months' storage of strains. In addition, in 7 of these strains their resistance to several antibiotics was cured by treating them with acriflavine salts, demonstrating that the Dienes effect persists even when resistance to antibiotics is modified; the authors conclude that the Dienes effect is an accurate stable epidemiological tool to identify cross-infections and its origins, and to facilitate the interruption of the chain of infection.