Antimicrobial activity of berberine--a constituent of Mahonia aquifolium

The antimicrobial activity of the protoberberine, alkaloid, berberine, isolated fromMahonia aquifolium, was evaluated against 17 microorganisms including two Gram-negative bacteria—Pseudomonas aeruginosa andEscherichia coli (both resistant and sensitive), two Gram-positive bacteria—Bacillus subtilis andStaphylococcus aureus, Zoogloea ramigera, six filamentous fungi—Penicilium chrysogenum, Aspergillus niger, Aureobasidium pullulans (black and white strain),Trichoderma viride (original green strain and brown mutant),Fusarium nivale, Mycrosporum gypseum, and two yeasts—Candida, albicans andSaccharomyces cerevisiae. The IC₅₀, minimum inhibitory concentration (MIC), minimum microbicidal concentration (MMC) and minimum microbistatic concentration (MMS) varied considerably depending on the microorganism tested, the sensitivity decreasing as follows:S. aureus >P. aeruginosa S (sensitive) >E. coli S>P. aeruginosa R (resistant) >E. coli R>B. subtilis>Z. ramigera>C. albicans>S. cerevisiae>A. pullulans B (black)>A. pullulans W (white)>T. viride Br (brown)>M. gypseum>A niger>F. nivale>P. chrysogenum>T. viride G (green).     

Antimicrobial effects of the macrocyclic Cu(II)-tetraanhydroaminobenzaldehyde complex

The Cu(II)-tetraaza macrocyclic complex exhibited antimicrobial effects on bacteria, yeasts and filamentous fungi. The highest antibacterial activity was found withB. subtilis andS. aureus, the respective IC₅₀ values being 18 and 80 μg/L and the MIC values 50 and 1000 μg/L. A concentration of 1 mg/L exerted a bacteriocide effect onS. aureus. The MIC value forB. subtilis was 250 times lower and forP. aeruginosa 10 times lower than the corresponding values for ampicillin. The Cu-complex was inactive against all tested yeasts. The strongest antifungal effect was manifested forR. nigricans, with an IC₅₀ value under 0.1 mg/L, whereas inA. alternata the IC₅₀ was 13.5 mg/L.     

Antibacterial effect of some 2,6-disubstituted 4-anilinoquinazolines

Two synthetic 2,6-disubstituted 4-anilinoquinazolines exerted a significant effect on the G⁺ bacteriaBacillus subtilis andStaphylococcus aureus. None of 12 tested derivatives influencedEscherichia coli, Proteus mirabilis andPseudomonas aeruginosa. Derivatives having the aromatic ring non-substituted or substituted by bromine, the pyrimidine ring by phenyl, morpholine or piperidine and the aniline skeleton non-substituted or substituted by methyl or amino group exterted a considerable antibacterial activity.     

Structure-activity relationships of some 4-quinazolylthiosemicarbazides and their triazolo derivatives

Eigh 4-quinazolylthiosemicarbazides and nine of their structural analogues have been tested for antibacterial effects and for structure activity relationships. 9-Chloro-5-morpholino-1,2,4-triazolo[4,3-c]quinazoline-3-thione has demonstrated the hightest antibacterial effect (MIC of 1 mg/L forE. coli andP. mirabilis and <1 mg/L forS. aureus andB. subtilis). The most effective derivatives have the carbon aromatic ring substituted with chlorine and the pyrimidine ring with morpholine or with secondary amine group.     

Immunological and biological relationship among flagellin of <Emphasis Type="Italic">Pseudomonas aeruginosa,Burkholderia cepacia</Emphasis>, and <Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis>

The flagellar protein (flagellin) was isolated and purified from strains of Pseudomonas aeruginosa, Burkholderia cepacia and Stenotrophomonas maltophilia. A significant difference was observed in the molecular weight of different flagellin preparations obtained from these bacterial isolates. Antiserum prepared against S. maltophilia flagellin did not react with flagellin of P. aeruginosa or/and B. cepacia on Immunoblot or in indirect ELISA. In addition the anti-flagellin did not agglutinate P. aeruginosa and B. cepacia. No inhibition of motility of P. aeruginosa and B. cepacia was observed in presence of antiserum; though the latter inhibited the motility of S. maltophilia. The results of the present study prove that no specific relationship existed among all the studied flagellar proteins obtained from closely related bacteria.     

EVALUATION OF COMPETITIVE ABILITY OF STAURASTRUM LUETKEMUELLERII (CHLOROPHYCEAE) AND MICROCYSTIS AERUGINOSA (CYANOPHYCEAE) UNDER P LIMITATION1

The desmid Staurastrum luetkemuellerii Donat et Ruttner and the cyanobacterium Microcystis aeruginosa Kütz. showed pronounced differences in chemical composition and ability to maintain P fluxes. The cellular P:C ratio (Q_p) and the surplus P:C ratio (Q_sp) were higher in M. aeruginosa, indicating a lower yield of biomass C per unit of P. The subsistence quota (Q_p) was 1.85 μg P·mg C^?1in S. luetkemuellerii and 6.09 μg P·mg C^?1in M. aeruginosa, whereas the respective Q_p of P saturnted organisms (Q_s) were 43 and 63 μg P·mg C^?1. These stores could support four divisions in S. luetkemuellerii and three divisions in M. aeruginosa, which suggests that the former exhibited highest storage capacity (Q_s/Q₀). M. aeruginosa showed a tenfold higher activity of alkaline phosphatase than S. luetkemuellerii when P starved. The optimum N:P ratio (by weight) was 5 in S. luetkemuellerii and 7 in M. aeruginosa. The initial uptake of P_i pulses in the organisms was not inhibited by rapid (<1 h) internal feedback mechanisms and the short term uptake rote could be expressed solely as a function of ambient P_i. The maximum cellular C-based uptake rate (V_m) in P starved M. aeruginosa was up to 50 times higher than that of S. luetkemuellerii. It decreased with increasing growth rate (P status) in the former species and remained fairly constant in the latter. The corresponding cellular P-based value (U_m= V_m/Q_p) decreased with growth rate in both species and was about 10 times higher in P started M. aeruginosa than in S. luetkemuellerii. The average half saturation constant for uptake (K_m) was equal for both species (22 μg P·L^?1) and varied with the P status. S. luetkemuellerii exhibited shifts in the uptake rate of P_i that were characterized by increased affinity (U^m/K^m) at low P_i, concentrations (<4 μg P·L^?1) compared to that at higher concentrations. The species thus was well adapted to uptake at low ambient P_i, but M. aeruginosa was superior in P_i uptake under steady state and transient conditions when the growth rate was lower than 0.75 d^?1. Moreover, M. aeruginosa was favored by pulsed addition of P_i. M. aeruginosa relpased P_i at a higher rate than S. luetkemuellerii. Leakage of P_i from the cells caused C-shaped μ vs. P_i curves. Therefore, no unique K_s for growth could be estimated. The maximum growth rate (μ_m) (23° C) was 0.94 d^?1for S. luetkemuellerii and 0.81 d^?1for M. aeruginosa. The steady state concentration of P_i (P*) was lower in M. aeruginosa than in S. luetkemuellerii at medium growth rates. The concentration of P_i at which the uptake and release of P_i was equal (P_c was, however, lower in S. luetkemuellerii.     

Thiosulfate oxidation by obligately heterotrophic bacteria

Thiosulfate was oxidized stoichiometrically to tetrathionate during growth on glucose byKlebsiella aerogenes, Bacillus globigii, B. megaterium, Pseudomonas putida, two strains each ofP. fluorescens andP. aeruginosa, and anAeromonas sp. A gram-negative, rod-shaped soil isolate, Pseudomonad H_w, converted thiosulfate to tetrathionate during growth on acetate. None of the organisms could use thiosulfate as sole energy source. The quantitative recovery of all the thiosulfate supplied to heterotrophic cultures either as tetrathionate alone or as tetrathionate and unused thiosulfate demonstrated that no oxidation to sulfate occurred with any of the strains tested. Two strains ofEscherichia coli did not oxidize thiosulfate. Thiosulfate oxidation in batch culture occurred at different stages of the growth cycle for different organisms:P. putida oxidized thiosulfate during lag and early exponential phase,K. aerogenes oxidized thiosulfate at all stages of growth, andB. megaterium andAeromonas oxidized thiosulfate during late exponential phase. The relative rates of oxidation byP. putida andK. aerogenes were apparently determined by different concentrations of thiosulfate oxidizing enzyme. Thiosulfate oxidation byP. aeruginosa grown in chemostat culture was inducible, since organisms pregrown on thiosulfate-containing media oxidized thiosulfate, but those pregrown on glucose only could not oxidize thiosulfate. Steady state growth yield ofP. aeruginosa in glucose-limited chemostat culture increased about 23% in the presence of 5–22 mM thiosulfate, with complete or partial concomitant oxidation to tetrathionate. The reasons for this stimulation are unclear. The results suggest that heterotrophic oxidation of thiosulfate to tetrathionate is widespread across several genera and may even stimulate bacterial growth in some organisms.     

Comparative in vitro activity of moxalactam (LY127935), other beta-lactam antibiotics,and aminoglycosides

Moxalactam (LY127935), a novel beta-lactam antibiotic, was compared with semisynthetic penicillins, cephalosporins, and aminoglycosides by the agar dilution method against 5,317 recent clinical isolates of facultative and anaerobic bactria. At 0.5 μg/ml, moxalactam inhibited 90% of all Gram-negative bacilli tested except forPseudomonas aeruginosa (81% inhibited by 32 μg/ml) andAcinetobacter calcoaceticus (88% inhibited by 32 μg/ml). More than 90% ofBacteroides fragilis andStaphylococcus aureus were inhibited by 4 μg/ml and 8 μg/ml, respectively. Moxalactam was at least 16-fold more active by weight than cephalothin, cefamandole, and cefoxitin forEscherichia coli, Klebsiella pneumoniae, andEnterobacter species, and 2- to 4-fold more active than cefoxitin forB. fragilis. Moxalactam was 4-fold less active than cefamandole and cephalothin forS. aureus and 2- to 4-fold less active than piperacillin forP. aeruginosa. Moxalactam was as active or more active than the aminoglycosides for all facultative Gram-negative bacilli except forP. aeruginosa. Moxalactam was inhibitory (minimal inhibitory concentration <16 μg/ml) for 20/27 gentamicin-resistant isolates and 8/13 amikacin-resistant organisms. Moxalactam’s in vitro activity against Gram-negative bacilli is markedly superior to presently available cephalosporins and, except forP. aeruginosa, is comparable to the aminoglycosides.     

Evaluation of microbial surfactants for recovery of hydrophobic pollutants from soil

Summary Several microbially produced biosurfactants were evaluated for their ability to remove hydrophobic compounds from soil. The biosurfactants produced byPseudomonas aeruginosa UG2 andAcinetobacter calcoaceticus RAG-1 displayed the best results, with recovery of [¹⁴C]hexachlorobiphenyl from soil slurries of 48.0 and 41.9%, respectively.P. aeruginosa UG2 produced higher levels of extracellular biosurfactants than four otherP. aeruginosa strains.P. aeruginosa UG2 culture filtrate containing biosurfactants enhanced the recovery of several other individual hydrocarbons and polychlorinated biphenyl compounds, as well as several hydrocarbons in a mixture, from soil. The results, suggest that biosurfactants produced byP. aeruginosa UG2 have the potential for remediation of hydrophobic pollutants in soil environments.     

Antibacterial effect of substituted 4-quinazolylhydrazines and their arylhydrazones determined by a modified microdilution method

Eight 4-quinazolylhydrazines and eleven their arylhydrazones have been tested for antibacterial effects and for structure-activity relationships by a modifed microdilution method. The derivative 6-chloro-2-morpholino-4-quinazolyl-5′-nitro-2′-furylhydrazone had the highest antibacterial effect, the MIC values being 100 mg/L forE. fœcalis, 250 mg/L forS. aureus, 200 mg/L forP. aeruginosa and 350 mg/L forE. coli. The most effective derivatives were those with the benzene ring substituted with chlorine or methyl group in position 6 or 8 and with pyrimidine ring substituted with a secondary amine in position 2. The modified microdilution method did not give rise to any statistically significant deviations in the MIC values for ampicillin in comparison with reported reference collection values. Dedicated to Professor Vladimír Betina, DSc. on the occasion of his 65th birthday     

基于16S rRNA和rpoB基因的分子系统发育分析在铜绿假单胞菌鉴定中的应用

为对比16S rRNA和rpo B基因分子系统发育分析与传统表型分类法对铜绿假单胞菌的鉴定,评估16S rRNA和rpo B基因序列分析在铜绿假单胞菌鉴定中的应用,用表型分类方法对临床自动微生物鉴定系统鉴定为铜绿假单胞菌的23株分离株进行再鉴定,PCR扩增23株分离株16S rRNA和rpo B基因片段,并测序进行系统发育分析。结果表明,表型再鉴定结果与自动微生物鉴定系统鉴定结果一致。基于两个基因的系统发育分析均显示分离株p22与不动杆菌属序列聚为一枝,其余22株分离株与铜绿假单胞菌序列聚为一枝。因此p22应鉴定为不动杆菌,16S rRNA和rpo B基因序列分析均能准确鉴定铜绿假单胞菌并能较好建立假单胞菌属内种间关系。     

An analysis of the organization and evolution of type 4 fimbrial (MePhe) subunit proteins

Summary We have analyzed and compared the amino acid sequences of the type 4 fimbrial subunits fromPseudomonas aeruginosa, Moraxella bovis, M. nonliquefaciens, Bacteroides nodosus, Neisseria gonorrhoeae, andN. meningitidis. We propose a consensus sequence for the highly conserved aminoterminal regions of these proteins. In the variable regions, a domain corresponding to an epitope common toN. gonorrhoeae andN. meningitidis fimbriae is conserved, both in sequence and in environment, in fimbrial subunits fromB. nodosus. The subunits fromM. bovis andP. aeruginosa do not show any homologies to this sequence. In all of the subunits, the carboxy-terminal half of the molecule consists of a series of fairly hydrophobic domains. The last three domains, two of which include the cysteines of the disulfide bridge inN. gonorrhoeae, P. aeruginosa, andM. bovis, are more or less conserved in sequence in all of the proteins including that ofB. nodosus. We propose that these conserved hydrophobic regions, which have the potential to form a series of beta-sheets, form a structural framework around which more variable hydrophilic sequences determining immunological profile are arranged. The evolutionary relationships of the contemporary proteins and the distribution of type 4 fimbriae are also discussed.     

Host choice and fitness correlates for conopid flies parasitising bumblebees

Two parasitoid flies,Physocephala rufipes andSicus ferrugineus (Diptera, Conopidae), and their hosts,Bombus spp., coexist at various locations in northwestern Switzerland. A detailed field study showed that both conopid species use the hostB. pascuorum to a similar degree, while the hostB. terr-luc (a pooled category ofB. terrestris andB. lucorum) is more frequently parasitised than expected byS. ferrugineus. The hostB. lapidarius in turn is exclusively used byP. rufipes. Furthermore, hosts ofB. terr-luc andB. pascuorum parasitised byS. ferrugineus were larger than hosts parasitised byP. rufipes, or than those not parasitised. The findings suggest thatS. ferrugineus selects larger hosts and may displaceP. rufipes. Pupal weight, a predictor of adult body size and parasitoid fecundity, is positively correlated with host size and larger pupae are more likely to emerge, while host species had no effect on the probability of emergence in either conopid species. Host species affected pupal weight inS. ferrugineus, but not inP. rufipes, althoughP. rufipes grew larger in hosts of a given size. Daughters emerged from larger pupae than males, but this did not correlate with larger host sizes. These observations add to the scarce knowledge of dipteran parasitoids.     

Mycoflora in cystic fibrosis: Some ecologic aspects of pseudomonas aeruginosa and Candida albicans

The mycotic and bacterial flora of 65 patients with cystic fibrosis was studied.C. albicans andP. aeruginosa were present in 33% and 43% of sputa samples, respectively; only 6.5% harbored both organisms. The mycotic flora of the nasopharynx, rectum and skin of the cystic fibrosis patients was similar to that of children with other chronic lung diseases and to that of normal children.In vitro studies clearly revealed inhibition ofC. albicans byP. aeruginosa. It is suggested thatP. aeruginosa, so prevalent with cystic fibrosis, has an inhibitory effect onC. albicans and that this interaction is effective to some extent in preventing candidal infection.Supported in part by Cancer Center Training Grant CA-08480 and Clinical Training Grant CA-08151 from the National Cancer Institute, and by ALSAC.     

Chitinase-mediated destructive antagonistic potential of Pseudomonas aeruginosa GRC1 against Sclerotinia sclerotiorum causing stem rot of peanut

Pseudomonas aeruginosa GRC₁ exhibited strong antagonistic activity against Sclerotinia sclerotiorum, in vitro and in vivo. Scanning electron microscopic (SEM) studies showed morphological abnormalities such as perforation, lysis and fragmentation of hyphae of S. sclerotiorum caused by P. aeruginosa GRC₁. This strain produced extracellular chitinase enzyme, the role of which was clearly demonstrated through Tn5 mutagenesis. Bacterization of peanut seeds with GRC₁ resulted in increased seed germination and reduced stem-rot of peanut in S. sclerotiorum-infested soil by 97%. Other vegetative and yield plant parameters such as nodules per plant, pods and grain yield per plant were enhanced with a statistical significance in comparison to control. Neomycin resistant (GRC₁^neo+) bacterium was a good root colonizer and frequently isolated from rhizosphere of peanut plants. These findings showed P. aeruginosa GRC₁ as a potential biocontrol agent against S. sclerotiorum.     

Comparative hydrocarbon utilization by hydrophobic and hydrophilic variants of Pseudomonas aeruginosa

Aims: To investigate hydrocarbon degradation by hydrophobic, hydrophilic and parental strains of Pseudomonas aeruginosa. Methods and Results: Partitioning of hydrocarbon‐degrading P. aeruginosa strain in a solvent/aqueous system yielded hydrophobic and hydrophilic fractions. Exhaustive partitioning of aqueous‐phase cells yielded the hydrophilic variants (L), while sequential fractionation of the hydrophobic phase cells yielded successive fractions exhibiting increasing cell‐surface hydrophobicity (CSH). In hydrocarbon adherence assays (bacterial attachment to hydrocarbon), L had a value of 20%, which increased from 61·7% in first hydrophobic fraction (H₁) to 72·2% in the third (H₃). Crude oil degradation by L was 70%, but increased from 82% in H₁ to 93% in H₃. L variant produced most exopolysaccharides and reduced surface tension from about 73 to 49 mN m^?1. Rhamnolipid production was highest in L, but was not detected in all crude oil cultures. Conclusions: Hydrophobic subpopulations of hydrocarbon‐degrading P. aeruginosa exhibited greater hydrocarbon‐utilizing ability than hydrophilic ones, or the parental strain. Significance and Impact of the Study: Results demonstrate that a population of P. aeruginosa consists of cells with different CSH which affect hydrocarbon utilization. This potentially provides the population with the capacity to utilize different hydrophobic substrates found in petroleum. Judicious selection of such hydrophobic subpopulations can enhance hydrocarbon pollution bioremediation.     

Culture condition ofPseudomonas aeruginosa F722 for biosurfactant production

Pseudomonas aeruginosa F722 produces a biosurfactant (BS) during its degradation of carbon and hydrocarbon compounds. The culture conditions for upgrading the biosurfactant productivity were investigated. The concentration of the biosurfactant produced byP. aeruginosa F722 was 0.78 g/L in C-medium; however, this increased to 1.66 g/L in BS medium, which was experimentally adjusted to optimal conditions. NaNO₂ was found to be most effective for microbial growth, with an O.D_600nm of 1.18 for 0.1% NaNO₂. Microbial growths, according to the O.D_600nm were 2.53, 2.68, 2.89, and 2.87 for glucose, glycerol,n-C₁₀, andn-C₂₂, respectively. Clear zone diameters (cm), indicating biosurfactant activity, were 9.0, 8.8, 5.7, and 8.5 for glucose, glycerol,n-C₁₀, andn-C₂₂, respectively. Microbial growth was not consistent with the biosurfactant activity. The best biosurfactant activity was found with a C/N ratio of 20. Under optimal culture condition, the average surface tension decreased from 70 to 30 mN/m after 5 days. With aeration of 1.0 vvm, the biosurfactant produced increased to 1.94 g/L (up to 20%) compared to that of 1.66 g/L with no aeration. With aeration, the velocities of glucose degradation during both the log and stationary growth phases increased from 0.25 and 0.18 h⁻¹ to 0.33 and 0.29 h⁻¹, respectively, and the time for the culture to arrive at the maximum clear zone diameter became shorter, from 80 down to 60 h with no aeration.     

Isolation and characterization of acetonitrile utilizing bacteria

Summary Bacteria utilizing high concentrations of acetonitrile as the sole carbon source were isolated and identified asChromobacterium sp. andPseudomonas aeruginosa. Maximum growth was attained after 96 h of incubation andP. aeruginosa grew slightly faster thanChromobacterium sp. The strains were able to grow and oxidize acetonitrile at concentrations as high as 600 mM. However, higher concentrations inhibited growth and oxygen uptake. Degradation studies with (¹⁴C)acetonitrile indicated 57% of acetonitrile was degraded byPseudomonas aeruginosa as compared to 43% byChromobacterium. The isolates utilized different nitrile compounds as carbon substrates.     

COMPETITION BETWEEN STAURASTRUM LUETKEMUELLERII (CHLOROPHYCEAE) AND MICROCYSTIS AERUGINOSA(CYANOPHYCEAE) UNDER VARYING MODES OF PHOSPHATE SUPPLY1

The desmid Staurastrum luetkemuellerii Donat et Ruttner and the cyanobacterium Microcystis aeruginosa Kütz. were grown in mixed cultures with various phosphate (P_i) additions. One pulse of P_i each day (semi-continuous cultures) favored M. aeruginosa whereas S. luetkemuellerii was favored when the same quantity of P_i was supplied continuously (chemostats). Both species coexisted under P limitation provided that the nutrient was supplied in an appropriate mode. The ability of each species to compete for P depended on their P_i uptake characteristics and their capability to retain the accumulated P_i. High affinity in uptake at low P_i concentrations contributed considerably to the growth eficiency of S. luetkemuellerii under continuous supply of P_iM. aeruginosa was, however, consistently superior to S. luetkemuellerii in accuniulatiug the newly added P, but had a high rate of P_i release. In both -types of cultures, a net high of P went from M. aeruginosa to S. luetkemuellerii. The kinetic characteristics of the two species were used to simulate the outcome of competition experiments. Simulations agreed with the experimental data f both uptake and P_i release were considered in the model. The zlariable P*(the concentration of P_i at which the net uptake is equal to μ·Q_P is a function of uptake and release of P_i but could not explain the chemostat results. S. luetkemuellerii was the winner in many experiments even if its P*was higher thou that of M. aeruginosa. Thus, in the present case P_c (the concentration at which the net uptake is zero) was a better predictor of the ability to compete for P_i under steady state as well as transient conditions in the P_i supply.