Detection of Pseudomonas aeruginosa

100 ml or 250 ml of the water to be analyzed is mixed with double-strength malachite-green broth and incubated for up to 44 4 hours at 37 1 C. The subcultures are then placed on a suitable selective culture medium or on endoagar and the colonies formed after an incubation time of 20 4 hours at 37 C are analyzed in terms of their ability to form fluorescin and pyocyanin. For this, the suspect colonies are inoculated onto culture mediums A and B (after King) and incubated for 44 4 hours at 37 °C. In addition, a check should be carried out as to whether ammonia is formed from acetamide. For this purpose a solution of ammonium-free acetamide is injected and checked with Nessler s reagent for the presence of ammonia after an incubation time of 20 4 hours at 37 C. 

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A displacement assay was also used for the detection of Pseudomonas aeruginosa by Bovenizer et al. [90]. However, a poor detection hmit of... 

Khan A.A. and Cemiglia C.E. (1994) Detection of Pseudomonas aeruginosa from clinical and environmental samples by amplification of the exotoxin A gene using PCR. Appl. Environ. Microbiol., 60, 3739-3745. 

Fig. 11.—Chemical structure of the two preponderant lipid A forms of Pseudomonas aeruginosa. (A) Pentaacyl lipid A (major lipid A fraction 75%, w/w). (B) Hexaacyl lipid A (minor lipid A fraction, 25%, w/w). Dashed lines indicate nonstoichiometric a-hydroxylation of 12 0. A lipid A species having 2 mol 12 0(2-OH)/mol lipid A was not detected (77).

The ferrocyanide complex is not easily biodegradable (Belly and Goodhue 1976 Pettet and Mills 1954). However, when an aqueous solution of potassium ferrocyanide was seeded with pure culture of Pseudomona aeruginosa, E. coli, or a mixture of the two bacteria, formation of free cyanide was observed after a delay period of 2 days (Cherryholmes et al. 1985). The rate of free cyanide formation increased with addition of nutrient in water, and a free cyanide concentration <4,000 pg/L was detected at the end of 25 days. It was shown that the free cyanide formation was due to biodegradation and not to either photolysis or hydrolysis. The relevance of this study to the fate of ferrocyanide complexes in natural water or industrial effluents is difficult to assess because cyanide concentrations used in these experiments (3,300 mg/L) are rarely encountered in these media. 

Rapid and sensitive HPLC methods were developed for the detection of an antimicrobial growth promoter and its main metabolites containing quinoxaline-2-carboxylic acid <2005MI1495>. The major phenazine pigments of Pseudomonas aeruginosa such as 1-hydroxyphenazine and phenazine-l-carboxylic acid <1997JCH(A)(771)99>, and 2-aminophenazine as an impurity in a bactericide <1999MI632>, were also analyzed by HPLC methods. 

The first report of a specific screening technique designed to search for p-lactam antibiotic-producing cultures was described by Kitano et al. (1975). A mutant of Pseudomonas aeruginosa highly and specifically sensitive to p-lactam antibiotics was isolated. A similar mutant strain of Escherichia coli highly sensitive to p-lactam antibiotics was used in the detection of nocardicins (Aoki et al., 1976). 

Rhamnolipid 1 and rhamnolipid 3 are the major rhamnolipids produced by using resting cells of Pseudomonas aeruginosa DSM 2874. Two further rhamnolipids that are similar in structure but contain only one hydroxydecanoic acid unit, rhamnolipids 2 and 4, have also been detected. The rhamnolipid production with resting cells is a two-step process. In a first step Pseudomonas aeruginosa cells are produced and harvested. In a second step this biomass is used for the rhamnolipid production under growth-limiting conditions [29],... 

Nucleotidylation - the addition of adenylate-residues by Lnu enzymes - can also be the cause of resistance to lincosamide antibiotics in staphylococci and enterococci. A plasmid encoded ADP-ribosylating transferase (Arr-2) that leads to rifampicin resistance has been detected in various Enterobacteriaceae as well as in Pseudomonas aeruginosa. 

Nakamura, A. Miyakozawa, L Nakazawa, K. O Hara, K. Sawai, T. Detection and characterization of macrolide 2 -phosphotransferase from a Pseudomonas aeruginosa clinical isolate. Antimicrob. Agents Chemother., 44, 3241-3242 (2000)... 

Chakrabarty et al. (1989) selected several antihistamines for detection of antibacterial action [15]. According to them, promethazine was the most powerful and significant antimicrobial. The authors observed that in staphylococci, shigellae, and vibrios the MIC of promethazine varied between 100 and 200 xg/ml. Although one strain of Escherichia coli was sensitive at 50 xg/ml, the others were resistant. With respect to Pseudomonas aeruginosa, Proteus vulgaris, and Providencia spp., the MIC of promethazine was always >200 pg/ml (Table 7). 

Fig.3. 800-MHz NMR spectra of oxidized (A) Pseudomonas aeruginosa azurin, (B) spinach plastocyanin, and (C) cucumber stellacyanin recorded in D2O solution. The letters identify the resonance of the equivalent proton in the three proteins. In the insets the far-downheld regions containing signals not observable in direct detection are shown (Bertini et al., 2000). The positions and the linewidths of the signals of the oxidized species were obtained using saturation transfer experiments over the far-downheld region by measuring the intensity of the exchange connectivity with the corresponding signal in the reduced species (Bertini et al., 1999, 2000).

A few reports on metal ion characterization in metalloenzymes by hyphenated techniques with ICP-MS detection should also be mentioned. Inhibition, reactivation, and determination of metal ions in membrane metalloproteases of bacterial origin was studied by Leopold et al The results obtained by SEC-ICP-MS and the results of enzymological methods indicated that two different membrane proteases from Bacillus cereus and Pseudomonas aeruginosa were zinc metalloproteases. In another study, ICP-MS measurements helped in the characterization of a fibrinolytic metalloprotease from the ftniting bodies of an edible mushroom. " Using HPLC-ICP-MS, Suzuki et al. detected a zinc-binding protein present specifically in the livers of male adult rats. ... 

The use of Mini-tip Culturette (Becton Dickinson, Cockeysville, MD) has been compared with traditional culture techniques using a rabbit model as well as commimity-acquired presumed bacterial keratitis. The sensitivity of the Mini-tip Culturette was 83-3% and the specificity 100%. Detected organisms included group A P-hemolytic Streptococcus, S. aureus, coagulase-negative Staphylococcus, Serratia marcescens, and Pseudomonas aeruginosa. 

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