Distribution of Pseudomonas aeruginosa in a riverine ecosystem

The distribution of Pseudomonas aeruginosa in navigation pool 8 of the upper Mississippi River was investigated by acetamide broth enrichment of water, sediment, and swab (solid-water interface) samples. Among the 152 P. aeruginosa isolates, serological type 1 was most prevalent (34.2%), and a small number (13.2%) showed carbenicillin resistance. Pigmentation was variable, with only 44.7% elaborating typical blue-green pigment. P. aeruginosa was most commonly isolated from sediment, with solid-water interfaces (aufwuchs samples) also exhibiting high frequencies of isolation. Current velocity, oxygen and nutrient availability, surface tension, desiccation, and negative phototropism were important factors in the riverine distribution of this epibacterium.     

Application of DNA probes to analysis of bacteriophage distribution patterns in the environment.

Radiolabeled bacteriophage DNA probes have been used in this study to determine the distribution of Pseudomonas aeruginosa-infecting bacteriophages in natural samples of lake water, sediment, soil, and sewage. The sensitivity of detection of bacteriophage with the DNA probes was between 10(3) and 10(4) PFU and 10(6) to 10(7) CFU of lysogenized bacteria detectable with a homologous phage DNA probe. Analyses of environmental samples suggest that up to 40% of P. aeruginosa in natural ecosystems contain DNA sequences homologous to phage genomes. By using different bacteriophage DNA probes, the diversity of the bacteriophage population in sewage was estimated to be higher than that in other natural samples. The indication that transducing phages and prophages are widely distributed in the Pseudomonas populations investigated has considerable implications for the frequency of natural gene transfer by transduction and of lysogenic conversion of host bacteria in natural ecosystems.     

Application of DNA probes to analysis of bacteriophage distribution patterns in the environment.

Radiolabeled bacteriophage DNA probes have been used in this study to determine the distribution of Pseudomonas aeruginosa-infecting bacteriophages in natural samples of lake water, sediment, soil, and sewage. The sensitivity of detection of bacteriophage with the DNA probes was between 10(3) and 10(4) PFU and 10(6) to 10(7) CFU of lysogenized bacteria detectable with a homologous phage DNA probe. Analyses of environmental samples suggest that up to 40% of P. aeruginosa in natural ecosystems contain DNA sequences homologous to phage genomes. By using different bacteriophage DNA probes, the diversity of the bacteriophage population in sewage was estimated to be higher than that in other natural samples. The indication that transducing phages and prophages are widely distributed in the Pseudomonas populations investigated has considerable implications for the frequency of natural gene transfer by transduction and of lysogenic conversion of host bacteria in natural ecosystems.     

Contribution of Pseudomonas Aeruginosa strains to infections in patients of specialistic outpatients clinics of the SP ZOZ in Nidzica

The aim of this study was to examine a frequency of isolation and analysis of drug susceptibility o P. aeruginosa strains cultured from clinical specimens obtained from patients treated in specialistic outpatient clinics of the Samodzielny Publiczny Zespó? Opieki Zdrowotnej (SP ZOZ) in Nidzica durin 40 months (01. 09. 2000 - 31. 12. 2003). Ninety six P. aeruginosa strains were cultured out of 829 clinical samples collected from ambulatory patients and processed in the Bacteriological Laboratory of SP ZOZ in Nidzica during over three years. P. aeruginosa strains were isolated from 11.6% of examined specimens. The greatest number of strains (49.0%) were cultured from urine samples obtained from children. Identification of strains was performed using biochemical tests (Becton Dickinson, Emapol, bio-Merieux). Susceptibility of strains to antimicrobial agents was determined with disc diffusion method according to NCCLS recommendations. Special tests were applied to detect extended-spectrum beta-lactamases (ESBL). The most active in vitro against isolated P. aeruginosa strains was a carbapenem - imipenem. All strains were susceptible to this antibiotic. Ciprofloxacin (94.8% of susceptible strains), ceftazidime (89.6%), gentamicin (86.5%), piperacillin (84.4%) and aztreonam (76.0%) were active against the majority of P. aeruginosa strains isolated from ambulatory patients. Six strains (6.25% of all strains) producing extended--spectrum beta--lactamases (ESBL) were detected. It is alarming, that the majority of P. aeruginosa strains from outpatients were cultured out of pediatric samples (61.5%). Because of an increase in resistance and appearance of new mechanisms of resistance to antibiotics/chemotherapeutics in P. aeruginosa strains, it is necessary to monitor a drug susceptibility of these strains causing infections in ambulatory patients.     

The significance of testing for Pseudomonas aeruginosa in recreational seawater beaches.

The occurrence of Pseudomonas aeruginosa and faecal coliforms in Mediterranean sea water from beaches was investigated. Water samples (1,598 in toto) were tested and P. aeruginosa was found in 222 samples (14%). In 31% of samples where P. aeruginosa was detected, faecal coliforms of less than ten bacteria per 100 ml were found. In a group of 98 samples which had > 500 faecal coliforms per 100 ml, 41% had no detectable P. aeruginosa. The inclusion of P. aeruginosa as an additional parameter for approving beaches for recreational activity is recommended.     

Occurrence of Staphylococcus aureus and Pseudomonas aeruginosa in Israeli coastal water

The occurrence of Pseudomonas aeruginosa and coagulase-positive Staphylococcus aureus in seawater from beaches of central Israel was investigated from June 1983 until June 1985. P. aeruginosa was monitored in 652 samples of seawater from 34 beaches, and S. aureus was monitored in 628 samples. P. aeruginosa was found in 44.8% of samples (6.5% with 1 bacterium per 100 ml of water), and S. aureus was recovered from 60.7% of samples (5.3% with 1 organism per 100 ml), compared with 91.6% of samples with total coliforms (TC) and 82.2% with fecal coliforms (FC). The correlation between the presence of P. aeruginosa to that of TC and FC was 99.1 and 98.3%, respectively, while S. aureus was found in 4.3 and 8% of samples where TC and FC, respectively, were absent. Monitoring of S. aureus as a supplementary indicator in populated beaches is recommended because it will add valuable information on the sanitary quality of the seawater.     

Vertical distribution and chemical character of sediment phosphorus in two shallow estuaries in the Baltic Sea

The vertical distribution of various phosphorus (P) forms and their relation to physico-chemical properties of estuary sediment material were studied to better understand the potential release and burial of P. Core samples were taken from two dissimilar estuaries in the Baltic Sea: one in the Archipelago Sea (AS) and one in the Gulf of Finland (GoF). The P reserves were characterized by a sequential extraction procedure including the analysis of simultaneously dissolved elements in two extraction steps. The sediment material was also analysed for particle size distribution and total elements. In addition, several environmental variables were determined. The occurrence of the various forms of P varied with sediment depth among different sites. Reductant soluble, iron (Fe) bound P was the most dynamically changing P form in the sediment, while P bound to other metal oxides and apatite-P were the most stable fractions. High sedimentation rate was a dominating factor for sediment P burial. In addition, the content of organic matter, the amount of erosion-transported sorption components, and the oxygen (O₂) conditions in the near-bottom water were important determinants of the behaviour of sediment P. The results indicate that, over the long term, both estuaries have acted as sinks for deposited P and restricted the transport of P to the AS and the open GoF, thereby partly alleviating the eutrophication process.     

Occurrence of Staphylococcus aureus and Pseudomonas aeruginosa in Israeli coastal water.

The occurrence of Pseudomonas aeruginosa and coagulase-positive Staphylococcus aureus in seawater from beaches of central Israel was investigated from June 1983 until June 1985. P. aeruginosa was monitored in 652 samples of seawater from 34 beaches, and S. aureus was monitored in 628 samples. P. aeruginosa was found in 44.8% of samples (6.5% with 1 bacterium per 100 ml of water), and S. aureus was recovered from 60.7% of samples (5.3% with 1 organism per 100 ml), compared with 91.6% of samples with total coliforms (TC) and 82.2% with fecal coliforms (FC). The correlation between the presence of P. aeruginosa to that of TC and FC was 99.1 and 98.3%, respectively, while S. aureus was found in 4.3 and 8% of samples where TC and FC, respectively, were absent. Monitoring of S. aureus as a supplementary indicator in populated beaches is recommended because it will add valuable information on the sanitary quality of the seawater.     

Agricultural Plants and Soil as a Reservoir for Pseudomonas aeruginosa

Pseudomonas aeruginosa was detected in 24% of the soil samples but in only 0.13% of the vegetable samples from various agricultural areas of California. The distribution of pyocin types of soil and vegetable isolates was similar to that of clinical strains, and three of the soil isolates were resistant to carbenicillin. Pseudomonas aeruginosa multiplied in lettuce and bean under conditions of high temperature and high relative humidity (27 C and 80-95% relative humidity) but declined when the temperature and humidity were lowered (16 C, 55-75% relative humidity). The results suggest that soil is a reservior for P. aeruginosa and that the bacterium has the capacity to colonize plants during favorable conditions of temperature and moisture.     

Distribution of Plesiomonas shigelloides in various components of pond ecosystems in Dhaka, Bangladesh.

Plesiomonas shigelloides is considered to be a waterborne agent of human gastroenteritis. An ecological study was carried out in five ponds in Dhaka city over a period of one year to elucidate the distribution and seasonality of this organism in various components of pond ecosystems. Samples were collected from hydrophytes, water, phytoplankton and sediment every 15 days over 12 months and cultured for P. shigelloides. P. shigelloides was isolated from a total of 120 samples including 25 (20.8%), 16 (13.3%), 22 (18.3%) and 35 (29.2%) of hydrophytes, water, phytoplankton and sediment samples, respectively. Distinct seasonal patterns of isolation of P. shigelloides were observed in the four components with two distinct peaks. The highest peaks were observed in hydrophytes and water samples in May and in phytoplankton and sediment in November. P. shigelloides was isolated from all components from all ponds during the study period. These results suggest that P. shigelloides is an autochthonous member in the freshwater pond ecosystems in Dhaka, Bangladesh.     

Immunological study of the cellular components of Pseudomonas aeruginosa. VI. The toxicity and protective properties of its mucus and its fractions]

Capsule-like mucus was obtained from two newly isolated mucoid strains of P. aeruginosa and then treated with ethanol. The mucus was fractionated by the method of differential centrifugation (at 15, 000 g for 1 h, at 105, 000 or 170, 000 g for 3 h) and gel chromatography in columns packed with Sepharose 4B. The sediment fractions contained 30--80% of high molecular polysaccharide protein (peptide) mucus components which were toxic for mice and protected 25--77% of rats against experimental P. aeruginosa infection. The supernatant fluid fractions contained 60--80% of predominantly protein components with molecular mass between 20,000 and 60,000 daltons. These mucus components were slightly toxic for mice and rats and protected 80--100% of rats against P. aeruginosa infection.     

Routes of the transmission of Pseudomonas aeruginosa infection in resuscitation and intensive therapy departments

To find out if the transfer of P. aeruginosa infection by droplet route is possible in resuscitation and intensive care units, the bacteriological study of air samples taken in different rooms of resuscitation units (altogether 234 air samples) was carried out with the subsequent identification and typing of isolated P. aeruginosa strains. In most cases (70.5%) the microbial contamination of the air in the main rooms of resuscitation units was found not to exceed 500 microbial cells per cu. m, and no P. aeruginosa strains were isolated. The identification and typing of six P. aeruginosa strains isolated from the air of an isolation ward for patients with infectious complications made it possible to find out intraspecific differences of these microorganisms, as all of them belonged to strains of different sero- and pyocinotypes. Thus, the results of these investigations indicate that the droplet route of the transfer of P. aeruginosa hospital infection is not characteristic of resuscitation and intensive care units, as no P. aeruginosa strains are isolated from the main rooms of such units; likewise, no circulation of this microorganism was observed in the air of an isolation ward for patients with infectious complications.     

Aerobic Bacterial Flora of Semen and Stallion Reproductive Tract and its Relation to Fertility Under Field Conditions

This study was initiated in order to investigate the bacterial flora of the stallion genital tract by taking consecutive samples from normal stallions in regular use. The objective was to determine whether any growth of potential pathogens, particularly P. aeruginosa and K. pneumoniae, in fresh semen and urethra was associated with the presence of inflammatory cells in the semen and whether bacterial growth had any effect on sperm morphology and pregnancy results. Sixteen stallions, only used for A.I., housed at 3 different commercial stud farms, were used. A wide variety of microorganisms was found in almost all samples from fresh semen (total 115 samples). P. aeruginosa was isolated from 46/115 (40%) of the samples and from 12 of the 16 stallions. K. pneumoniae was isolated from the semen of one stallion. Samples taken from the distal urethra after ejaculation contained fewer microorganisms than samples from fresh semen. No bacteria were found in 51% of the extended semen samples. Most of the stallions had an acceptable sperm morphology, and very few of the ejaculates contained inflammatory cells. Pregnancy results among the stallions varied, but were acceptable for most of them. There was no correlation between the frequency of samples testing positive for P. aeruginosa in raw semen and pregnancy results. kw|Keywords|k]bacterial growth k]urethra k]Pseudomonas aeruginosa k]Klebsiella pneumoniae     

Distribution of microorganisms lysing Pseudomonas aeruginosa and Staphylococcus aureus in the system of sewage waters

The regularities of lytic microorganisms distribution in domestic sewage have been studied. A reproduction of mesophilic gram-negative bacteria producing lytic substances against Pseudomonas aeruginosa and Staphylococcus aureus has been shown to take place at mechanical cleaning stages. In the primary sediment trap the number and the relative content of microorganisms lysing P. aeruginosa at mean temperature and the number of microorganisms lysing S. aureus are maximum. The number of gram-positive sporogenous bacteria lysing P. aeruginosa under conditions close to thermophilic does not change considerably till the secondary sediment trap and remains comparatively high. Certain stages of purification can be regarded as a source of microorganisms producing lytic substances.     

A protease stable in organic solvents from solvent tolerant strain of Pseudomonas aeruginosa

A solvent tolerant strain of Pseudomonas aeruginosa (PseA) was isolated from soil samples by cyclohexane enrichment in medium. The strain was able to sustain and grow in a wide range of organic solvents. The adaptation of P. aeruginosa cell towards solvents was seen at membrane level in transmission electron micrographs. It also secreted a novel protease, which exhibited remarkable solvent stability and retained most of the activity at least up to 10 days in the presence of hydrophobic organic solvents (log P > or = 2.0) at 25% (v/v) concentrations. The protease was able to withstand as high as 75% concentration of solvents at least up to 48 h. P. aeruginosa strain and its protease, both seem promising for solvent bioremediation, wastewater treatment and carrying out biotransformation in non-aqueous medium.     

Characterization of pseudomonads isolated from diseased fleece.

A total of 59 Pseudomonas isolates was obtained from 11 samples of diseased fleece taken from live sheep. All but four of the isolates could be assigned to one of nine Pseudomonas species, of which P. aeruginosa, P. alcaligenes, P. mendocina , and P. putida were the most common. P. aeruginosa was found in four of the fleece samples and, when present, appeared to predominate. Although several of the isolates of P. aeruginosa lacked the ability to produce pyocyanin (and some produced neither pyocyanin nor fluorescein), nearly all produced several virulence factors. Of the other pseudomonads, many produced proteinase, esterase, and catalase, several were able to grow at 42 degrees C and reduce nitrate, and some also produced lipase and hemolysin and, like P. aeruginosa, might serve to initiate (or sustain) the dermatitis frequently associated with fleece rot in sheep.     

Characterization of pseudomonads isolated from diseased fleece

A total of 59 Pseudomonas isolates was obtained from 11 samples of diseased fleece taken from live sheep. All but four of the isolates could be assigned to one of nine Pseudomonas species, of which P. aeruginosa, P. alcaligenes, P. mendocina , and P. putida were the most common. P. aeruginosa was found in four of the fleece samples and, when present, appeared to predominate. Although several of the isolates of P. aeruginosa lacked the ability to produce pyocyanin (and some produced neither pyocyanin nor fluorescein), nearly all produced several virulence factors. Of the other pseudomonads, many produced proteinase, esterase, and catalase, several were able to grow at 42 degrees C and reduce nitrate, and some also produced lipase and hemolysin and, like P. aeruginosa, might serve to initiate (or sustain) the dermatitis frequently associated with fleece rot in sheep.     

In situ growth rates and biofilm development of Pseudomonas aeruginosa populations in chronic lung infections

The growth dynamics of bacterial pathogens within infected hosts are a fundamental but poorly understood feature of most infections. We have focused on the in situ distribution and growth characteristics of two prevailing and transmissible Pseudomonas aeruginosa clones that have caused chronic lung infections in cystic fibrosis (CF) patients for more than 20 years. We used fluorescence in situ hybridization (FISH) directly on sputum specimens to examine the spatial distribution of the infecting P. aeruginosa cells. Mucoid variants were present in sputum as cell clusters surrounded by an extracellular matrix, whereas nonmucoid variants were present mainly as dispersed cells. To obtain estimates of the growth rates of P. aeruginosa in CF lungs, we used quantitative FISH to indirectly measure growth rates of bacteria in sputum samples (reflecting the in vivo lung conditions). The concentration of rRNA in bacteria isolated from sputa was measured and correlated with the rRNA contents of the same bacteria growing in vitro at defined rates. The results showed that most cells were actively growing with doubling times of between 100 and 200 min, with some growing even faster. Only a small stationary-phase subpopulation seemed to be present in sputa. This was found for both mucoid and nonmucoid variants despite their different organizations in sputum. The results suggest that the bacterial population may be confronted with selection forces that favor optimized growth activities. This scenario constitutes a new perspective on the adaptation and evolution of P. aeruginosa during chronic infections in CF patients in particular and on long-term infections in general.     

Factors influencing the spatio-temporal distribution of benthic Microcystis aeruginosa colonies (Cyanobacteria) in the hypertrophic Grangent Reservoir (Loire, France)

The spatio-temporal distribution of benthic colonies of Microcystis aeruginosa in Grangent Reservoir (France) in 2000 was not homogeneous and appeared to be controlled by many external factors: lake depth, station morphometry, substratum and hydraulic regime (lacustrine or fluvial). A most important concentration of benthic colonies was found at deep sites with fine sediment or at sites where the sediment was rich in organic matter. In spite of a stable water level and a minimum flow during summer, the number of benthic colonies showed great variation in the lacustrine downstream part of the reservoir. These variations may be explained by the dynamics of planktonic cyanobacteria.     

Detection of Pseudomonas aeruginosa from clinical and environmental samples by amplification of the exotoxin A gene using PCR.

PCR was used to detect Pseudomonas aeruginosa from water samples by amplifying a 396-bp region of the exotoxin A (ETA) structural gene sequence. The identify of the amplified 396-bp fragment was confirmed by digesting it with PvuI restriction endonuclease, which produced the predicted 246- and 150-bp fragments. Specific primers amplified ETA-positive P. aeruginosa DNA, whereas other species of Pseudomonas and GC-rich bacteria did not yield any 396-bp fragment. The specificity and sensitivity of the assay were 100 and 96%, respectively, which confirms the assay's reliability for diagnostic and epidemiological studies. The assay can detect as few as 5 to 10 cells in a 10-ml water sample or 0.1 pg of P. aeruginosa DNA per reaction mixture (5 microliters) by ethidium bromide staining of an agarose gel. Ten-times-lower concentrations were detected by hybridization with a digoxigenin-labeled oligonucleotide probe internal to the PCR product. With this PCR method, ETA-positive P. aeruginosa was detected in animal cage water samples at a level of 40 cells per ml. This method is rapid and less cumbersome than other diagnostic methods for the identification of P. aeruginosa strains. The method described can be used to detect a low level of P. aeruginosa from environmental and clinical samples without the use of selective media or additional biochemical tests.