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Degradation and mineralization of atrazine by a soil bacterial isolate.

An atrazine-degrading bacterial culture was isolated from an agricultural soil previously impacted by herbicide spills. The organism was capable of using atrazine under aerobic conditions as the sole source of C and N. Cyanuric acid could replace atrazine as the sole source of N, indicating that the organism was capable of ring cleavage. Ring cleavage was confirmed in 14CO2 evolution experiments with [U-14C-ring]atrazine. Between 40 and 50% of ring-14C was mineralized to 14CO2. [14C]biuret and [14C]urea were detected in spent culture media. Cellular assimilation of 14C was negligible, in keeping with the fully oxidized valence of the ring carbon. Chloride release was stoichiometric. The formation of ammonium during atrazine degradation was below the stoichiometric amount, suggesting a deficit due to cellular assimilation and metabolite-N accumulation. With excess glucose and with atrazine as the sole N source, free ammonium was not detected, suggesting assimilation into biomass. The organism degraded atrazine anaerobically in media which contained (i) atrazine only, (ii) atrazine and glucose, and (iii) atrazine, glucose, and nitrate. To date, this is the first report of a pure bacterial isolate with the ability to cleave the s-triazine ring structure of atrazine. It was also concluded that this bacterium was capable of dealkylation, dechlorination, and deamination in addition to ring cleavage.

Atrazine↗

A comparative study on the application of 3 molecular methods in epidemiological typing of bacterial isolates using MRSA as a prototype.

OBJECTIVE: To evaluate and compare 3 widely used molecular techniques, namely, restriction endonuclease analysis of plasmid deoxyribonucleic acid (REAP), randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) for their suitability and usefulness in the typing and fingerprinting of bacterial isolates. METHODS: Twenty-four epidemiologically unrelated methicillin-resistant Staphylococcus aureus (MRSA) isolates were used to evaluate the molecular typing methods (REAP, RAPD and PFGE). The study was conducted at the Research and Diagnostic Laboratories of King Faisal Specialist Hospital and Research Center from January 2002 through January 2003. RESULTS: Only 20.8% of all isolates studied were of the same genotypes by all 3 methods. Two major clusters of strains each representing 33% of the total number of isolates were identified by REAP analysis. Each of RAPD and PFGE however, identified one major cluster represented by 54% and 83% of the total number of isolates, all 3 typing methods, therefore, showed the clonal genetic relatedness among distant MRSA isolates. However inter-strain comparison of fingerprint data generated from each method revealed differences in clonal representation of the MRSA isolates. CONCLUSION: Although a variety of molecular assays are available for typing of bacterial species, there is no single standardized protocol for routine analysis. Reproducibility and interpretation of genotypic data are therefore, highly dependent on methodologies employed by the individual laboratory. Our findings illustrate the importance of using a combination of methods in typing schemes of bacterial isolates. In terms of reproducibility and typeability we found that PFGE is superior to REAP and RAPD and, therefore, more suitable for routine, standardized tracing of nosocomial bacterial isolates.

Anti-Bacterial Agents↗

Comparison of in vitro susceptibilities of ocular bacterial isolates to gatifloxacin and other topical antibiotics.

PURPOSE: To compare the in vitro activity of gatifloxacin with that of other topical antibiotics against fresh bacterial isolates from various types of human ocular infections. METHODS: A total of 498 strains of ocular bacterial isolates were tested for their susceptibility to gatifloxacin, ofloxacin, ciprofloxacin, norfloxacin, gentamicin, tobramycin, neomycin, chloramphenicol, erythromycin, tetracycline and amikacin. RESULTS: Ninety-nine percent of gram-positive and 92% of gram-negative bacterial strains were found to be susceptible in vitro to gatifloxacin, whereas lower percentages of gram-positive and gram-negative bacteria were found to be susceptible to other antibiotics. CONCLUSIONS: In vitro, gatifloxacin exhibits good activity against various bacterial species isolated from different ocular sites. This requires corroboration in the clinical setting.

Anti-Infective Agents, Local↗

Invasive potential of bacterial isolates associated with subclinical bovine mastitis.

This work describes differences in the invasive ability of bacterial isolates associated with mastitis. Invasion ability was determined by the uptake and survival in a primary culture of bovine mammary epithelial cells (BMEC). BMEC were isolated from a healthy lactating cow and characterized by their morphology, immunostaining for cytokeratin and the detection of beta- and kappa-casein mRNAs. Ten bacterial isolates comprising the staphylococcal species Staphylococcus aureus (3), Staphylococcus epidermidis (1), Staphylococcus haemolyticus (1), Staphylococcus equorum (2), Staphylococcus xylosus (1) and Brevibacterium stationis (2) obtained from raw milk of cows with mastitis from backyard farms were assayed for their ability to invade BMEC. Only two S. aureus and one S. epidermidis isolates were able to invade BMEC, at similar levels to the S. aureus control strain ATCC 27543. In conclusion, using the in vitro model of infection used in this study, differences in bacterial invasion capability may be detected.

Animals↗

Changing bacterial isolates and antibiotic sensitivities of purulent dacryocystitis.

OBJECTIVE: To examine the current spectrum of bacterial isolates in cases of chronic dacryocystitis and dacryoabcess and to determine their antibiotic sensitivities. MATERIALS AND METHODS: We carried out a prospective study in which 39 patients who presented to the outpatient clinic with chronic purulent dacryocystitis or acute dacryoabscess had cultures taken. All patients were antibiotic-free for at least one week prior to culturing. RESULTS: The 39 positive cultures grew 41 bacterial isolates, as two patients grew two species of bacteria; 16 isolates (39%) were Gram-positive and 25 (61%) were Gram-negative. The most common isolates were Pseudomonas (22%), Staphylococcus aureus (13%), Enterobacter (10%), Citrobacter (10%), Streptococcus pneumoniae, E. coli, and Enterococcus (7%). Uncommon Gram-negative bacteria were also cultured: Alcaligenes in two cases (5%), and one case of Stenotrophomonas maltophilia (2.5%). No anaerobic bacteria were isolated. Gram-negative isolates were sensitive to ceftazidime in 95%, ciproxin in 86%, and cefuroxime in 50% of cases, with sensitivity of less than 30% to cephalexin and ampicillin in those tested. All Pseudomonas isolates (100%) were sensitive to ceftazidine, 86% were sensitive to ciprofloxacin, with only 20% being sensitive to ampicillin and 14% to cephalexin. Alcaligenes was resistant to all antibiotics tested with the exception of ceftazidine. CONCLUSION: This study demonstrates a significant change in bacterial flora and antibiotic treatment requirements of purulent dacryocystitis from previously published data. A higher incidence of Gram-negative organisms, particularly Pseudomonas, with resistance to commonly used antibiotics was found. The emergence of rarer highly resistant Gram-negative microorganisms may also indicate a trend in lacrimal sac infections. These findings suggest that the antibiotic treatment protocol before and after lacrimal surgery should be reconsidered in this subgroup of patients.

Adolescent↗

Changing bacterial isolates and antibiotic sensitivities of purulent dacryocystitis.

PURPOSE: To examine the current spectrum of bacterial isolates in cases of chronic dacryocystitis and dacryoabcess and to determine their antibiotic sensitivities. MATERIALS AND METHODS: We carried out a prospective study in which 39 patients who presented to the outpatient clinic with chronic purulent dacryocystitis or acute dacryoabscess had cultures taken. All patients were antibiotic-free for at least one week prior to culturing. RESULTS: The 39 positive cultures grew 41 bacterial isolates, as two patients grew two species of bacteria. Sixteen isolates (39%) were Gram positive and 25 (61%) were Gram negative. The most common isolates were Pseudomonas (22%), Staphylococcus aureus (13%), Enterobacter (10%), Citrobacter (10%), Streptococcus pneumoniae, Escherichia coli, and Enterococcus (7%). Uncommon Gram-negative bacteria were also cultured: Alcaligenes in 2 cases (5%) and one case of Stenotrophomonas maltophilia (2.5%). No anaerobic bacteria were isolated. Gram-negative isolates were sensitive to ceftazidime in 95%, ciprofloxacin in 86% and cefuroxime in 50%, with a sensitivity of less than 30% to cefalexin and ampicillin in those tested. All Pseudomonas isolates (100%) were sensitive to ceftazidime, 86% were sensitive to ciprofloxacin, while only 20% were sensitive to ampicillin and 14% to cefalexin. Alcaligenes was resistant to all antibiotics tested with the exception of ceftazidime. CONCLUSION: This study demonstrates a significant change in bacterial flora and antibiotic treatment requirements of purulent dacryocystitis from previously published data. A higher incidence of Gram-negative organisms, particularly Pseudomonas, with resistance to commonly used antibiotics was found. The emergence of rarer, highly resistant, Gram-negative microorganisms may also indicate a trend in lacrimal sac infections. These findings suggest that the antibiotic treatment protocol before and after lacrimal surgery should be reconsidered in this subgroup of patients.

Adolescent↗

Production of exopolysaccharides by Antarctic marine bacterial isolates.

AIMS: This study was undertaken to examine and characterize Antarctic marine bacterial isolates and the exopolysaccharides (EPS) they produce in laboratory culture. METHODS AND RESULTS: Two EPS-producing bacterial strains CAM025 and CAM036 were isolated from particulate material sampled from seawater and sea ice in the southern ocean. Analyses of 16S rDNA sequences placed these isolates in the genus Pseudoalteromonas. In batch culture, both strains produced EPS. The yield of EPS produced by CAM025 was 30-fold higher at -2 and 10 degrees C than at 20 degrees C. Crude chemical analyses showed that these EPS were composed primarily of neutral sugars and uronic acids with sulphates. Gas chromatographic analysis of monosaccharides confirmed these gross compositional findings and molar ratios of monosaccharides revealed differences between the two EPS. CONCLUSIONS: The EPS produced by Antarctic bacterial isolates examined in this study appeared to be polyanionic and, therefore, 'sticky' with respect to cations such as trace metals. SIGNIFICANCE AND IMPACT OF THE STUDY: As the availability of iron as a trace metal is of critical importance in the southern ocean where it is know to limit primary production, the role of these bacterial EPS in the Antarctic marine environment has important ecological implications.

Antarctic Regions↗

Biotyping of bacterial isolates using the yeast killer system.

Forty-four presumptive killer yeasts were tested against bacterial isolates, including rapid-growing gram-positive and gram-negative bacteria, as well as slow-growing bacteria, such as the mycobacteria. A killer system, based on the patterns of bacterial susceptibility to the action of nine selected killer yeasts, was developed for epidemiological purposes. The killer system, previously standardized for yeasts and hyphomycetes, was adapted to the specific growth conditions of the bacterial isolates. The results obtained confirm that susceptibility to the yeast killer phenomenon is widespread among microorganisms unrelated to yeasts and that it could form the basis for a convenient and adaptable biotyping method in microbiological laboratories.

Bacterial Typing Techniques↗

Bacterial isolates in the sick young infant in Ile-Ife, Nigeria.

This prospective study was carried out to determine the bacterial pathogens and their antibiotic sensitivity profile in the sick young infant. All consecutive young infants with features suggestive of infection seen over 5 months at a Comprehensive Health Centre in Ile-Ife, were screened for septicaemia and local bacterial infections. Of the 121 sick young infants screened for infection, 94 (77.7 per cent) had confirmed bacterial infection and 54 (57.4 per cent) of the 94 had confirmed septicaemia. Gram-positive organisms were the commonest bacterial isolates accounting for 204 (81.6 per cent) of the 250 isolates in this study. Staphylococcus aureus was the most frequent organism accounting for 61.2 per cent of all isolates. Gram-negative organisms accounted for 46 (18.4 per cent) of all isolates with Salmonella spp. and Proteus vulgaris predominating. All the bacterial isolates in this study were sensitive to ofloxacin and most were sensitive to the antibiotics commonly employed in the treatment of infections caused by these organisms. However, many of both Grampositive and Gram-negative isolates in this study were resistant to cotrimoxazole. The study highlights the high prevalence of bacterial infections (localized or systemic) among young infants. It also shows that Gram-positive organisms, the principal aetiologic agents, were sensitive to commonly used antibiotics. It is recommended that genticin and cloxacillin or erythromycin should be used as the first-line antibiotics in the treatment of young infants with bacterial infections in Ile-Ife, Nigeria.

Age Distribution↗

The prevalence and antibiotic susceptibility pattern of gram-negative bacterial isolates in two ICUs in Saudi Arabia and Kuwait.

In surveys of the prevalence and antibiotic susceptibility pattern of consecutive Gram-negative bacterial isolates in two intensive care units (ICUs) in Saudi Arabia (Jeddah) and Kuwait, 106 and 101 isolates, respectively, were analyzed. The most common bacterial isolates in Jeddah versus Kuwait ICUs were Pseudomonas aeruginosa (26%, 26%), Escherichia coli (23%, 3%), Klebsiella pneumoniae (20%, 17%), inducible Enterobacteraecae group (17%, 14%), and Acinetobacter spp. (9%, 33%). Overall, about 99% of all isolates were susceptible to ciprofloxacin in both centers, whereas 87 and 96% were susceptible to imipenem, 69 and 64% to ceftazidime, 59 and 52% to cefotaxime, and 25 and 67% to piperacillin, respectively, in Jeddah and Kuwait. Prior antibiotic usage was more common among patients in Jeddah than in Kuwait. Dominant features of the study in Jeddah were the E. coli and Klebsiella spp. demonstrating multiresistance to monobactams, cephems, and all three aminoglycosides, and evidence of two classes of resistance to beta-lactam antibiotics which were not seen among the Kuwaiti isolates. The Kuwaiti Pseudomonas spp. were more sensitive to imipenem than the Jeddah Pseudomonas spp. (100% versus 68%). The higher number of resistant bacteria seen in Jeddah than Kuwaiti may be a reflection of the higher antibiotic consumption, in particular higher usage of broad spectrum cephalosporins in Jeddah ICU.

Adolescent↗

Identification and characterization of bacterial isolates from the Mir space station.

Twenty bacterial isolates (supplied by NASA) from the Mir space station water system were identified using Vitek GNI+ test card, API 20NE, and 16S rRNA gene sequencing. The identification of only one isolate agreed among the three techniques. The utility of the API 20NE and Vitek GNI+ test card approaches for identifying these isolates was Limited. Although 16S rRNA gene sequencing effectively identified many of the bacteria to the genus level, 74% of the isolates could not be identified to the species level. Isolates were also characterized based on motility and hydrophobicity. About 40% of the isolates were motile and four isolates were hydrophobic, suggesting that many of the bacteria have the potential to colonize surfaces and form biofilms. These findings demonstrate the difficulties in identifying bacteria from some environments to the species level and have implications for determining the risks of contamination in water systems of space shuttles and stations.

Bacterial Typing Techniques↗

Susceptibility of bacterial isolates to cefepime in comparison to other broad spectrum antimicrobial agents at a tertiary care center in Lebanon.

Newly introduced antimicrobial agents have to be evaluated to establish their current activity and susceptibility data base against microorganisms for future comparison. Cefepime is a fourth generation cephalosporin that was recently introduced in Lebanon but no background susceptibility data is available for it in this country. We prospectively analyzed the antimicrobial susceptibility pattern of the bacterial isolates from the American University of Beirut Medical Center to a number of broad spectrum antimicrobial agents and compared it to the susceptibility of cefepime. Consecutive clinical bacterial isolates, representing 665 gram-negative and 387 gram-positive were tested: 82 to 100% of the gram-negative isolates were susceptible to cefepime, including most of the extended spectrum beta-lactamase producing Enterobacteriaceae. All the oxacillin susceptible staphylococcus isolates, and the penicillin susceptible Streptococcus spp. as well as 92% of the S. pneumoniae isolates were susceptible. The data shows that currently, cefepime provides a very broad spectrum of activity against gram-positive as well as gram-negative bacterial isolates recovered from clinical specimens.

Cefepime↗

Fortimicin A: collaborative in vitro susceptibility. Comparison with amikacin and gentamicin against 11,840 clinical bacterial isolates.

The susceptibility of 11,840 clinical bacterial isolates to fortimicin A was determined by agar dilution or broth microdilution methods and compared with their susceptibility to amikacin and gentamicin. In general, the in vitro activity of fortimicin A was essentially the same as that of amikacin. Significant exceptions were the increased effectiveness of fortimicin A against Serratia marcescens and the greater activity of amikacin against Pseudomonas and other nonfermentative gram-negative bacilli. On a weight-for-weight basis, gentamicin showed greater activity than the other two antimicrobial drugs against most species; S. Marcescens was the major exception. However, at concentrations equivalent to achievable nontoxic serum levels, the proportion of isolates inhibited by the three drugs was quite comparable. There were several strains with unusually high resistance to one or more of the tested antibiotics. These usually occurred in one of the six participating institutions and could be traced to specific enzyme-producing or permeability mutants endemic to that particular institution.

Amikacin↗

An investigation of the in vitro anti-bacterial activity of noxythiolin against 1,000 pathogenic bacterial isolates.

The susceptibility of 1,000 recent bacterial isolates to noxythiolin was determined by the disc susceptibility method. No Gram-positive strains were resistant to this method but 56 (5.6%) of Gram-negative strains gave zones of inhibition of 12 mm diameter or less. The minimum inhibitory concentration (MIC) of the latter were determined by the agar incorporation method. No strains had MIC values greater than 4096 mg/litre. Since concentrations of 50,000 mg/litre can be used for topical treatment, these organisms may be considered susceptible.

Bacteria↗

Susceptibility patterns of bacterial isolates from intensive care and haematology/oncology patients in New Zealand.

AIMS: To determine the current susceptibility pattern of bacterial isolates from intensive care and haematology/oncology patients in New Zealand. METHOD: Over a 6 month period 417 consecutive clinically relevant bacterial isolates from intensive care and haematology/oncology patients from seven New Zealand hospitals had their susceptibility to multiple antimicrobial agents determined by the agar plate dilution method. Methicillin resistant staphylococci were not included. RESULTS: Of the 417 isolates, 224 (54%) were gram negative and 193 were gram positive. Predominant species/groups were: Escherichia coli 63 (15%), Enterobacter spp 26 (6%), other Enterobacteriacae 41 (10%), Pseudomonas aeruginosa 42 (10%), Staphylococcus aureus 111 (27%), coagulase negative staphylococci 30 (7%), Streptococcus spp 31 (7%), and Enterococcus spp 19 (5%). Isolate sources were: respiratory tract, 170 (41%); cutaneous sites, 81 (19%); blood, 64 (15%); and urine 63 (15%). Resistance was uncommon amongst staphylococci, streptococci, enterococci, and H influenzae. No vancomycin resistant or beta-lactamase-positive enterococci were encountered. For different groups of enteric gram negative bacilli: amoxycillin and amoxycillin-clavulanic acid resistance was common, 46-93% and 24-85% respectively; cefpirome was the most active cephalosporin; aminoglycoside resistance was uncommon; and no isolate possessed extended spectrum beta-lactamase. For P aeruginosa: most isolates were susceptible to cefpirome and ceftazidime, and aminoglycoside resistance was uncommon. CONCLUSION: Gram positive bacteria make up a higher proportion of isolates than in a similar European study. At present New Zealand does not have widespread resistance amongst common isolates. Several agents currently available in New Zealand provide adequate cover for commonly encountered pathogens. The choice of which agent to choose therefore rests more with their purchase and administration costs, as well as safety and efficacy data than simply susceptibility data alone.

Anti-Bacterial Agents↗

Aerobic bacterial isolates from choledochal bile at a tertiary hospital.

We studied the aerobic bacterial isolates from bile and their susceptibilty pattern in patients with biliary tract disease. Samples of bile collected during endoscopic retrograde cholangio pancreatography (ERCP) and surgery, were inoculated on standard media. Bacterial identification and susceptibility were done by standard techniques. A total of 209 samples were cultured, out of which 128 samples showed growth. Total number of isolates obtained was 221. Poly-microbial infection was detected in 67 patients. Predominant aerobic bacterial isolates obtained were Escherichia coli 30% (67), Klebsiella species 23.98% (53), Enterococcus species 12.21% (27). Multi-drug resistance was noted in 57%. Higher resistance rate was noted among Gram negative bacilli for ampicillin (92.4%), cephalexin (82.46%), ciprofloxacin (68.42%), piperacillin (64.33%). Sensitivity to meropenem was 90.64% and amikacin was 76.61%. Gram positive bacteria showed high resistance to gentamicin (39.53%). Sensitivity to ampicillin was 86.05% and penicillin was 81.4%. Vancomycin and teicoplanin showed 100% sensitivity. From our study we conclude that E. coli, Klebsiella species and Enterococcus species are common pathogens infecting biliary tract. Poly-microbial infection and multi-drug resistance warrants culture and sensitivity to guide antimicrobial therapy. We recommend combination of amikacin and ampicillin for empirical therapy at our institution.

Adolescent↗

[Screening of mixed crop disease-resistant bacterial inoculants from N2-fixing and P-solubilzing bacterial isolates].

In this paper, a total of 22 bacterial isolates capable of fixing nitrogen and/or solubilizing phosphorus were inoculated, and mixed incubated in 15 liquid media. The isolates were obtained from soil with selective media, and not purified. After grown for 48 hours, the mixed cultures were continuously inoculated to the same media, and then incubated under the same conditions. The procedures were repeated for 12 times, and 15 stable bacterial mixtures (SBM) were obtained. Each SBM was purified, which contained two or three bacterial species. Agar plate bacteriostasis test showed that 6 mixed cultures of the SBM exhibited significant inhibition effects to one or more plant pathogens of Verticillium dahliae, Rhizoctonia solani, and Pseudomonas syringae. It was found that C7 and C15 were the two SBM with most obvious antibiological effect. SMB C15 had the ability of nitrogen fixation, and C4, C5 C7, C9, C14 and C15 showed the ability of phosphorus solubilization. These results suggested a possible approach to develop stable, synergistic, and multi-functional antibiological agents.

Crops, Agricultural↗

Epidemiologic factors affecting antimicrobial resistance of common bacterial isolates.

The pattern of antimicrobial resistance of common bacterial isolates obtained from various groups of patients at a large tertiary-care center was compared with the pattern of resistance seen at a primary-care community hospital. At the tertiary-care center, significant differences in susceptibility were seen between pediatric and adult groups. In the tertiary-care center, the inpatients were more likely than the outpatients to have resistant staphylococcal and enterobacterial strains. Comparison of the overall resistance at the tertiary-care center and the primary-care hospital showed that resistance to cephalosporins, piperacillin, and aminoglycosides was significantly higher at the tertiary-care hospital than at the community hospital. Striking differences were noted in the resistance of nosocomial Enterobacter and Citrobacter isolates. Hospitals should be cautious in extrapolating nationwide data to their particular institutions.

Adolescent↗