Cadmium phytoextraction by Helianthus annuus (sunflower), Brassica napus cv Wichita (rapeseed), and Chyrsopogon zizanioides (vetiver).

The use of phytoextraction plant species to accumulate soil metals into harvestable plant parts is a method used for managing soils with high cadmium (Cd). We evaluated three Cd accumulating species recently recommended for such use in cacao farms where Cd removal is needed to maintain markets: Helianthus annuus (sunflower), Brassica napus (rapeseed), and Chyrsopogon zizanioides (vetiver). Plants were grown in two greenhouse pot experiments with different Cd-spiked growth media: (sand plus perlite) and a natural soil. Plant total Cd and Cd uptake in shoot biomass of all species, across both experiments, increased linearly with increasing amounts of added Cd. Rapeseed had the highest plant total Cd and sunflower had the highest Cd uptake in shoot biomass. The highest application of Cd corresponded to the highest plant total Cd and shoot biomass Cd uptake, regardless of species. The bioconcentration factor (BCF) for each species increased in a curvilinear manner with added Cd, with maximum BCF values for plants grown in the sand and perlite matrix at 2.5 mg kg-1 added Cd and in the natural soil at 5.0 mg kg-1 added Cd. We conclude that the Cd uptake (shoot biomass only) capability of the three species examined is greatest for sunflower given its increased uptake with Cd additions, its BCF value > 1, and lack of observed visual Cd toxicity symptoms, fungus and insect damage. Although these species had BCF >1, the potential annual removal of Cd would have been too small to support a meaningful phytoextraction practice.

Optimal inoculation methods and quality control for the NCCLS oxacillin agar screen test for detection of oxacillin resistance in Staphylococcus aureus.

To define more precisely the inoculation methods to be used in the oxacillin screen test for Staphylococcus aureus, we tested agar screen plates prepared in house with 6 microg of oxacillin/ml and 4% NaCl using the four different inoculation methods that would most likely be used by clinical laboratories. The organisms selected for testing were 19 heteroresistant mecA-producing strains and 41 non-mecA-producing strains for which oxacillin MICs were near the susceptible breakpoint. The inoculation method that was preferred by all four readers and that resulted in the best combination of sensitivity and specificity was a 1-microl loopful of a 0.5 McFarland suspension. A second objective of the study was to then use this method to inoculate plates from five different manufacturers of commercially prepared media. Although all commercial media performed with acceptable sensitivity compared to the reference lot, one of the commercial lots demonstrated a lack of specificity. Those lots of oxacillin screen medium that fail to grow heteroresistant strains can be detected by using S. aureus ATCC 43300 as a positive control in the test and by using transmitted light to carefully examine the plates for any growth. However, lack of specificity with commercial lots may be difficult to detect using any of the current quality control organisms.

Detection of Klebsiella pneumoniae and Escherichia coli strains producing extended-spectrum beta-lactamases.

Plasmids encoding extended-spectrum beta-lactamases of the TEM, SHV, and AmpC families were introduced into common Escherichia coli and Klebsiella pneumoniae hosts to create a homogeneous panel for evaluating the abilities of five test systems to detect resistance to eight beta-lactam antibiotics. Although MICs, as determined by agar dilution or E test strips, were increased and disk diffusion zone diameters were diminished, breakpoints for resistance were often not reached, and neither approach was sensitive in detecting resistance to oxyimino-beta-lactams. The MicroScan 18-h microdilution or Vitek rapid automated procedures were similarly insensitive. Ceftazidime was the best single test antibiotic for detecting extended-spectrum beta-lactamase production. beta-Lactamases TEM-7 and TEM-12 were particularly difficult to detect. Because of such difficulties, the prevalence of extended-spectrum beta-lactamases is likely to be greater than is currently appreciated.

Detection of penicillin and extended-spectrum cephalosporin resistance among Streptococcus pneumoniae clinical isolates by use of the E test.

Increasing penicillin resistance and the initial recognition of resistance to extended-spectrum cephalosporins among Streptococcus pneumoniae isolates have placed greater emphasis on accurate methods for susceptibility testing of clinical isolates. This study has evaluated the use of the E test (AB Biodisk NA, Piscataway, N.J.) for the detection of penicillin and cefotaxime resistance among 147 pneumococcal clinical isolates in three geographically separate laboratories. These included 42 penicillin-resistant (MIC, > or = 2 micrograms/ml) and 14 cefotaxime-resistant (defined here as an MIC of > or = 2 micrograms/ml) isolates. E test strips were applied to the surface of Mueller-Hinton sheep blood agar plates and incubated at 35 degrees C in 5% CO2 for 20 to 24 h. E test MICs were compared with MICs determined with lysed horse blood-supplemented Mueller-Hinton broth in a microdilution format as recommended by the National Committee for Clinical Laboratory Standards. Penicillin MICs agreed within one log2 dilution for 136 of 147 (92.5%) isolates, and cefotaxime MICs agreed within one log2 dilution for 142 of 147 (96.6%) isolates. No very major or major interpretive errors occurred with either penicillin or cefotaxime E test MIC results. There were 9.5 and 5.4% minor interpretive category errors with penicillin and cefotaxime E test MICs, respectively. These data indicate that the E test represents a convenient and reliable method for the detection of penicillin or cephalosporin resistance in pneumococci.

Detection of enterococcal high-level aminoglycoside resistance with MicroScan freeze-dried panels containing newly modified medium and Vitek Gram-Positive Susceptibility cards.

Both conventional and modified MicroScan Type 5 panels and Vitek Gram-Positive Susceptibility cards were compared with agar dilution screen plates for their abilities to detect high-level resistance to gentamicin and streptomycin in 235 enterococcal isolates, including 167 Enterococcus faecalis and 63 E. faecium isolates. The modified Type 5 panels contained dextrose-phosphate broth instead of Mueller-Hinton broth in their high-level-resistance screen wells. The sensitivities for detection of gentamicin and streptomycin high-level resistance were 100 and 100% (E. faecalis) and 100 and 94% (E. faecium) for the modified MicroScan panels, 100 and 89% (E. faecalis) and 100 and 98% (E. faecium) for the conventional MicroScan panels, and 81 and 86% (E. faecalis) and 85 and 94% (E. faecium) for the Vitek cards. All specificities were 100% except for the Vitek cards with streptomycin, where it was 96%. Isolates that showed resistance on the streptomycin agar screen plates were rescreened on plates containing 32,000 micrograms/ml to detect ribosomally mediated resistance. For all three systems, every failure to detect streptomycin high-level resistance occurred in isolates with enzymatic, not ribosomal, resistance. The modified MicroScan Type 5 panels are a suitable method for detecting enterococcal high-level resistance to gentamicin and streptomycin. The Vitek cards are too insensitive for this purpose.

Species identities of enterococci isolated from clinical specimens.

Conventional tests and commercially available systems were used to determine the species identities of clinical isolates of enterococci. Strict adherence to the conventional test scheme of Facklam and Collins (R. R. Facklam and M. D. Collins, J. Clin. Microbiol. 27:731-734, 1989) resulted in the misidentification of lactose-negative Enterococcus faecalis isolates as Enterococcus solitarius, but this problem was overcome by the application of additional tests. The commercially available systems tested were unable to recognize some of the more recently described enterococcal species. E. faecalis accounted for 87.1% of 302 consecutive isolates. Enterococcus faecium (8.6%), Enterococcus avium (0.7%), Enterococcus durans (0.3%), Enterococcus gallinarum (1.0%), Enterococcus casseliflavus (1.0%), Enterococcus hirae (0.3%), and Enterococcus raffinosus (0.3%) isolates were also identified. None of the isolates produced beta-lactamase, but 15.4% of 235 isolates tested, including 1 strain of E. gallinarum, displayed high-level resistance to gentamicin.