Heterotrophic Plate Count Can Predict the Presence of Legionella spp. in Cooling Towers.

Legionella pneumophila (Lp) colonizes aquatic environments and is a potential pathogen to humans, causing outbreaks of Legionnaire's disease. It is mainly associated with contaminated cooling towers (CTs). Several regulations, including Spanish legislation (Sl), have introduced the analysis of heterotrophic plate count (HPC) bacteria and Legionella spp. (Lsp) in management plans to prevent and control Legionella outbreaks from CTs. The 2003 Sl for CTs (RD 865/2003) considered that concentrations of HPC bacteria ≤10,000 cfu/mL and of Lsp ≤100 cfu/L are safe; therefore, no action is required, whereas management actions should be implemented above these standards. We have investigated to what extent the proposed standard for HPC bacteria is useful to predict the presence of Lsp in cooling waters. For this, we analyzed Lsp and HPC concentrations, water temperature, and the levels of chlorine in 1376 water samples from 17 CTs. The results showed that in the 1138 water samples negative for Legionella spp. (LN), the HPC geometric mean was significantly lower (83 cfu/mL, p < 0.05) than in the positive Lsp. samples (135 cfu/mL). Of the 238 (17.3%) LP samples, 88.4% (210/238) were associated with values of HPC ≤10,000 cfu/mL and most of them showed HPC concentrations ≤100 (53.7%). In addition, a relatively low percentage of LP (28/238, 11.6%) samples were associated with HPC bacteria concentrations >10,000 cfu/mL, indicating that this standard does not predict the colonization risk for Legionella in the CTs studied. The present study has demonstrated that a threshold concentration ≤100 cfu/mL of HPC bacteria could better predict the higher concentration of Legionella in CTs, which will aid in preventing possible outbreaks.

Presence of Arcobacter spp. in environmental waters correlates with high levels of fecal pollution.

We investigated the presence of Arcobacter spp. in 205 water samples of freshwater, seawater and sewage in Spain. These bacteria were present in 55.1% of the samples (113/205) and were significantly associated for the first time with bacterial indicators of fecal pollution. The dominant species in the positive samples was Arcobacter butzleri (94%) followed by Arcobacter cryaerophilus (30%) and Arcobacter skirrowii (1.8%).

Fast detection of Salmonella Infantis with carbon nanotube field effect transistors.

In this paper we report a fast, sensitive and label-free biosensor for the selective determination of Salmonella Infantis. It is based on a field effect transistor (FET) in which a network of single-walled carbon nantotubes (SWCNTs) acts as the conductor channel. Anti-Salmonella antibodies were adsorbed onto the SWCNTs and subsequently the SWCNTs were protected with Tween 20 to prevent the non-specific binding of other bacteria or proteins. Our FET devices were exposed to increasing concentrations of S. Infantis and were able to detect at least 100 cfu/mL in 1h. To evaluate the selectivity of our FET devices, Streptococcus pyogenes and Shigella sonnei were tested as potential competing bacteria for Salmonella. At a concentration of 500 cfu/mL, neither Streptococcus nor Shigella interfered with the detection of Salmonella. Therefore, these devices could be used as useful label-free platforms to detect S. Infantis and, by using the suitable antibody, other bacteria or viruses.

Health risks of the occupational exposure to microbiological and chemical pollutants in a municipal waste organic fraction treatment plant.

Composting is a good alternative for the treatment of organic waste. However, an important amount of hazardous agents such as bioaerosols and volatile organic compounds may be released during the process. Therefore, the presence of microbiological and chemical pollutants emitted to air may mean a risk for the health of composting plants workers. We here report the results of an investigation aimed at evaluating the occupational exposure to chemical and biological agents for workers of an organic waste treatment facility (Montcada i Reixac, Catalonia, Spain). Total concentrations of bacteria and fungi (at 25 degrees C and 37 degrees C), including Aspergillus fumigatus, were determined on a 3-month basis in 4 areas of the composting plant (reception, sorting, composting and cogeneration halls). Non-cancer and cancer risks were assessed. Workers in the sorting cabins seemed to be the most exposed to pollutants. Consequently, the use of preventive measures, such as integrated P3 filter masks and gloves are highly recommended. On the other hand, the emission and dispersion of bioaerosols and particles should be minimized during the process through the application of additional measures, such as the humectation of waste and the installation of biofilters. The results of this study can be useful to elaborate occupational risk prevention programs for workers in composting plants.

Baseline levels of bioaerosols and volatile organic compounds around a municipal waste incinerator prior to the construction of a mechanical-biological treatment plant.

New waste management programs are currently aimed at developing alternative treatment technologies such as mechanical-biological treatment (MBT) and composting plants. However, there is still a high uncertainty concerning the chemical and microbiological risks for human health, not only for workers of these facilities, but also for the population living in the neighborhood. A new MBT plant is planned to be constructed adjacently to a municipal solid waste incinerator (MSWI) in Tarragona (Catalonia, Spain). In order to evaluate its potential impact and to differentiate the impacts of MSWI from those of the MBT when the latter is operative, a pre-operational survey was initiated by determining the concentrations of 20 volatile organic compounds (VOCs) and bioaerosols (total bacteria, gram-negative bacteria, fungi and Aspergillus fumigatus) in airborne samples around the MSWI. The results indicated that the current concentrations of bioaerosols (ranges: 382-3882, 18-790, 44-926, and <1-7 CFU/m(3) for fungi at 25 degrees C, fungi at 37 degrees C, total bacteria, and gram-negative bacteria, respectively) and VOCs (ranging from 0.9 to 121.2 microg/m(3)) are very low in comparison to reported levels in indoor and outdoor air in composting and MBT plants, as well in urban and industrial zones. With the exception of total bacteria, no correlations were observed between the environmental concentrations of biological agents and the direction/distance from the facility. However, total bacteria presented significantly higher levels downwind. Moreover, a non-significant increase of VOCs was detected in sites closer to the incinerator, which means that the MSWI could have a very minor impact on the surrounding environment.

Analysis of 16S rRNA gene mutations in a subset of Aeromonas strains and their impact in species delineation.

Characterization of 999 Aeromonas strains using a published 16S rDNA RFLP identification method showed that 8.1% of the strains produced unexpected (hereafter called "atypical") restriction patterns, making their identification uncertain. Atypical patterns were due to the presence of nucleotide polymorphisms among the rrn operons of the 16S rRNA gene (so-called microheterogeneities). Double sequencing signals at certain positions revealed the nucleotide composition was responsible for the microheterogeneities. Although the number of microheterogeneities was relatively low (0.06-0.66%), trees inferred from the 16S rRNA gene led either to a misidentification or to an inconclusive result for the majority of these strains. Strains with atypical patterns were, however, correctly identified using the rpoD gene sequences, as belonging to Aeromonas caviae, A. veronii, and A. media. All of them, but particularly the two former species, are associated with human disease. Microheterogeneities in 16S rRNA gene sequence were significantly (P 0.01) more prevalent in clinical than in environmental strains. This work also analyzed the effects of these microheterogeneities on the taxonomic position of the investigated strains. The results suggest the need for recording microheterogeneities in the 16S rRNA gene.

Effect of culture medium on the disk diffusion method for determining antifungal susceptibilities of dermatophytes.

We have evaluated a disk diffusion method to determine the activities of five drugs against 50 strains of dermatophytes and to assess the influence of the culture medium (antibiotic medium 3, high-resolution medium, and RPMI) on the inhibition zone diameters (IZD). There were no differences among the medium/drug combinations, except for itraconazole-RPMI, which showed the narrowest IZD.

Evaluation of disk diffusion method for determining eberconazole susceptibility of dermatophytes and influence of culture medium.

We have evaluated a disk diffusion method to determine the activity of eberconazole against 50 strains of dermatophytes by testing three culture media (RPMI, antibiotic medium 3, and high resolution). No differences were found among the results obtained with the three media. A significant correlation between disk diffusion and microdilution methods was observed with AM3.

In vitro antifungal susceptibilities of Sporothrix schenckii in two growth phases.

We have determined the antifungal susceptibilities of 34 clinical isolates of the dimorphic fungus Sporothrix schenckii to 11 drugs using a microdilution method. In general, the type of growth phase (mycelial or yeast) and the temperature of incubation (30 or 35 degrees C) exerted a significant influence on the MICs.

In vitro activities of the new antifungal drug eberconazole and three other topical agents against 200 strains of dermatophytes.

We have compared the in vitro activity of the new antifungal drug eberconazole with those of three other topical antifungal agents, clotrimazole, ketoconazole, and miconazole, against 200 strains of dermatophytes. MICs were determined by a microdilution method with optimal conditions determined in a previous study (an inoculum of 10(4) CFU/ml, an incubation temperature of 28 degrees C, an incubation period of 7 days, and a MIC endpoint of 100% inhibition of growth). In general, the four drugs tested showed low MICs. However, eberconazole was more active (P < 0.05) than the other three drugs against the majority of the species tested. Eberconazole represents an advantageous alternative for dermatophytoses where a topical therapy is required.

Collaborative evaluation of optimal antifungal susceptibility testing conditions for dermatophytes.

A multicenter study was conducted to define the most suitable testing conditions for antifungal susceptibility of dermatophytes. Broth microdilution MICs of clotrimazole, itraconazole, and terbinafine were determined in three centers against 60 strains of dermatophytes. The effects of inoculum density (ca. 10(3) and 10(4) CFU/ml), incubation time (3, 7, and 14 days), endpoint criteria for MIC determination (complete [MIC-0] and prominent [MIC-2] growth inhibition), and incubation temperature (28 and 37 degrees C) on intra- and interlaboratory agreement were analyzed. The optimal testing conditions identified were an inoculum of 10(4) CFU/ml, a temperature of incubation of 28 degrees C, an incubation period of 7 days, and MIC-0.