Predicting Legionella contamination in cooling towers and evaporative condensers from microbiological and physicochemical parameters.

BACKGROUND: Inhalation of Legionella-containing aerosols generated by cooling towers (CT) and evaporative condensers (EC) where water risk management is not performed correctly has been linked to a high percentage of community outbreaks of Legionnaires' disease (LD). Likewise, microbiological and physicochemical characteristics of the water in these facilities have been associated with this bacterium. The main aim of this study was to assess the risk of Legionella colonization in CT and EC based on the data for microbiological and physicochemical water quality provided by the Environmental Health Department and Laboratory of the City Council of L'Hospitalet de Llobregat (Barcelona, Spain). METHODS: Legionella was analysed in 789 samples collected from 127 CT and EC in 46 companies in Catalonia from 2002 to 2019. A two-step logistic regression analysis was carried out to assess the risk of colonization by Legionella in the studied facilities according to the microbiological (aerobic heterotrophic bacteria) and physicochemical (pH, alkalinity, hardness, turbidity, conductivity, total iron and Langelier Index) water parameters. The optimal cut-off points for the water parameters predictive of Legionella contamination were defined as the values on the receiver operating characteristic (ROC) curve where sensitivity and specificity were jointly maximized. RESULTS: Legionella was isolated in 8.49% of the 789 analysed samples, 22.39% of which were heavily contaminated (with counts higher than 1.0 × 104 CFU/l). L. pneumophila was isolated in 82.09% of the samples, with 41.82% belonging to serogroup 1. Logistic regression analysis revealed that aerobic heterotrophic bacteria concentrations ≥6.90 × 102 CFU/ml [Odds ratios (OR) (95% CI) = 3.56 (1.39-9.43), p = 0.01], a pH ≥ 8.70 [OR (95% CI) = 3.60 (1.34-10.09), p = 0.01], and water hardness ≥5.72 × 102 mg/l [OR (95% CI) = 6.30 (2.34-18.56), p < 0.001] were each independently associated with a higher risk of CT and EC colonization by Legionella. CONCLUSIONS: The present study shows the importance of risk assessment for improving the control measures aimed at preventing or reducing Legionella populations in CT and EC, thus minimizing potential dangers for public health.

Exposure of zebrafish to elevated temperature induces sex ratio shifts and alterations in the testicular epigenome of unexposed offspring.

Accumulating evidence shows that environmental changes can affect population sex ratios through epigenetic regulation of gene expression in species where sex depends on both genetic and environmental cues. Sometimes, altered sex ratios persist in the next generation even when the environmental cue is no longer present (a multigenerational effect). However, evidence of transgenerational effects (i.e., beyond the first non-exposed generation), which tend to be paternally transmitted, is scarce and a matter of debate. Here, we used the AB strain of zebrafish, where sex depends on both genetic and environmental influences, to study possible multi- (to the F1) and transgenerational (to the F2) effects of elevated temperature during the critical period of sex differentiation. From eight initial different families, five were selected in order to capture sufficient variation between the sex ratio of the control group (28 °C) and the group exposed to elevated (35 °C) temperature only at the parental (P) generation. Results showed a consistent increase in the proportion of males in the P generation in all five families as a result of heat treatment. Sex ratios were then determined in the F1 and F2 offspring derived from both above groups, which were all raised at 28 °C. A persisting male-skewed sex ratio in the 35°C-derived, unexposed offspring of the F1 generation was observed in three families, denoting family-dependent multigenerational effects. However, no transgenerational effects were observed in the F2 generation of any family. DNA methylation was also assessed in the testis of P, F1 and F2 males derived from exposed and non-exposed fathers and grandfathers. DNA methylation was significantly decreased only in the testis of the 35°C-derived males in the F1 generation but not of the F2 generation and, surprisingly, neither in the 35°C-exposed males of the P generation. Taken together, our results show great interfamily variation, not only in sex ratio response to elevated temperature, but also on its multigenerational effects, denoting a strong influence of genetics. Alterations in the testicular epigenome in F1 males calls for attention to possible, previously unnoticed, effects of temperature in the unexposed offspring of heat-exposed parents in a global warming scenario.

Traceability of different brands of bottled mineral water during shelf life, using PCR-DGGE and next generation sequencing techniques.

Natural mineral waters contain indigenous bacteria characteristic of each spring source. Once bottled, these communities change over time until the water is consumed. Bottle material is believed to play a major role in the succession of these populations, but very few studies to date have evaluated the effect of this material on bacterial communities. In this study, we examined the microbial community structure of three natural mineral waters over 3 months after bottling in glass and polyethylene terephthalate (PET) bottles. To this end, we used culture-dependent (heterotrophic plate count) and culture-independent methods (16S rRNA massive gene sequencing, denaturing gradient gel electrophoresis (DGGE) and fluorescent microscopy with vital dyes). Total and viable cell counts increased by around 1-2 log10 units between 1 and 2 weeks after bottling and then remained constant over 3 months for all waters regardless of the bottle material. DGGE fingerprints and 16S rRNA massive sequencing analysis both indicated that different communities were established in the waters two weeks after bottling in the different bottle materials. In conclusion, no differences in total, viable and culturable bacteria counts were observed between mineral waters bottled with PET or glass during shelf life storage. Nevertheless, in spite of changes in the communities, each water brand and material presented a distinct microbial community structure clearly distinguishable from the others, which could be interesting for traceability purposes.

Long-term risk of cardiovascular events after detecting silent coronary atheromatosis using computed tomography.

BACKGROUND: Coronary calcium score (CCS) and coronary computed tomography angiography (CTA) assessments using multidetector computed tomography are invaluable for atheromatosis screening. We studied their usefulness in cardiovascular risk assessments, and compared evaluations using the Systematic COronary Risk Evaluation (SCORE) algorithm with those from CTA and CSS assessments in terms of their ability to predict cardiovascular events in Mediterranean patients. PATIENTS AND METHODS: Two hundred and sixty-six asymptomatic patients whose mean age was 55.4 years, 89.5% of whom were men, were evaluated using CTA and CCS and followed for more than 10 years. The CTA and CCS risk predictions were compared with those determined using the SCORE algorithm designed for low-risk populations. RESULTS: Coronary lesions were present in 140 (53.4%) patients. Of the lesions, 17% were noncalcified, 17% were mixed, and 66% were calcified; in addition, 24.2% of the patients who had lesions had cardiovascular events during follow-up (P<0.00001), but just 2.9% of the patients without lesions. Detection of atheromatosis using computed tomography was associated with an increased risk of cardiovascular disease events at more than 10 years [odds ratio (OR): 6.828; 95% confidence interval (CI): 2.001-23.305; P=0.002]. This OR was higher than that obtained for intermediate-risk individuals (OR: 4.818; 95% CI: 1.360-17.075; P=0.015) and lower than that determined for high-risk individuals (OR: 9.395; 95% CI: 2.489-35.460; P=0.001) using the SCORE algorithm, and higher that that determined for CCS assessments (OR: 3.916; 95% CI: 1.572-9.751; P=0.03). More cardiovascular events were associated with higher amounts of calcium. CONCLUSION: The detection of atheromatosis using the CCS and CTA was associated with an increased risk of cardiovascular events at more than 10 years. CTA and CCS assessments had a higher OR than that associated with assessments of patients at intermediate risk using the SCORE algorithm.

Identifying Prognostic Factors for Well-Differentiated Metastatic Pancreatic Neuroendocrine Tumours: A Retrospective International Multicentre Cohort Study.

Pancreatic neuroendocrine tumours (pNETs) represent rare neoplasms of all NETs often presenting without functional activity. Many sporadic non-functioning pNET patients are already metastatic at the time of diagnosis, and the therapeutic approach to such patients is mostly palliative. In this international, multicentre, retrospective cohort study, we assessed the prognostic value of a set of anthropometric, clinical, biochemical, radiological and pathological parameters at baseline and the impact of the therapeutic strategies on the survival of patients with sporadic grade 1/2, stage IV, non-functioning pNETs. Three hundred and twelve consecutive patients diagnosed between 1993 and 2010 were included. The median overall survival (OS) was 6.6 years and survival at 5 and 10 years was 62 and 34% respectively. On univariate analysis, Eastern Cooperative Oncology Group (ECOG) status ≥2, grade 2, bilobar hepatic metastases, synchronous metastases, and high chromogranin A, alkaline-phosphatase and lactic-dehydrogenase were associated with a significant reduction of OS. Palliative/curative surgery and loco-regional hepatic interventions were significant factors improving OS. On multivariate analysis, ECOG status ≥2, synchronous metastases, Ki-67 ≥10%, and high alkaline-phosphatase correlated significantly with an increased risk of death. Both palliative/curative surgery and loco-regional hepatic interventions had a positive impact on OS. Although most parameters did not prove to be independent OS predictors at multivariate analysis, they showed a tendency towards that. Future prospective studies including larger patient populations may give greater clarity. We believe the integration of these parameters has the potential to provide a reliable prognostic score for the stratification of patients with sporadic well-differentiated metastatic non-functioning pNETs.

Canalization technique to obtain deep tissue biopsy of gastrointestinal subepithelial tumors as an alternative to conventional known techniques.

BACKGROUND AND OBJECTIVES: The most accurate technology to detect and diagnose subepithelial tumors (SETs) is the endoscopic ultrasonography (EUS) combined with puncture techniques, such as the endoscopic ultrasonography-guided fine-needle aspiration (EUS-FNA) or the endoscopic ultrasonography-guided fine-needle biopsy. Going further in the improvement of the results of tumor samples obtained endoscopically to diagnose the SETs, the canalization technique guided by miniprobes (MPs) to obtain biopsies of SET could be an alternative to EUS-FNA. The objective of this study is to analyze the results of samples obtained by this procedure. MATERIALS AND METHODS: A multicenter, retrospective study of a review of a database of 32 consecutive patients with a SET in the digestive tract, from 2000 to 2015 was conducted. All patients underwent EUS-performed by MP, to define the size, internal echostructure, and layer of origin of tumor. Once the echostructure was defined, it proceeded to the canalization technique to arrive to the tumor tissue. RESULTS: The average diameter of SETs in this series (32 patients) was about 21.6±11 mm (range: 5-41 mm). The diagnostic accuracy was 28/32, 87.50% (Confidence interval 95%: 76.04%-98.99%), and there were no major complications. All procedures were performed on outpatients, none of which required additional hospitalization. The 50% of patients were operated or endoscopically resected and in all cases, the previous pathological diagnosis was confirmed. CONCLUSIONS: This is a feasible, safe, and effective procedure that allows to access to inside of SET to obtain deep biopsies. Tumor samples obtained by deep biopsy, with prior performing of the canalization technique guided by MP, were sufficient for histopathological and immunohistochemical diagnosis and similar to those obtained with other known methods (FNA Trucut, ProCore®, etc.). However, more prospective comparative studies with a larger number of patients and different specialists carrying out the procedure to reach a higher statistical significance are necessary.

Use of non-linear mixed-effects modelling and regression analysis to predict the number of somatic coliphages by plaque enumeration after 3 hours of incubation.

The present study aimed to establish the kinetics of the appearance of coliphage plaques using the double agar layer titration technique to evaluate the feasibility of using traditional coliphage plaque forming unit (PFU) enumeration as a rapid quantification method. Repeated measurements of the appearance of plaques of coliphages titrated according to ISO 10705-2 at different times were analysed using non-linear mixed-effects regression to determine the most suitable model of their appearance kinetics. Although this model is adequate, to simplify its applicability two linear models were developed to predict the numbers of coliphages reliably, using the PFU counts as determined by the ISO after only 3 hours of incubation. One linear model, when the number of plaques detected was between 4 and 26 PFU after 3 hours, had a linear fit of: (1.48 × Counts3 h + 1.97); and the other, values >26 PFU, had a fit of (1.18 × Counts3 h + 2.95). If the number of plaques detected was <4 PFU after 3 hours, we recommend incubation for (18 ± 3) hours. The study indicates that the traditional coliphage plating technique has a reasonable potential to provide results in a single working day without the need to invest in additional laboratory equipment.