A curated collection of Klebsiella metabolic models reveals variable substrate usage and gene essentiality.

The Klebsiella pneumoniae species complex (KpSC) is a set of seven Klebsiella taxa that are found in a variety of niches and are an important cause of opportunistic health care-associated infections in humans. Because of increasing rates of multi-drug resistance within the KpSC, there is a growing interest in better understanding the biology and metabolism of these organisms to inform novel control strategies. We collated 37 sequenced KpSC isolates isolated from a variety of niches, representing all seven taxa. We generated strain-specific genome-scale metabolic models (GEMs) for all 37 isolates and simulated growth phenotypes on 511 distinct carbon, nitrogen, sulfur, and phosphorus substrates. Models were curated and their accuracy was assessed using matched phenotypic growth data for 94 substrates (median accuracy of 96%). We explored species-specific growth capabilities and examined the impact of all possible single gene deletions using growth simulations in 145 core carbon substrates. These analyses revealed multiple strain-specific differences, within and between species, and highlight the importance of selecting a diverse range of strains when exploring KpSC metabolism. This diverse set of highly accurate GEMs could be used to inform novel drug design, enhance genomic analyses, and identify novel virulence and resistance determinants. We envisage that these 37 curated strain-specific GEMs, covering all seven taxa of the KpSC, provide a valuable resource to the Klebsiella research community.

Insights on trends in the management of metrological analysis of devices in blood and transfusion medicine services: Verification based on their fitness for purpose and suitability for their respective applications.

In the field of blood and transfusion medicine, European Directorate for the Quality of Medicines & HealthCare recommends practices for metrological devices. Monitoring and measurement resources, as well as their permitted metrological limits, must be considered in this regard. We must understand variables such as metrological domains, the International System of Units, metrological traceability, and metrological requirements and controls in the field of transfusions in order to understand these metrological devices. An approach to metrological analysis and subsequent actions was constructed using flowcharts. As part of this methodology, a metrological device was selected, along with regulatory and normative requirements. A calibration and a test were conducted to demonstrate the application of the figure based on current recommendations. Current recommendations are consistent with the suggested ROC approach. The metrological devices must be verified based on their suitability for their respective applications (fitness-for-purpose).

Resurgence of Bordetella pertussis, including one macrolide-resistant isolate, France, 2024.

As other European countries, France is experiencing a resurgence of pertussis in 2024. Between 1 January and 31 May 2024, 5,616 (24.9%) positive Bordetella pertussis qPCR tests were identified, following a 3-year period of almost null incidence. Of 67 cultured and whole genome sequenced B. pertussis isolates, 66 produced pertactin and 56 produced FIM2, in contrast to pre-COVID-19 years. One isolate of genotype Bp-AgST4 was resistant to macrolides. Pertussis resurgence may favour isolates that produce FIM2 and pertactin.

Comparative phylogenomics of ESBL-, AmpC- and carbapenemase-producing Klebsiella pneumoniae originating from companion animals and humans.

BACKGROUND: WHO considers ESBL- and carbapenemase-producing Klebsiella pneumoniae a major global concern. In animals, ESBL- and carbapenemase-producing K. pneumoniae of human-related ST11, ST15 and ST307 have been reported, but not in the context of large WGS-based One Health investigations. OBJECTIVES: To perform comparative phylogenomics on a large collection of multidrug-resistant (MDR) K. pneumoniae recovered from diseased companion animals and humans. METHODS: MDR K. pneumoniae (n = 105) recovered from companion animals in France during 2010-18 were phenotypically characterized. All isolates were whole-genome sequenced using the NovaSeq technology and phylogenomic analysis across animal and human K. pneumoniae was performed using appropriate pipelines. RESULTS: bla CTX-M-15, blaDHA-1 and blaOXA-48 were strongly associated with IncFIIk, IncR and IncL plasmids, respectively. When compared with human K. pneumoniae genomes, four groups of closely related French human and animal isolates belonging to ST11, ST15 and ST307 were detected, suggesting the circulation of clones between the human and animal sectors at country level. A large cluster of 31 ST11-KL105 animal isolates from France and Switzerland suggested it corresponds to a sub-lineage that is particularly well-adapted to the animal host. CONCLUSIONS: This study demonstrates the spread of blaCTX-M-15-carrying ST15 and ST307, and blaDHA-1-carrying ST11 K. pneumoniae clones in animal populations. ST11 was the main vector of blaOXA-48/IncL, despite the absence of carbapenem use in French animals. Comparative phylogenomics suggests cross-transmission of K. pneumoniae sub-lineages more prone than others to colonize/infect the animal host. Our data also evidenced the emergence of convergent hypervirulent and MDR K. pneumoniae in animals.

The success of particular Acinetobacter baumannii clones: accumulating resistance and virulence inside a sugary shield.

BACKGROUND: In Portugal, carbapenem-resistant Acinetobacter baumannii (CRAB) has been associated with ST98, ST103 and ST208 (Oxford Scheme, Oxf) and a clone has usually been associated with a particular period of time. These clonal shifts were primarily explained by an increased antimicrobial resistance profile. Here we explore genomic and biochemical differences among these and more recent clones, which could further explain the diversity and evolution of this species. METHODS: A total of 116 CRAB isolates (2010-15), together with representatives of a previously described CRAB collection (4 isolates, 2001-06) were characterized by attenuated total reflection Fourier transform infrared spectroscopy (FTIR-ATR) and MLST. Representatives of different FTIR-ATR/MLST clusters were selected for WGS (n = 13), which allowed the in silico extraction of resistance and virulence genes, capsule locus and SNP analysis. RESULTS: A. baumannii clonal shifts of OXA-58-producing ST103Oxf (2001-04), OXA-40-producing ST98Oxf (2002-06), OXA-23-producing ST208Oxf (2006-10) and OXA-23-producing ST218Oxf (2010-15) were accompanied by an increase in AMR genes and virulence factors. FTIR-ATR clustering was congruent with sugar composition predicted from the capsular locus: a fucosamine cluster comprising ST98Oxf, ST103Oxf and a single ST218Oxf isolate; a pseudaminic acid cluster of ST208Oxf and ST1557Oxf isolates; and legionaminic acid, resembling the sialic acid from mammalian cells, in a cluster comprising ST218Oxf isolates. The whole-genome phylogenetic tree was congruent with MLST, with isolates presenting 5-28 938 SNPs. ST208Oxf and ST218Oxf presented ∼1900 SNPs while ST103Oxf and ST1557Oxf showed a greater number of SNPs (∼28 000). CONCLUSIONS: Clonal shifts of CRAB were promoted, in our country, by consecutive virulence and AMR gene pool enlargement, together with features increasing pathogen-host adaptation. Worldwide dominance of ST218Oxf is supported by the combination of high AMR and virulence levels.

Aerobic growth physiology of Saccharomyces cerevisiae on sucrose is strain-dependent.

Present knowledge on the quantitative aerobic physiology of the yeast Saccharomyces cerevisiae during growth on sucrose as sole carbon and energy source is limited to either adapted cells or to the model laboratory strain CEN.PK113-7D. To broaden our understanding of this matter and open novel opportunities for sucrose-based biotechnological processes, we characterized three strains, with distinct backgrounds, during aerobic batch bioreactor cultivations. Our results reveal that sucrose metabolism in S. cerevisiae is a strain-specific trait. Each strain displayed distinct extracellular hexose concentrations and invertase activity profiles. Especially, the inferior maximum specific growth rate (0.21 h-1) of the CEN.PK113-7D strain, with respect to that of strains UFMG-CM-Y259 (0.37 h-1) and JP1 (0.32 h-1), could be associated to its low invertase activity (0.04-0.09 U/mgDM). Moreover, comparative experiments with glucose or fructose alone, or in combination, suggest mixed mechanisms of sucrose utilization by the industrial strain JP1, and points out the remarkable ability of the wild isolate UFMG-CM-259 to grow faster on sucrose than on glucose in a well-controlled cultivation system. This work hints to a series of metabolic traits that can be exploited to increase sucrose catabolic rates and bioprocess efficiency.

Female sexual function and quality of life after pelvic floor surgery: a prospective observational study.

INTRODUCTION AND HYPOTHESIS: Knowledge about the impact of pelvic floor surgery on sexual function is limited and inconsistent. A prospective study assessed the impact of surgery for prolapse (POP) or stress urinary incontinence (SUI) on sexual function and determined the biopsychosocial predictors for changes in sexual function after surgery. MATERIALS AND METHODS: Sexually active women scheduled for correction of POP and/or SUI were recruited over a 6-month period. Consenting participants were asked to complete the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ-12) to evaluate sexual function. The King's Health Questionnaire and Prolapse Quality-of-Life Questionnaire were used to assess UI and POP symptoms and their impact on quality of life (QoL), respectively. Women were reviewed over a period of 1 year post-surgery. RESULTS: Ninety-one patients were followed up over 1 year. After stratification into only or both SUI and POP surgery, global sexual function improved significantly in each group (p < 0.001). The improvement in the overall QoL score after pelvic surgery showed a significant correlation with the improvement in global sexual function (SUI group: r = - 0.38, p < 0.01; POP group: r = - 0.44, p < 0.05). For women undergoing SUI surgery, only educational level and prior hysterectomy had a significant association with improved sexual function. For women undergoing POP surgery with or without SUI surgical repair, no sociodemographic characteristics were preditive of improvement of sexual function. CONCLUSIONS: Significantly improved sexual function was observed 1 year after pelvic floor surgery, and the improvement was predicted by other social and physical factors in addition to normal functional anatomy.

Prevalence of Campylobacter spp. in Chicken Carcasses in Slaughterhouses from South of Brazil.

Campylobacteriosis has become common cause of diarrhea in humans and is associated with Guillain-Barré Syndrome, Reactive Arthritis and Irritable Bowel Syndrome is caused mainly by contaminated food and water intake in which the majority occurs from manipulation, preparation and consumption of poultry meat. The aim of this study was to estimate the prevalence of Campylobacter in chicken carcasses from slaughterhouses located in the states of Parana, Santa Catarina and Rio Grande do Sul in the South of Brazil. The samples were analyzed for Campylobacter enumeration using the ISO method 10272-2 and the species C. jejuni and C. coli, important for public health, were identified through Maldi-TOF mass spectrometry. From July 2017 to July 2018, 816 samples were analyzed, indicating the prevalence of 35.84%, with higher occurrence of C. jejuni (78.47%). No difference in prevalence was observed in relation to the size of the slaughterhouses. However, significant differences were noted among the three states in the southern region of the country, with the lowest prevalence being observed in Parana. The results reinforce the need to advance in the implementation of strategies to control this pathogen in the country, in order to safeguard consumer's health and contribute for the maintenance of Brazil's position in the international poultry meat market.

The ZKIR Assay, a Real-Time PCR Method for the Detection of Klebsiella pneumoniae and Closely Related Species in Environmental Samples.

Klebsiella pneumoniae is of growing public health concern due to the emergence of strains that are multidrug resistant, virulent, or both. Taxonomically, the K. pneumoniae complex ("Kp") includes seven phylogroups, with Kp1 (K. pneumoniaesensu stricto) being medically prominent. Kp can be present in environmental sources such as soils and vegetation, which could act as reservoirs of animal and human infections. However, the current lack of screening methods to detect Kp in complex matrices limits research on Kp ecology. Here, we analyzed 1,001 genome sequences and found that existing molecular detection targets lack specificity for Kp. A novel real-time PCR method, the ZKIR (zur-khe intergenic region) assay, was developed and used to detect Kp in 96 environmental samples. The results were compared to a culture-based method using Simmons citrate agar with 1% inositol medium coupled to matrix-assisted laser desorption ionization-time of flight mass spectrometry identification. Whole-genome sequencing of environmental Kp was performed. The ZKIR assay was positive for the 48 tested Kp reference strains, whereas 88 non-Kp strains were negative. The limit of detection of Kp in spiked soil microcosms was 1.5 × 10-1 CFU g-1 after enrichment for 24 h in lysogeny broth supplemented with ampicillin, and it was 1.5 × 103 to 1.5 × 104 CFU g-1 directly after soil DNA extraction. The ZKIR assay was more sensitive than the culture method. Kp was detected in 43% of environmental samples. Genomic analysis of the isolates revealed a predominance of phylogroups Kp1 (65%) and Kp3 (32%), a high genetic diversity (23 multilocus sequence types), a quasi-absence of antibiotic resistance or virulence genes, and a high frequency (50%) of O-antigen type 3. This study shows that the ZKIR assay is an accurate, specific, and sensitive novel method to detect the presence of Kp in complex matrices and indicates that Kp isolates from environmental samples differ from clinical isolates.IMPORTANCE The Klebsiella pneumoniae species complex Kp includes human and animal pathogens, some of which are emerging as hypervirulent and/or antibiotic-resistant strains. These pathogens are diverse and classified into seven phylogroups, which may differ in their reservoirs and epidemiology. Proper management of this public health hazard requires a better understanding of Kp ecology and routes of transmission to humans. So far, detection of these microorganisms in complex matrices such as food or the environment has been difficult due to a lack of accurate and sensitive methods. Here, we describe a novel method based on real-time PCR which enables detection of all Kp phylogroups with high sensitivity and specificity. We used this method to detect Kp isolates from environmental samples, and we show based on genomic sequencing that they differ in antimicrobial resistance and virulence gene content from human clinical Kp isolates. The ZKIR PCR assay will enable rapid screening of multiple samples for Kp presence and will thereby facilitate tracking the dispersal patterns of these pathogenic strains across environmental, food, animal and human sources.

Fourier transform infrared (FT-IR) spectroscopy typing: a real-time analysis of an outbreak by carbapenem-resistant Klebsiella pneumoniae.

The spread of multidrug-resistant (MDR) Klebsiella pneumoniae in the nosocomial setting represents a big challenge to infection control teams. We have recently developed a simple spectroscopic-based method with excellent accuracy, turnaround time and cost-effectiveness (Rodrigues et al. mSystems 2020) for bacterial typing. Here, we applied our method in a real clinical context to support early identification of an outbreak involving KPC-3-producing K. pneumoniae ST147 isolates. Our results further support that attenuated total reflectance Fourier transform infrared (FT-IR) spectroscopy can provide enough information to support early and adequate infection control measures and therapeutic choices in the context of nosocomial outbreaks and hospital surveillance.

A comparative in silico linear B-cell epitope prediction and characterization for South American and African Trypanosoma vivax strains.

Trypanosoma vivax is a parasite widespread across Africa and South America. Immunological methods using recombinant antigens have been developed aiming at specific and sensitive detection of infections caused by T. vivax. Here, we sequenced for the first time the transcriptome of a virulent T. vivax strain (Lins), isolated from an outbreak of severe disease in South America (Brazil) and performed a computational integrated analysis of genome, transcriptome and in silico predictions to identify and characterize putative linear B-cell epitopes from African and South American T. vivax. A total of 2278, 3936 and 4062 linear B-cell epitopes were respectively characterized for the transcriptomes of T. vivax LIEM-176 (Venezuela), T. vivax IL1392 (Nigeria) and T. vivax Lins (Brazil) and 4684 for the genome of T. vivax Y486 (Nigeria). The results presented are a valuable theoretical source that may pave the way for highly sensitive and specific diagnostic tools.

Fourier transform infrared spectroscopy: unlocking fundamentals and prospects for bacterial strain typing.

The need to identify highly related bacterial strains is ancient in clinical, industrial, or environmental microbiology. Strategies based on different phenotypic and genotypic principles have been used since the early 1930s with variable outcomes and performances, accompanying the evolution of bacterial features' knowledge as well as technologies, instruments, and data analysis tools. Today, more than ever, the implementation of bacterial typing methods that combine a high reliability and accuracy with a rapid, low-cost, and user-friendly performance is highly desirable, especially for clinical microbiology. FT-IR developments for bacterial discrimination at the infra-species level settled on the identification of bacterial groups previously defined by phenotypic or genotypic typing methods. Therefore, this review provides a brief historical overview of main bacterial strain typing methods, and a comprehensive analysis of the fundamentals and applications of Fourier transform infrared spectroscopy, a phenotypic-based method with potential for routine strain typing. The different studies on FT-IR-based strain typing of diverse Gram-negative and Gram-positive bacterial species are discussed in light of genotypic, phenotypic, and biochemical aspects, in order to definitively give this methodology credit to be widely accepted by microbiologists. Importantly, the discriminatory biochemical fingerprints observed on FT-IR spectra have been consistently correlated with sugar-based coating structures that besides reflecting strain variation are also of high relevance for the specificity in pathogen-host interactions. Thus, FT-IR-based bacterial typing might not only be useful for quick and reliable strain typing but also to help understanding the diversity, evolution, and host adaptation factors of key bacterial pathogens or subpopulations.

Beyond health: medicines, food supplements, energetics and the commodification of self-performance in Maputo.

With an increasing range of products in global and local markets, more options are available for individuals to enhance their image and their (cognitive, social and physical) performance. These 'performance consumptions' relate to ideals of well-being and improvement, and are based on constructed desires, expectations and needs that go beyond the (often blurred) dichotomy of health and illness. Drawing from mixed-methods research in Maputo, Mozambique, this paper discusses individuals' use of medicines and other substances - pharmaceuticals, food supplements, traditional herbs, cosmetics and energy drinks - for managing different aspects of their everyday lives. Through an overview of the main consumption practices, we explore the underlying purposes and strategies of users, and the perceived legitimacy and risks involved when using a variety of products accessible through formal and informal exchange channels. From tiredness to sexual and aesthetic management, we show how the body becomes the locus of experimentation and investment to perform in accordance with socially expected roles, individual aspirations and everyday tasks. With insights from individuals' accounts in Maputo, we aim to add to discussions on pharmaceuticalisation of body management by showing how the emergence of new performance consumptions is articulated with the reconfiguration of more 'traditional' consumption practices.

Energy balance and CO2 emission in mechanized biomass harvesting in pine stands under thinning.

Biomass is an important component of the Brazilian energy matrix, with a potential contribution of co-products from thinned forests. The aim of this work was to evaluate the energy balance and CO2 potential emission in mechanized biomass harvesting operations in Pinus taeda stands at 9 and 10 years-old and under thinning, searching to support the use of co-product biomass from thinning as a renewable energy source. Thinning was carried out through cut-to-length harvesting method, in which large logs for sawmill and small logs for energy were produced. In addition, tops, needles, barks, and branches were considered as co-products. The balance between consumed energy and emitted CO2 by machines for thinning in relationship to the energy and CO2 in thinned biomass was estimated. Thus, dry matter, energy potential, and CO2 potential emission were evaluated and compared considering thinning stand ages as treatments. Mechanized thinning consumes a large energy and produces CO2, however, the energy consumed by machines is lower than 1% of the estimated energy potential in thinned biomass, while the CO2 emission is lower than 0.5% of the biomass. Therefore, the use of co-product biomass of thinning is an important way to mitigate greenhouse gas emission.

Modeling residual biomass from mechanized wood harvesting with data measured by forest harvester.

The lack of accurate models for estimating residual biomass in wood harvesting operations results in underutilization of this co-product by forestry companies. Due to the lack of this information, forestry operations planning, such as chipping and transport logistics, are influenced, with a consequent increase in costs. Thereby, the aim of this study was to propose and evaluate statistical models to estimate residual biomass of Eucalyptus sp. in wood harvesting operations by means of tree variables measured from harvester processing head. Generalized linear models were composed through stepwise procedure for estimating residual biomass by tree covariates: diameter at breast height, commercial height, commercial limit diameter, and stem commercial volume, considering also their transformations and combinations. Residual biomass distributions with positive skew support the application of generalized linear model and Gamma distribution in random component, since normality assumption in traditional linear regression was a requirement not satisfied in this study. By stepwise procedure, tree variables associated to forest biomass were selected, whose linear combinations resulted in models with high statistical efficiency and accuracy. Thus, models developed in this study are innovative tools to estimate residual biomass in mechanized wood harvesting, in which can be inserted into harvester's hardware to provide real-time information.

Menstrual cycle alters training strain, monotony, and technical training length in young.

The menstrual cycle (MC) phases carry to several psychophysiological alterations; however, no study has investigated the impact of MC phases on training load or technical training. In the present study, we investigated the effect of the follicular phase (FP), ovulatory phase (OP), and luteal phase (LP) on training load and technical training in young athletes. Twelve female athletes performed regular daily training sessions with the rating of perceived exertion (RPE) and duration being registered every session. Training impulse (TRIMP), monotony and strain were calculated. MC symptoms, RPE, and duration were also measured during technical training, which was carried out on a specific day during each phase. The TRIMP was not affected by MC phases during regular training (p > .05), but training monotony and strain were higher in FP compared to OP (p < .05). During the technical training, MC phases did not affect RPE (p > .05), but the session was longer in both FP and LP, compared to OP (p < .05). MC symptoms were exacerbated in FP compared to both OP and LP (p < .05). These findings suggest that MC disorders were elevated during FP, which indicate that monitoring MC phases might provide important feedbacks for programming training and expected results during competitions.

mcr-1 in Carbapenemase-Producing Klebsiella pneumoniae with Hospitalized Patients, Portugal, 2016-2017.

We describe a hospital-based outbreak caused by multidrug-resistant, Klebsiella pneumoniae carbapenemase 3-producing, mcr-1-positive K. pneumoniae sequence type 45 in Portugal. mcr-1 was located in an IncX4 plasmid. Our data highlight the urgent need for systematic surveillance of mcr-1 to support adequate therapeutic choices in the nosocomial setting.

Family dynamics in transthyretin-related familial amyloid polyneuropathy Val30Met: Does genetic risk affect family functioning?

Adult-onset, chronic, genetic diseases like transthyretin-related familial amyloid polyneuropathy Val30Met (TTR-FAP Val30Met), have a major psychosocial impact not only on patients, but also on families. Genetic risk may therefore be an increased factor in psychosocial impact of the disease on these families' functioning. To evaluate impact of genetic risk, a study was conducted to perceive the impact of the illness on families' functioning. Groups of TTR-FAP Val30Met patients, pre-symptomatic carriers, partners and patients with multiple sclerosis (MS), a non-hereditary disease, were studied. Sample included 190 adults: 87 patients and 28 pre-symptomatic carriers for TTR-FAP Val30Met, 41 partners and 34 patients with MS. Family Adaptability and Cohesion Scale IV (FACES IV) and a social-demographic questionnaire were applied. No significant differences were observed between patients and pre-symptomatic carriers and both these and their partners regarding cohesion and flexibility. MS patients scored significantly higher in median scores for balanced scales. Satisfaction and communication levels were also lower in patients with TTR-FAP Val30Met than with MS. Family functioning was perceived as balanced by most TTR-FAP Val30Met patients and pre-symptomatic carriers. These families may be considered as mostly healthy. Difficulties in family communication should be taken into account when caring for these families.

In Endothelial Cells, the Activation or Stimulation of Soluble Guanylyl Cyclase Induces the Nitric Oxide Production by a Mechanism Dependent of Nitric Oxide Synthase Activation.

PURPOSE: In endothelial cells, investigate if the soluble guanylate cyclase (sGC) activation or stimulation is able to potentiate the relaxation in vessels. METHODS: Aortic and coronary rings with and without endothelium were placed in a myograph and cumulative concentration-effect curves for DETA-NO or ataciguat were performed. Nitric oxide (NO) were measured by fluorescence or by selective electrode in human umbilical endothelial cells (HUVECs) in response to some treatments, including ataciguat, 8-Br-cGMP and A23187. RESULTS: The presence of the endothelium potentiated the relaxation induced by DETA-NO in aortic and coronary rings. In addition, in aortic rings the endothelium potentiated the relaxation induced by ataciguat. In the presence of nitric oxide synthase (NOS) inhibitor, the endothelium effect was abolished to DETA-NO or ataciguat, in both vessels. Ataciguat, 8-Br-cGMP and A23187 were able to induce NO production in HUVECs cells. In the presence of NOS inhibitor, the NO production induced by ataciguat and 8-Br-cGMP was abolished. CONCLUSIONS: Our results suggest that in aortic and coronary rings the endothelium potentiates the relaxation induced by activation or stimulation of sGC through a mechanism dependent of NOS activation. This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.

Contributions to the design of rainwater harvesting systems in buildings with green roofs in a Mediterranean climate.

Green roofs (GRs) are becoming a trend in urban areas, favouring thermal performance of buildings, promoting removal of atmospheric pollutants, and acting as possible water collection spots. Rainwater harvesting systems in buildings can also contribute to the management of stormwater runoff reducing flood peaks. These technologies should be enhanced in Mediterranean countries where water scarcity is increasing and the occurrence of extreme events is becoming very significant, as a result of climate change. An extensive pilot GR with three aromatic plant species, Satureja montana, Thymus caespititius and Thymus pseudolanuginosus, designed to study several parameters affecting rainwater runoff, has been in operation for 12 months. Physico-chemical analyses of roof water runoff (turbidity, pH, conductivity, NH4(+), NO3(-), PO4(3-), chemical oxygen demand) have shown that water was of sufficient quality for non-potable uses in buildings, such as toilet flushing. An innovative approach allowed for the development of an expression to predict a 'monthly runoff coefficient' of the GR system. This parameter is essential when planning and designing GRs combined with rainwater harvesting systems in a Mediterranean climate. This study is a contribution to improving the basis for the design of rainwater harvesting systems in buildings with extensive GRs under a Mediterranean climate.