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Mexico’s cheeseartisanal production is similar or higher than industrial production, approximately 100 L cow’s milk are used to made 10 Kg of cheese and 90 L of whey, that could be considered like an environmentally harmful by product because it has a high content of organic matter that promotes the growth and development of Enterococcus. In this study, Enterococcusstrains were identified in whey from four locations of Puebla. Bacterial gram positive were isolated, furthermore their metabolism shown negative catalase, growth at different temperature (10, 40 and 45 °C), pH values (9.2 and 9.6) and NaCl concentrations (4 and 6.5%). Endpoint PCR tests were made to confirm detection of Enterococcus faecium(19 strains) and Enterococcus faecalis(six strains) in whey from three locations, these results suggest that a better sanitary control of whey is necessary to avoid the proliferation of potentially pathogenic microorganisms., La producción de queso artesanal en México iguala o supera a la producción formal, aproximadamente por cada 100 L de leche de bovino se obtienen 10 kg de queso y 90 L de lactosuero, este residuo es considerado como contaminante por su alta carga orgánica que además favorece el desarrollo de Enterococcus. En este trabajo se identificaron Enterococcus en muestras de lactosuero de cuatro localidades en Puebla. Se aislaron bacterias Gram positivas, catalasas negativas, con capacidad para crecer a diferentes temperaturas (10, 40 y 45 °C), valores de pH (9.2 y 9.6), y concentración de NaCl (4 y 6.5%). Se hicieron pruebas de PCR de punto final para confirmar la detección de Enterococcus faecium (19 cepas) y Enterococcus faecalis (seis cepas) en lactosuero de tres localidades, estos resultados sugieren que es necesario una mejor vigilancia sanitaria del lactosuero, para evitar la proliferación de microorganismos potencialmente patógenos.

The emergence of bloodstream infections (BSI) caused by vancomycin-resistant Enterococci (VRE) has caused concern.Nonetheless, it remains unclear whether these types are associated with an excess risk of severe outcomes whencompared with infections caused by vancomycin-susceptible Enterococci (VSE). This cohort study includedhospitalized patients in Denmark with Enterococcus faecium-positive blood cultures collected between 2010 and 2019identified in the Danish Microbiology Database. We estimated 30-day hazard ratio (HR) of death or discharge amongVRE compared to VSE patients adjusted for age, sex, and comorbidity. The cohort included 6071 patients withE. faecium BSI (335 VRE, 5736 VSE) among whom VRE increased (2010–13, 2.6%; 2014–16, 6.3%; 2017–19; 9.4%).Mortality (HR 1.08, 95%CI 0.90–1.29; 126 VRE, 37.6%; 2223 VSE, 37.0%) or discharge (HR 0.89, 95%CI 0.75–1.06; 126VRE, 37.6%; 2386 VSE, 41.6%) was not different between VRE and VSE except in 2014 (HR 1.87, 95% CI 1.18–2.96).There was no interaction between time from admission to BSI (1–2, 3–14, and >14 days) and HR of death (P = 0.14) ordischarge (P = 0.45) after VRE compared to VSE, despite longer time for VRE patients (17 vs. 10 days for VSE, P <0.0001). In conclusion, VRE BSI was not associated with excess morbidity and mortality. The excess mortality in 2014only may be attributed to improved diagnostic- and patient-management practices after 2014, reducing time toappropriate antibiotic therapy. The high level of mortality after E. faecium BSI warrants further study., The emergence of bloodstream infections (BSI) caused by vancomycin-resistant Enterococci (VRE) has caused concern. Nonetheless, it remains unclear whether these types are associated with an excess risk of severe outcomes when compared with infections caused by vancomycin-susceptible Enterococci (VSE). This cohort study included hospitalized patients in Denmark with Enterococcus faecium-positive blood cultures collected between 2010 and 2019 identified in the Danish Microbiology Database. We estimated 30-day hazard ratio (HR) of death or discharge among VRE compared to VSE patients adjusted for age, sex, and comorbidity. The cohort included 6071 patients with E. faecium BSI (335 VRE, 5736 VSE) among whom VRE increased (2010–13, 2.6%; 2014–16, 6.3%; 2017–19; 9.4%). Mortality (HR 1.08, 95%CI 0.90–1.29; 126 VRE, 37.6%; 2223 VSE, 37.0%) or discharge (HR 0.89, 95%CI 0.75–1.06; 126 VRE, 37.6%; 2386 VSE, 41.6%) was not different between VRE and VSE except in 2014 (HR 1.87, 95% CI 1.18–2.96). There was no interaction between time from admission to BSI (1–2, 3–14, and >14 days) and HR of death (P = 0.14) or discharge (P = 0.45) after VRE compared to VSE, despite longer time for VRE patients (17 vs. 10 days for VSE, P < 0.0001). In conclusion, VRE BSI was not associated with excess morbidity and mortality. The excess mortality in 2014 only may be attributed to improved diagnostic- and patient-management practices after 2014, reducing time to appropriate antibiotic therapy. The high level of mortality after E. faecium BSI warrants further study.

The emergence of bloodstream infections (BSI) caused by vancomycin-resistant Enterococci (VRE) has caused concern.Nonetheless, it remains unclear whether these types are associated with an excess risk of severe outcomes whencompared with infections caused by vancomycin-susceptible Enterococci (VSE). This cohort study includedhospitalized patients in Denmark with Enterococcus faecium-positive blood cultures collected between 2010 and 2019identified in the Danish Microbiology Database. We estimated 30-day hazard ratio (HR) of death or discharge amongVRE compared to VSE patients adjusted for age, sex, and comorbidity. The cohort included 6071 patients withE. faecium BSI (335 VRE, 5736 VSE) among whom VRE increased (2010–13, 2.6%; 2014–16, 6.3%; 2017–19; 9.4%).Mortality (HR 1.08, 95%CI 0.90–1.29; 126 VRE, 37.6%; 2223 VSE, 37.0%) or discharge (HR 0.89, 95%CI 0.75–1.06; 126VRE, 37.6%; 2386 VSE, 41.6%) was not different between VRE and VSE except in 2014 (HR 1.87, 95% CI 1.18–2.96).There was no interaction between time from admission to BSI (1–2, 3–14, and >14 days) and HR of death (P = 0.14) ordischarge (P = 0.45) after VRE compared to VSE, despite longer time for VRE patients (17 vs. 10 days for VSE, P <0.0001). In conclusion, VRE BSI was not associated with excess morbidity and mortality. The excess mortality in 2014only may be attributed to improved diagnostic- and patient-management practices after 2014, reducing time toappropriate antibiotic therapy. The high level of mortality after E. faecium BSI warrants further study., The emergence of bloodstream infections (BSI) caused by vancomycin-resistant Enterococci (VRE) has caused concern. Nonetheless, it remains unclear whether these types are associated with an excess risk of severe outcomes when compared with infections caused by vancomycin-susceptible Enterococci (VSE). This cohort study included hospitalized patients in Denmark with Enterococcus faecium-positive blood cultures collected between 2010 and 2019 identified in the Danish Microbiology Database. We estimated 30-day hazard ratio (HR) of death or discharge among VRE compared to VSE patients adjusted for age, sex, and comorbidity. The cohort included 6071 patients with E. faecium BSI (335 VRE, 5736 VSE) among whom VRE increased (2010–13, 2.6%; 2014–16, 6.3%; 2017–19; 9.4%). Mortality (HR 1.08, 95%CI 0.90–1.29; 126 VRE, 37.6%; 2223 VSE, 37.0%) or discharge (HR 0.89, 95%CI 0.75–1.06; 126 VRE, 37.6%; 2386 VSE, 41.6%) was not different between VRE and VSE except in 2014 (HR 1.87, 95% CI 1.18–2.96). There was no interaction between time from admission to BSI (1–2, 3–14, and >14 days) and HR of death (P = 0.14) or discharge (P = 0.45) after VRE compared to VSE, despite longer time for VRE patients (17 vs. 10 days for VSE, P < 0.0001). In conclusion, VRE BSI was not associated with excess morbidity and mortality. The excess mortality in 2014 only may be attributed to improved diagnostic- and patient-management practices after 2014, reducing time to appropriate antibiotic therapy. The high level of mortality after E. faecium BSI warrants further study.

Bacteriocins produced by enterococci, referred to as enterocins, possess great interest for their potential use as biopreservatives in food and feed, as well as alternative antimicrobials in humans and animals. In this context, the aim of the present study was to determine the antimicrobial activity and the presence of bacteriocin structural genes in fecal enterococcal isolates from animal origins. Evaluation of the direct antimicrobial activity of 253 isolates from wild boars (Sus scrofa, n = 69), mullets (Liza ramada, n = 117), and partridges (Perdix perdix, n = 67) against eight indicator bacterial strains (including Listeria monocytogenes, Pediococcus pentosaceus, and Enterococcus spp.) showed that 177 (70%) exerted antimicrobial activity against at least one indicator microorganism. From these isolates, 123 were further selected on the basis of their inhibition group, and 81 were found to be producers of bacteriocins active against Listeria monocytogenes. Analysis of the presence of enterocin structural genes in a subset of 36 isolates showed that 70% harbored one or more of the evaluated genes, those of enterocin P and hiracin JM79 being the most prevalent. These results show that wild animals constitute an appropriate source for the isolation of bacteriocinogenic enterococci., Ministerio de Ciencia e Innovación, Ministerio de Educación, Cultura y Deporte, Ministerio de Ciencia y Tecnología, Comunidad de Madrid, European Commission and Agência Nacional from Portugal, Sección Dptal. de Nutrición y Ciencia de los Alimentos (Veterinaria), Fac. de Veterinaria, TRUE, pub

Antimicrobial resistance is considered a One Health problem, affecting human, livestock, and environmental health. The overuse of antimicrobials in human and livestock settings exerts selective pressure, resulting in the proliferation of antimicrobial-resistant bacterial strains. Enterococci and Escherichia coli exist ubiquitously in One Health sectors and are therefore used as indicator bacteria in antimicrobial resistance surveillance programs. These bacteria carry antimicrobial resistance genes on mobile genetic elements that may transfer to other bacterial species, most importantly to pathogenic bacteria. In this thesis, we characterized Enterococcus species recovered from One Health sectors and carbapenem-resistant bacteria in Canada. Phenotypic characterization was conducted using the disc diffusion method according to standard CLSI protocols, and whole sequencing using short and long-read platforms was used for genotypic characterization. Additionally, whole-genome comparative genomics was used to investigate the genomic relatedness among isolates from One Health sectors and the effect of antimicrobial usage on antimicrobial resistance in each sector. We identified Enterococcus hirae as the prevalent Enterococcus spp. in bovine and swine production systems, while Enterococcus faecium was predominantly found in poultry. In human clinical isolates, Enterococcus faecium and Enterococcus faecalis were predominantly identified. In antimicrobial resistance profiling of Enterococcus spp., we found that tetracycline (tetL, tetM) and macrolide resistance (ermB) genes exist universally in E. faecium and E. faecalis from all One Health sectors. However, these resistance genes were predominantly found in bovine-sourced E. hirae compared to human settings. Across livestock sectors, the analysis showed that multidrug-resistant E. faecium and E. faecalis isolates were more commonly found in poultry than in swine and bovine cattle. The reconstruction of a core-genome-based p

This study aimed to explore the probiogenomic characteristics of artisanal bacteriocin-producing Enterococcus faecium BGZLM1-5 and its potential application in reducing Listeria monocytogenes in a milk model. The BGZLM1-5 strain was isolated from raw cow’s milk from households in the Zlatar Mountain region. The whole genome sequencing approach and bioinformatics analyses reveal that the strain BGZLM1-5 is non-pathogenic to humans. Bacteriocin-containing supernatant was thermally stable and antimicrobial activity retained 75% of the initial activity compared with that of the control after treatment at 90 °C for 30 min. Antimicrobial activity maintained relative stability at pH 3–11 and retained 62.5% of the initial activity compared with that of the control after treatment at pH 1, 2, and 12. The highest activity of the partially purified bacteriocin was obtained after precipitation at 40% saturation with ammonium sulfate and further purification by mixing with chloroform. Applying 3% and 5% (v/v) of the bacteriocin-containing supernatant and 0.5% (v/v) of the partially purified bacteriocin decreased the viable number of L. monocytogenes ATCC19111 after three days of milk storage by 23.5%, 63.5%, and 58.9%, respectively.

One of the major genera of the lactic acid bacteria family, Enterococcus sp., has a controversial status, reflected in the fact that enterococci are utilized as starter cultures and probiotics, in addition to being known to cause nosocomial infections. The qualified presumption of the safety list and the widely acknowledged safe status for Enterococcus species are absent. Rich sources of Enterococcus faecium species with possible probiotic characteristics can be found in artisanal dairy products, typically made from raw milk. To further understand the probiotic potential and health-promoting effects, this study looked at the presence of virulence factors and adhesion properties of En. faecium isolated from artisanal dairy products from Western Balkan countries.

Smoothie drinks are currently very popular drinks sold especially in fast food establishments. However, smoothies are a significant source of microorganisms. The aim of this study was to evaluate the microbiological quality of smoothies purchased in Eastern Bohemia. A higher prevalence of mesophilic aerobic bacteria (5.4–7.2 log CFU/mL), yeast (4.4–5.9 log CFU/mL) and coliform bacteria (3.1–6.0 log CFU/mL) was observed in vegetable smoothies, in which even the occurrence of enterococci (1.6–3.3 log CFU/mL) was observed. However, the occurrence of S. aureus, Salmonella spp. and Listeria spp. was not observed in any samples. Nevertheless, antimicrobial resistance was observed in 71.8% of the isolated strains. The highest level of resistance was found in isolates from smoothie drinks with predominantly vegetable contents (green smoothie drinks). Considerable resistance was observed in Gram-negative rods, especially to amoxicillin (82.2%) and amoxicillin with clavulanic acid (55.6%). Among enterococci, only one vancomycin-resistant strain was detected. The vast majority of isolated strains were able to form biofilms at a significant level, which increases the clinical importance of these microorganisms. The highest biofilm production was found in Pseudomonas aeruginosa, Kocuria kristinae and Klebsiella pneumoniae. Overall, significant biofilm production was also noted among isolates of Candida spp., Smoothie nápoje jsou v současnosti velmi oblíbené nápoje prodávané zejména v provozovnách rychlého občerstvení. Smoothie jsou však také významným zdrojem mikroorganismů. Cílem této studie bylo zhodnotit mikrobiologickou kvalitu smoothie nápojů zakoupených ve východních Čechách. Byla pozorována vyšší prevalence mezofilních aerobních bakterií (5,4–7,2 log CFU/ml), kvasinek (4,4–5,9 log CFU/ml) a koliformních bakterií (3,1–6,0 log CFU/ml) zejména v zeleninových smoothie nápojích, u kterých byl zaznamenán dokonce výskyt enterokoků (1,6–3,3 log CFU/ml). Nicméně výskyt S. aureus, Salmonella spp. a Listeria spp. nebyl pozorován v žádném vzorku. Antimikrobiální rezistence byla pozorována u 71,8 % izolovaných kmenů. Nejvyšší úroveň rezistence byla zjištěna u izolátů ze smoothie nápojů s dominantním obsahem zeleniny (zelené smoothie nápoje). Značná rezistence byla pozorována u gramnegativních tyčinek, zejména vůči amoxicilinu (82,2 %) a amoxicilinu s kyselinou klavulanovou (55,6 %). Mezi enterokoky byl detekován pouze jeden kmen rezistentní k vankomycinu. Naprostá většina izolovaných kmenů byla schopna tvořit biofilmy, a to na významné úrovni, což zvyšuje klinický význam těchto mikroorganismů. Nejvyšší tvorba biofilmu byla zjištěna u Pseudomonas aeruginosa, Kocuria kristinae a Klebsiella pneumoniae. Celkově byla významná produkce biofilmu zaznamenána také mezi izoláty Candida spp.

Antimicrobial resistance is considered a One Health problem, affecting human, livestock, and environmental health. The overuse of antimicrobials in human and livestock settings exerts selective pressure, resulting in the proliferation of antimicrobial-resistant bacterial strains. Enterococci and Escherichia coli exist ubiquitously in One Health sectors and are therefore used as indicator bacteria in antimicrobial resistance surveillance programs. These bacteria carry antimicrobial resistance genes on mobile genetic elements that may transfer to other bacterial species, most importantly to pathogenic bacteria. In this thesis, we characterized Enterococcus species recovered from One Health sectors and carbapenem-resistant bacteria in Canada. Phenotypic characterization was conducted using the disc diffusion method according to standard CLSI protocols, and whole sequencing using short and long-read platforms was used for genotypic characterization. Additionally, whole-genome comparative genomics was used to investigate the genomic relatedness among isolates from One Health sectors and the effect of antimicrobial usage on antimicrobial resistance in each sector. We identified Enterococcus hirae as the prevalent Enterococcus spp. in bovine and swine production systems, while Enterococcus faecium was predominantly found in poultry. In human clinical isolates, Enterococcus faecium and Enterococcus faecalis were predominantly identified. In antimicrobial resistance profiling of Enterococcus spp., we found that tetracycline (tetL, tetM) and macrolide resistance (ermB) genes exist universally in E. faecium and E. faecalis from all One Health sectors. However, these resistance genes were predominantly found in bovine-sourced E. hirae compared to human settings. Across livestock sectors, the analysis showed that multidrug-resistant E. faecium and E. faecalis isolates were more commonly found in poultry than in swine and bovine cattle. The reconstruction of a core-genome-based p

Antimicrobial resistance is considered a One Health problem, affecting human, livestock, and environmental health. The overuse of antimicrobials in human and livestock settings exerts selective pressure, resulting in the proliferation of antimicrobial-resistant bacterial strains. Enterococci and Escherichia coli exist ubiquitously in One Health sectors and are therefore used as indicator bacteria in antimicrobial resistance surveillance programs. These bacteria carry antimicrobial resistance genes on mobile genetic elements that may transfer to other bacterial species, most importantly to pathogenic bacteria. In this thesis, we characterized Enterococcus species recovered from One Health sectors and carbapenem-resistant bacteria in Canada. Phenotypic characterization was conducted using the disc diffusion method according to standard CLSI protocols, and whole sequencing using short and long-read platforms was used for genotypic characterization. Additionally, whole-genome comparative genomics was used to investigate the genomic relatedness among isolates from One Health sectors and the effect of antimicrobial usage on antimicrobial resistance in each sector. We identified Enterococcus hirae as the prevalent Enterococcus spp. in bovine and swine production systems, while Enterococcus faecium was predominantly found in poultry. In human clinical isolates, Enterococcus faecium and Enterococcus faecalis were predominantly identified. In antimicrobial resistance profiling of Enterococcus spp., we found that tetracycline (tetL, tetM) and macrolide resistance (ermB) genes exist universally in E. faecium and E. faecalis from all One Health sectors. However, these resistance genes were predominantly found in bovine-sourced E. hirae compared to human settings. Across livestock sectors, the analysis showed that multidrug-resistant E. faecium and E. faecalis isolates were more commonly found in poultry than in swine and bovine cattle. The reconstruction of a core-genome-based p

Antimicrobial resistance is considered a One Health problem, affecting human, livestock, and environmental health. The overuse of antimicrobials in human and livestock settings exerts selective pressure, resulting in the proliferation of antimicrobial-resistant bacterial strains. Enterococci and Escherichia coli exist ubiquitously in One Health sectors and are therefore used as indicator bacteria in antimicrobial resistance surveillance programs. These bacteria carry antimicrobial resistance genes on mobile genetic elements that may transfer to other bacterial species, most importantly to pathogenic bacteria. In this thesis, we characterized Enterococcus species recovered from One Health sectors and carbapenem-resistant bacteria in Canada. Phenotypic characterization was conducted using the disc diffusion method according to standard CLSI protocols, and whole sequencing using short and long-read platforms was used for genotypic characterization. Additionally, whole-genome comparative genomics was used to investigate the genomic relatedness among isolates from One Health sectors and the effect of antimicrobial usage on antimicrobial resistance in each sector. We identified Enterococcus hirae as the prevalent Enterococcus spp. in bovine and swine production systems, while Enterococcus faecium was predominantly found in poultry. In human clinical isolates, Enterococcus faecium and Enterococcus faecalis were predominantly identified. In antimicrobial resistance profiling of Enterococcus spp., we found that tetracycline (tetL, tetM) and macrolide resistance (ermB) genes exist universally in E. faecium and E. faecalis from all One Health sectors. However, these resistance genes were predominantly found in bovine-sourced E. hirae compared to human settings. Across livestock sectors, the analysis showed that multidrug-resistant E. faecium and E. faecalis isolates were more commonly found in poultry than in swine and bovine cattle. The reconstruction of a core-genome-based p

Vancomycin-resistant Enterococcus faecium represents a health threat due to its ability to spread and cause outbreaks. MALDI-TOF MS has demonstrated its usefulness for E. faecium identification, but its implementation for antimicrobial resistance detection is still under evaluation. This study assesses the repeatability of MALDI-TOF MS for peak analysis and its performance in the discrimination of vancomycin-susceptible (VSE) from vancomycin-resistant isolates (VRE). The study was carried out on protein spectra from 178 E. faecium unique clinical isolates—92 VSE, 31 VanA VRE, 55 VanB VRE-, processed with Clover MS Data Analysis software. Technical and biological repeatability were assayed. Unsupervised (principal component analysis, (PCA)) and supervised algorithms (support vector machine (SVM), random forest (RF) and partial least squares–discriminant analysis (PLS-DA)) were applied. The repeatability assay was performed with 18 peaks common to VSE and VRE with intensities above 1.0% of the maximum peak intensity. It showed lower variability for normalized data and for the peaks within the 3000–9000 m/z range. It was found that 80.9%, 79.2% and 77.5% VSE vs. VRE discrimination was achieved by applying SVM, RF and PLS-DA, respectively. Correct internal differentiation of VanA from VanB VRE isolates was obtained by SVM in 86.6% cases. The implementation of MALDI-TOF MS and peak analysis could represent a rapid and effective tool for VRE screening. However, further improvements are needed to increase the accuracy of this approach., Instituto de Salud Carlos III (ISCIII)/ FEDER, Depto. de Medicina, Fac. de Medicina, TRUE, pub

Introduction: We previously demonstrated the efficacy of a frozen dalbavancin–heparin (DH) lock solution against biofilms of staphylococci. However, as enterococci also commonly cause catheter-related bloodstream infections (CRBSI), we assessed the bioactivity of frozen dalbavancin (D) and DH against enterococci. Methods: Over 6 months, we compared the bioactivity of a solution of DH (1 mg/ml) with that of D in terms of cfu counts and metabolic activity against biofilms of Enterococcus faecalis and Enterococcus faecium (four strains each). For each solution, we individually compared results obtained at each time point (months 3 and 6) with baseline (month 0). We also compared the median DH value of each variable at baseline and at months 3 and 6 of freezing with the values obtained for D alone. We used both statistical and clinical criteria when results were within 25% of the reference value. Results: At the end of the experiment (month 6), neither a statistically nor a clinically significant reduction in the bioactivity of D solution was observed in terms of cfu count and metabolic activity against enterococcal biofilms. Regarding the DH solution, we found both statistical and clinical significance in the median percentage reduction in metabolic activity between months 0 and 6 in E. faecalis strains (51.8% vs. 77.8%, P = 0.007). Moreover, after freezing, the DH solution lost significant bioactivity compared with the D solution, especially in E. faecalis. Conclusion: A dalbavancin lock solution can be frozen for up to 6 months with no negative effect on its bioactivity against enterococcal biofilms. However, when combined with heparin, its efficacy was reduced. Therefore, we recommend that if lock therapy with frozen dalbavancin is used in the management of enterococcal C-RBSI, heparin should be added simultaneously at the time of catheter lock., Ministerio de Ciencia, Innovación y Universidades (España)/Instituto de Salud Carlos III, Instituto de Salud Carlos III/Fondo Europeo de Desarrollo Regional, Consejería de Educación, Juventud y Deporte (Comunidad de Madrid)/ Fondo Social Europeo, Depto. de Genética, Fisiología y Microbiología, Depto. de Medicina, Fac. de Ciencias Biológicas, Fac. de Medicina, TRUE, pub

Enterococci represent a significant part of the non-starter LAB microbiota of artisanal cheeses produced mainly from raw milk. Common approaches to safety evaluation of enterococci isolates include assessment of antimicrobial resistance and virulence potential. Hence, a collection of 47 (n = 22, Serbia; n = 25, Slovenia) dairy enterococcal isolates, of which E. faecalis (n = 28), E. faecium (n = 11), E. durans (n = 5), E. casseliflavus (n = 2), and E. gallinarum (n = 1), was analyzed. The susceptibility to 12 antimicrobials was tested using a broth microdilution method, and the presence of the selected antimicrobial resistance and virulence genes was investigated using PCR. Isolates were resistant to tetracycline (TET) (25.5%), erythromycin (ERY) (17.0%), gentamycin and chloramphenicol (CHL) (∼6%). No resistance to ampicillin (AMP), ciprofloxacin (CIP), daptomycin (DAP), linezolid (LZD), teicoplanin (TEI), tigecycline (TGC) and vancomycin (VAN) was detected. Among all the resistance determinants analyzed, ermB gene was detected most frequently. All 10 virulence genes analyzed were detected with a distribution of cpd (72.3%), cob and ccf (70.2%), gelE (68.1%), hyl (59.6%), agg (53.2%) and esp (46.8%). The genes encoding cytolysin (cylA, cylM and cylB) were amplified to a lesser extent (21.3%, 21.3% and 12.8%, respectively). However, due to the limited number of enterococci isolates analyzed in the present study, further studies are still required in order to better document the safety status of dairy enterococci.

Dispersion of transferable oxazolidinone resistance genes among enterococci poses a serious problem to human health. Prompt detection of bacteria carrying these genes is crucial to avoid their spread to multidrug-resistant bacteria. The aim of the study was to describe the presence of optrA-positive isolates among enterococci in a Spanish hospital, and to determine their genetic context and location through whole genome sequencing. All enterococci recovered in a Spanish hospital (Hospital El Bierzo; HEB) from February to December 2018 (n = 443), with minimal inhibitory concentrations (MICs) to linezolid (LZD) ≥4 mg/L, were tested by polymerase chain reaction for the presence of cfr, optrA, and poxtA transferable genes. Only four Enterococcus faecalis isolates (0.9%) had LZD MICs ≥4 mg/L and none of them was positive for cfr or poxtA genes. However, the optrA gene was detected in three isolates collected from urine samples of community patients, whose genomes were sequenced and subjected to bioinformatics analysis. These isolates belonged to different clones: ST7, ST480, and ST585. In these three isolates, the optrA gene was located on plasmids, associated with IS1216 in different arrays. In one isolate, the optrA plasmid coexists with a second plasmid, which carried multiple resistance genes for different classes of antibiotics. Detection of optrA-positive E. faecalis isolates in the community is a matter of concern. The spread of these bacteria into hospital settings, particularly in those, such as the HEB, where vancomycin-resistant enterococci are endemic, should be avoided, to preserve the efficacy of the last-resort oxazolidinones.

Els resultats presentats en l’informe extret de la European Centre for Disease Prevention and Control (ECDC) estan basats en dades de resistència antimicrobiana (RAM) d’aïllats invasius comunicats a la xarxa europea de vigilància de la resistència antimicrobiana (EARS-Net) per 30 països de la Unió Europea (UE) i de l’Espai Econòmic Europeu (EEE) el 2019 (dades referent al 2018), i en anàlisis de tendències de les dades reportades pels països participants per al període 2015-2018. El projecte Surevillance of antimicrobial resistance in Europe 2018 és un estudi que té com a objectiu mostrar la RAM que hi ha a Europa, mostrant les variacions que hi ha entre els diferents països, diferents bacteris i antibiòtics. En aquest estudi, a través de diferents tècniques d’Anàlisis Multivariant, s’analitza la resistència dels diferents tipus de bacteris a la combinació d’antibiòtics que s’ha observat en els diferents països seleccionats i l’evolució que han tingut aquests tractaments.

Aims: To determine inactivation of Cryptosporidium parvum oocysts and reduction of Escherichia coli and enterococci in cattle slurry added aqueous ammonia. Methods and Results: Escherichia coli, enterococci and nonviable C. parvum oocysts (DAPI+PI+) were enumerated every second day for 2 weeks in cattle slurry amended with 60 mmol l−1 aq. ammonia and compared with untreated slurry at three temperatures. Regardless of temperature, the proportion of nonviable C. parvum oocysts increased significantly faster over time in slurry with added ammonia than raw slurry (P = 0·021) corresponding to 62·0% higher inactivation (P = 0·001) at day 14. Additionally, 91·8% fewer E. coli and 27·3% fewer enterococci were observed in slurry added ammonia at day 14 compared to raw slurry. Conclusion: The addition of aqueous ammonia to raw slurry significantly reduced the viability of C. parvum oocysts and numbers of bacterial indicators. Hence, ammonia is usable at lower pathogen concentrations in slurry before application to agricultural land. Significance and Impact of the Study: Livestock waste is a valuable source of plant nutrients and organic matter, but may contain high concentrations of pathogens like E. coli and Cryptosporidium sp. that can be spread in the environment, and cause disease outbreaks. However, die-off rates of pathogens in organic waste can increase following increasing ammonia concentrations.

Aims: To determine inactivation of Cryptosporidium parvum oocysts and reduction of Escherichia coli and enterococci in cattle slurry added aqueous ammonia. Methods and Results: Escherichia coli, enterococci and nonviable C. parvum oocysts (DAPI+PI+) were enumerated every second day for 2 weeks in cattle slurry amended with 60 mmol l−1 aq. ammonia and compared with untreated slurry at three temperatures. Regardless of temperature, the proportion of nonviable C. parvum oocysts increased significantly faster over time in slurry with added ammonia than raw slurry (P = 0·021) corresponding to 62·0% higher inactivation (P = 0·001) at day 14. Additionally, 91·8% fewer E. coli and 27·3% fewer enterococci were observed in slurry added ammonia at day 14 compared to raw slurry. Conclusion: The addition of aqueous ammonia to raw slurry significantly reduced the viability of C. parvum oocysts and numbers of bacterial indicators. Hence, ammonia is usable at lower pathogen concentrations in slurry before application to agricultural land. Significance and Impact of the Study: Livestock waste is a valuable source of plant nutrients and organic matter, but may contain high concentrations of pathogens like E. coli and Cryptosporidium sp. that can be spread in the environment, and cause disease outbreaks. However, die-off rates of pathogens in organic waste can increase following increasing ammonia concentrations.

Aims: To determine inactivation of Cryptosporidium parvum oocysts and reduction of Escherichia coli and enterococci in cattle slurry added aqueous ammonia. Methods and Results: Escherichia coli, enterococci and non-viable C. parvum oocysts (DAPI+ PI+) were enumerated every second day for two weeks in cattle slurry amended with 60 mmol l-1 aq. ammonia and compared with untreated slurry at three temperatures. Regardless of temperature, the proportion of non-viable C. parvum oocysts increased significantly faster over time in slurry with added ammonia than raw slurry (p=0.021) corresponding to 62.0% higher inactivation (P=0.001) at day 14. Additionally, 91.8% fewer E. coli and 27.3% fewer enterococci was observed slurry added ammonia at day 14 compared raw slurry. Conclusion: The addition of aqueous ammonia to raw slurry significantly reduced the viability of C. parvum oocysts and numbers of bacterial indicators. Hence, ammonia is usable to lower pathogen concentrations in slurry prior before application to agricultural land. Significance and Impact of Study: Livestock waste is a valuable source of plant nutrients and organic matter, but may contain high concentrations of pathogens like Escherichia coli and Cryptosporidium spp. that can be spread in the environment, and cause disease outbreaks. However, die-off rates of pathogens in organic waste can increase following increasing ammonia concentrations.

Bacteria transport through soil is a complex process particularly when the cells are released from solid manures and co-transported with particles. This study focuses on understanding of the Escherichia coli release from different particle fractions (0.25-, 0.5-, 1-, and 2-mm) of solid manure and evaluating different influent boundary conditions during cell release from manure and when a solid manure is applied to the soil. The 0.25-mm and 2-mm particle sizes resulted a greater cell release compared to 0.5-mm and 1-mm fractions (p < 0.05). The shape and magnitude of the cell release curves (CRCs) from the original manure bulk were mainly influenced by the two 0.25-mm and 2-mm fractions, respectively. The arithmetic mean for normalizing the CRCs and the time variable- based normalized CRCs for the manure-treated soil were the robust variables in evaluation of the experimental data. However, a single maximum bacteria concentration could provide the realistic dataset for the modeling process. Evaluation of the root-mean-squared-error and Akaike criterion showed that the two- and three-parametric models are recommended for simulating the cell release from solid manure in comparison with one parametric models. This study also suggests considering separate microbial release evaluations, with regards to influent concentration, for manure and manure-treated soils to propose best management practices for controlling bacteria pollution. Further research will reveal the key roles of different woody components and soluble material ratios for the various solid manures in bacteria release.

Biofilters are increasingly being implemented in urban environments to treat stormwater but knowledge on their pathogen removal capacity at field scale is limited. Adequate pathogen removal is, however, needed if treated water is reused. This research evaluates the fate of Escherichia coli, enterococci and Campylobacter in a field scale biofilter in Rotterdam. This biofilter is part of the Urban Waterbuffer Spangen that combines biofiltration with Aquifer Storage and Recovery (ASR) to provide irrigation water for Sparta’s sports field. Microbiological water quality was analysed to assess the removal capacity of the biofilter. To identify factors causing adverse treatment performance, system construction, and operation were investigated, water flows in the biofilter were assessed, and hydraulic conductivity was measured. Lastly, model simulations were used to investigate how short-circuiting and event frequency and duration affect microbiological outflow concentrations. Results showed leaching of enterococci and Campylobacter and minimal retention of Escherichia coli. Leaching is likely caused by secondary contamination with bird faeces and multiple design and operational choices lead to low microbial retention. Coarse filter media results in high hydraulic conductivity, and consequently, short hydraulic retention times. Electrical conductivity measurements revealed short-circuiting pathways between the inlet and outlet. Lastly, uneven distribution of feed water presumably reduced the effective reactor volume. It is recommended to upgrade biofilter design by incorporating small particles in the filter media and maximizing the distance between the inlet and outlet. Additionally, reintroduce a ponding zone and rearranging event frequency and duration to lower microbial outflow loads is advised. Model simulations suggest that short, frequent events are beneficial to control microbial loads. However, further work to clarify microbial decay rates is recommended. To conclu, Civil Engineering | Environmental Engineering

Biofilters are increasingly being implemented in urban environments to treat stormwater but knowledge on their pathogen removal capacity at field scale is limited. Adequate pathogen removal is, however, needed if treated water is reused. This research evaluates the fate of Escherichia coli, enterococci and Campylobacter in a field scale biofilter in Rotterdam. This biofilter is part of the Urban Waterbuffer Spangen that combines biofiltration with Aquifer Storage and Recovery (ASR) to provide irrigation water for Sparta’s sports field. Microbiological water quality was analysed to assess the removal capacity of the biofilter. To identify factors causing adverse treatment performance, system construction, and operation were investigated, water flows in the biofilter were assessed, and hydraulic conductivity was measured. Lastly, model simulations were used to investigate how short-circuiting and event frequency and duration affect microbiological outflow concentrations. Results showed leaching of enterococci and Campylobacter and minimal retention of Escherichia coli. Leaching is likely caused by secondary contamination with bird faeces and multiple design and operational choices lead to low microbial retention. Coarse filter media results in high hydraulic conductivity, and consequently, short hydraulic retention times. Electrical conductivity measurements revealed short-circuiting pathways between the inlet and outlet. Lastly, uneven distribution of feed water presumably reduced the effective reactor volume. It is recommended to upgrade biofilter design by incorporating small particles in the filter media and maximizing the distance between the inlet and outlet. Additionally, reintroduce a ponding zone and rearranging event frequency and duration to lower microbial outflow loads is advised. Model simulations suggest that short, frequent events are beneficial to control microbial loads. However, further work to clarify microbial decay rates is recommended. To conclu, Civil Engineering | Environmental Engineering

[Background] Enterococcal endocarditis (EE) is a growing entity in Western countries. However, quality data from large studies is lacking., [Objectives] The purpose of this study was to describe the characteristics and analyze the prognostic factors of EE in the GAMES cohort., [Methods] This was a post hoc analysis of a prospectively collected cohort of patients from 35 Spanish centers from 2008 to 2016. Characteristics and outcomes of 516 cases of EE were compared with those of 3,308 cases of nonenterococcal endocarditis (NEE). Logistic regression and Cox proportional hazards regression analysis were performed to investigate risk factors for in-hospital and 1-year mortality, as well as relapses., [Results] Patients with EE were significantly older; more frequently presented chronic lung disease, chronic heart failure, prior endocarditis, and degenerative valve disease; and had higher median age-adjusted Charlson score. EE more frequently involved the aortic valve and prosthesis (64.3% vs. 46.7%; p < 0.001; and 35.9% vs. 28.9%; p = 0.002, respectively) but less frequently pacemakers/defibrillators (1.5% vs. 10.5%; p < 0.001), and showed higher rates of acute heart failure (45% vs. 38.3%; p = 0.005). Cardiac surgery was less frequently performed in EE (40.7% vs. 45.9%; p = 0.024). No differences in in-hospital and 1-year mortality were found, whereas relapses were significantly higher in EE (3.5% vs. 1.7%; p = 0.035). Increasing Charlson score, LogEuroSCORE, acute heart failure, septic shock, and paravalvular complications were risk factors for mortality, whereas prior endocarditis was protective and persistent bacteremia constituted the sole risk factor for relapse., [Conclusions] Besides other baseline and clinical differences, EE more frequently affects prosthetic valves and less frequently pacemakers/defibrillators. EE presents higher rates of relapse than NEE.

[EN] Properties of enterococci isolated from the Hungarian breed Pannon White were studied to spread knowledge regarding the properties of microbiota in rabbits from the basic research standpoint and to select a beneficial candidate for application in husbandry. Faeces from 113 Pannon White rabbits (mostly maternal line and some paternal line) were collected. They were sampled from rabbits varying in age and sex (82 kits, 6 does, 6 bucks, 19 adult rabbits), which were aged 2 wk (14 suckling rabbits), 6 and 8 wk (68 weaning and post-weaning rabbits), adult rabbits (31, one year). Faecal mixtures were sampled into sterile packs with faeces from 5-6 animals in each. The total count of enterococci from these Pannon White rabbits reached, on av. 5.28±0.29 colony forming units/g (log10). Among the 19 enterococci, 14 E. faecalis and 5 E. faecium were detected using 3 identification methods. The enterococci were mostly resistant to antibiotics. They were non-haemolytic, Dnase and urease negative. They did not form biofilm. They were free of the hylEfm gene and IS16 genes; the most frequently detected genes were gelE, efaAfm, efaAfs. Based on these results, E. faecium EF9a was selected for further analysis.

[EN] Properties of enterococci isolated from the Hungarian breed Pannon White were studied to spread knowledge regarding the properties of microbiota in rabbits from the basic research standpoint and to select a beneficial candidate for application in husbandry. Faeces from 113 Pannon White rabbits (mostly maternal line and some paternal line) were collected. They were sampled from rabbits varying in age and sex (82 kits, 6 does, 6 bucks, 19 adult rabbits), which were aged 2 wk (14 suckling rabbits), 6 and 8 wk (68 weaning and post-weaning rabbits), adult rabbits (31, one year). Faecal mixtures were sampled into sterile packs with faeces from 5-6 animals in each. The total count of enterococci from these Pannon White rabbits reached, on av. 5.28±0.29 colony forming units/g (log10). Among the 19 enterococci, 14 E. faecalis and 5 E. faecium were detected using 3 identification methods. The enterococci were mostly resistant to antibiotics. They were non-haemolytic, Dnase and urease negative. They did not form biofilm. They were free of the hylEfm gene and IS16 genes; the most frequently detected genes were gelE, efaAfm, efaAfs. Based on these results, E. faecium EF9a was selected for further analysis.

We describe clonal shifts in vanA Enterococcus faecium isolates from clinical samples obtained from patients in Denmark from 2015 to the first quarter (Q1) of 2019. During Q1 2019, the vancomycin-variable enterococci (VVE) ST1421-CT1134 vanA E. faecium became the most dominant vanA E. faecium clone and has spread to all five regions in Denmark. Among 174 E. faecium isolates with vanA, vanB or vanA/vanB genes in Q1 2019, 44% belonged to this type.

The SuperLinezolid medium was developed for screening resistance to linezolid (LZD) in Gram-positive bacteria (Staphylococcus spp., Enterococcus spp.). It was evaluated using LZD-susceptible (n = 20) and LZD-resistant (n = 17) Gram-positive isolates. The sensitivity was found to be 82% at 24 h (3 out of 17 isolates being missed), and reached 100% at 48 h. At 48 h, a single LZD-susceptible isolate grew (specificity 95%). By testing stools spiked with LZD-resistant Gram-positive strains, an excellent performance of the medium was observed, with a lowest detection limit ranging from 101 to 102 CFU/ml. Overall, this medium is accurate for detection of LZD-resistant Gram-positive isolates after 24 h of culture.

[EN] Properties of enterococci isolated from the Hungarian breed Pannon White were studied to spread knowledge regarding the properties of microbiota in rabbits from the basic research standpoint and to select a beneficial candidate for application in husbandry. Faeces from 113 Pannon White rabbits (mostly maternal line and some paternal line) were collected. They were sampled from rabbits varying in age and sex (82 kits, 6 does, 6 bucks, 19 adult rabbits), which were aged 2 wk (14 suckling rabbits), 6 and 8 wk (68 weaning and post-weaning rabbits), adult rabbits (31, one year). Faecal mixtures were sampled into sterile packs with faeces from 5-6 animals in each. The total count of enterococci from these Pannon White rabbits reached, on av. 5.28±0.29 colony forming units/g (log10). Among the 19 enterococci, 14 E. faecalis and 5 E. faecium were detected using 3 identification methods. The enterococci were mostly resistant to antibiotics. They were non-haemolytic, Dnase and urease negative. They did not form biofilm. They were free of the hylEfm gene and IS16 genes; the most frequently detected genes were gelE, efaAfm, efaAfs. Based on these results, E. faecium EF9a was selected for further analysis.

[EN] Properties of enterococci isolated from the Hungarian breed Pannon White were studied to spread knowledge regarding the properties of microbiota in rabbits from the basic research standpoint and to select a beneficial candidate for application in husbandry. Faeces from 113 Pannon White rabbits (mostly maternal line and some paternal line) were collected. They were sampled from rabbits varying in age and sex (82 kits, 6 does, 6 bucks, 19 adult rabbits), which were aged 2 wk (14 suckling rabbits), 6 and 8 wk (68 weaning and post-weaning rabbits), adult rabbits (31, one year). Faecal mixtures were sampled into sterile packs with faeces from 5-6 animals in each. The total count of enterococci from these Pannon White rabbits reached, on av. 5.28±0.29 colony forming units/g (log10). Among the 19 enterococci, 14 E. faecalis and 5 E. faecium were detected using 3 identification methods. The enterococci were mostly resistant to antibiotics. They were non-haemolytic, Dnase and urease negative. They did not form biofilm. They were free of the hylEfm gene and IS16 genes; the most frequently detected genes were gelE, efaAfm, efaAfs. Based on these results, E. faecium EF9a was selected for further analysis.

[EN] Properties of enterococci isolated from the Hungarian breed Pannon White were studied to spread knowledge regarding the properties of microbiota in rabbits from the basic research standpoint and to select a beneficial candidate for application in husbandry. Faeces from 113 Pannon White rabbits (mostly maternal line and some paternal line) were collected. They were sampled from rabbits varying in age and sex (82 kits, 6 does, 6 bucks, 19 adult rabbits), which were aged 2 wk (14 suckling rabbits), 6 and 8 wk (68 weaning and post-weaning rabbits), adult rabbits (31, one year). Faecal mixtures were sampled into sterile packs with faeces from 5-6 animals in each. The total count of enterococci from these Pannon White rabbits reached, on av. 5.28±0.29 colony forming units/g (log10). Among the 19 enterococci, 14 E. faecalis and 5 E. faecium were detected using 3 identification methods. The enterococci were mostly resistant to antibiotics. They were non-haemolytic, Dnase and urease negative. They did not form biofilm. They were free of the hylEfm gene and IS16 genes; the most frequently detected genes were gelE, efaAfm, efaAfs. Based on these results, E. faecium EF9a was selected for further analysis.

We describe clonal shifts in vanA Enterococcus faecium isolates from clinical samples obtained from patients in Denmark from 2015 to the first quarter (Q1) of 2019. During Q1 2019, the vancomycin-variable enterococci (VVE) ST1421-CT1134 vanA E. faecium became the most dominant vanA E. faecium clone and has spread to all five regions in Denmark. Among 174 E. faecium isolates with vanA, vanB or vanA/vanB genes in Q1 2019, 44% belonged to this type.

[EN] Properties of enterococci isolated from the Hungarian breed Pannon White were studied to spread knowledge regarding the properties of microbiota in rabbits from the basic research standpoint and to select a beneficial candidate for application in husbandry. Faeces from 113 Pannon White rabbits (mostly maternal line and some paternal line) were collected. They were sampled from rabbits varying in age and sex (82 kits, 6 does, 6 bucks, 19 adult rabbits), which were aged 2 wk (14 suckling rabbits), 6 and 8 wk (68 weaning and post-weaning rabbits), adult rabbits (31, one year). Faecal mixtures were sampled into sterile packs with faeces from 5-6 animals in each. The total count of enterococci from these Pannon White rabbits reached, on av. 5.28±0.29 colony forming units/g (log10). Among the 19 enterococci, 14 E. faecalis and 5 E. faecium were detected using 3 identification methods. The enterococci were mostly resistant to antibiotics. They were non-haemolytic, Dnase and urease negative. They did not form biofilm. They were free of the hylEfm gene and IS16 genes; the most frequently detected genes were gelE, efaAfm, efaAfs. Based on these results, E. faecium EF9a was selected for further analysis.

Preliminary laboratory work has shown that electrochemical oxidation (ECO) is a promising technology for disinfection of harvested stormwater. This paper focuses on understanding how stormwater chemistry (e.g. pH, chloride, bicarbonate, ammonia and total organic carbon - that can vary substantially between sites) impacts the disinfection performance of ECO. Real stormwater samples from four different urban catchments were collected and tested for ECO performance in disinfecting stormwater pathogens using a boron doped diamond anode under the current density of 4.2 mA/cm2. Results showed that total disinfection of indigenous Escherichia coli (E. coli), as well as three different stormwater pathogens (Enterococci, Campylobacter and C. perfringens) was achievable for all four tested stormwater within 30 min. Compared to the synthetic stormwater, lower disinfection rates were observed in real stormwater which has more complex chemistry. Stormwater chloride concentration was the only tested parameter that had significant impact on the treatment performance, with higher initial stormwater chloride concentration leading to an increased disinfection rate. Disinfection by-products in the treated stormwater were well below the Australian Drinking Water Guideline value for health, with its production level positively correlated to the pH values of stormwater.

When beach water monitoring programs identify poor water quality, the causes are frequently unknown. We hypothesize that management policies play an important role in the frequency of fecal indicator bacteria (FIB) exceedances (enterococci and fecal coliform) at recreational beaches. To test this hypothesis we implemented an innovative approach utilizing large amounts of monitoring data (n > 150,000 measurements per FIB) to determine associations between the frequency of contaminant exceedances and beach management practices. The large FIB database was augmented with results from a survey designed to assess management policies for 316 beaches throughout the state of Florida. The FIB and survey data were analyzed using t-tests, ANOVA, factor analysis, and linear regression. Results show that beach geomorphology (beach type) was highly associated with exceedance of regulatory standards. Low enterococci exceedances were associated with open coast beaches (n = 211) that have sparse human densities, no homeless populations, low densities of dogs and birds, bird management policies, low densities of seaweed, beach renourishment, charge access fees, employ lifeguards, without nearby marinas, and those that manage storm water. Factor analysis and a linear regression confirmed beach type as the predominant factor with secondary influences from grooming activities (including seaweed densities and beach renourishment) and beach access (including charging fees, employing lifeguards, and without nearby marinas). Our results were observable primarily because of the very large public FIB database available for analyses; similar approaches can be adopted at other beaches. The findings of this research have important policy implications because the selected beach management practices that were associated with low levels of FIB can be implemented in other parts of the US and around the world to improve recreational beach water quality., Green Open Access added to TU Delft Institutional Repository ‘You share, we take care!’ – Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public., Environmental Fluid Mechanics

Although many U.S. homes rely on private wells, few studies have investigated the quality of these water sources. This cross-sectional study evaluated private well water quality in Maryland, and explored possible environmental sources that could impact water quality. Well water samples (n = 118) were collected in four Maryland counties and were analyzed for microbiological and chemical contaminants. Data from the U.S. Census of Agriculture were used to evaluate associations between the presence of animal feeding operations and well water quality at the zip code level using logistic regression. Overall, 43.2% of tested wells did not meet at least one federal health-based drinking water standard. Total coliforms, fecal coliforms, enterococci, and Escherichia coli were detected in 25.4%, 15.3%, 5.1%, and 3.4% of tested wells, respectively. Approximately 26%, 3.4%, and <1% of wells did not meet standards for pH, nitrate-N, and total dissolved solids, respectively. There were no statistically significant associations between the presence of cattle, dairy, broiler, turkey, or aquaculture operations and the detection of fecal indicator bacteria in tested wells. In conclusion, nearly half of tested wells did not meet federal health-based drinking water standards, and additional research is needed to evaluate factors that impact well water quality. However, homeowner education on well water testing and well maintenance could be important for public health.

Pathogenic ability has been extensively studied in clinical enterococci, but to a lesser extent in community-derived ones. Most studies to date in enterococci from healthy infants have been focused on Enterococcus faecalis, despite the growing concern about nosocomial infections caused by E. faecium. In this work, we studied the antibiotic resistance and virulence determinants of 26 E. faecalis and 15 E. faecium intestinal isolates from Spanish healthy breastfed infants. Overall, commensal enterococci studied contained antibiotic resistance and virulence genes, although their patterns were not according to those described for antibiotic-resistant hospital-associated enterococci. None of the isolates was resistant to vancomycin, although the majority showed resistance to some antibiotics. E. faecalis isolates harbored considerably more virulence determinants than E. faecium isolates, but some genes linked to colonization were abundant in both species. Hemolysin activity was not detected in any of the isolates; and the gelatinase gene, when present, was silent in E. faecium, whereas gelatinase activity occurred in half of the E. faecalis isolates studied. These results suggest an ambivalent role of some virulence determinants as elements of pathogenesis.

When beach water monitoring programs identify poor water quality, the causes are frequently unknown. We hypothesize that management policies play an important role in the frequency of fecal indicator bacteria (FIB) exceedances (enterococci and fecal coliform) at recreational beaches. To test this hypothesis we implemented an innovative approach utilizing large amounts of monitoring data (n > 150,000 measurements per FIB) to determine associations between the frequency of contaminant exceedances and beach management practices. The large FIB database was augmented with results from a survey designed to assess management policies for 316 beaches throughout the state of Florida. The FIB and survey data were analyzed using t-tests, ANOVA, factor analysis, and linear regression. Results show that beach geomorphology (beach type) was highly associated with exceedance of regulatory standards. Low enterococci exceedances were associated with open coast beaches (n = 211) that have sparse human densities, no homeless populations, low densities of dogs and birds, bird management policies, low densities of seaweed, beach renourishment, charge access fees, employ lifeguards, without nearby marinas, and those that manage storm water. Factor analysis and a linear regression confirmed beach type as the predominant factor with secondary influences from grooming activities (including seaweed densities and beach renourishment) and beach access (including charging fees, employing lifeguards, and without nearby marinas). Our results were observable primarily because of the very large public FIB database available for analyses; similar approaches can be adopted at other beaches. The findings of this research have important policy implications because the selected beach management practices that were associated with low levels of FIB can be implemented in other parts of the US and around the world to improve recreational beach water quality., Green Open Access added to TU Delft Institutional Repository ‘You share, we take care!’ – Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public., Environmental Fluid Mechanics

The objective of this paper was to emphasise how faecal bacteria persist in canal sediment and relate abiotic factors in a pen-urban canal. Mann-Whitney tests confirmed that there is a considerable difference in water quality of upstream and downstream ends of the canal, which supports the claim that water quality is affected by wastewater discharge. Strong positive correlations between enterococci and organic matter, TOC and all nitrogen forms in sediment show that the large amount of organic matter in sediment is linked to high enterococcal counts. High counts of E. coli in sediment, as well as weak correlations between E. coli and sediment quality parameters, together with water quality changes, indicate a 'fresh' pollution of aquatic system. In conclusion, our study suggests that wastewater discharge makes a long-term contribution to the increased content of E. cob and enterococci in the Palic-Ludas canal.

To improve microbial water quality and to prevent waterborne disease outbreaks, knowledge on the fate and transport of contaminants and on the contributions from different faecal sources to the total contamination is essential. The fate and transport of faecal indicators E. coli and enterococci within the Betna River in Bangladesh were simulated using a coupled hydrodynamic and water quality model. The hydrodynamic model for the river was set up, calibrated and validated with water level and discharge in our earlier study. In this study, the hydrodynamic model was further validated using measured water temperature and salinity and coupled with the water quality module. Bacterial load data from various faecal sources were collected and used as input in the water quality model. The model output corresponded very well with the measured E. coli and enterococci concentrations in the river; the Root Mean Square Error and the Nash-Sutcliffe efficiency for Log10-transformed concentrations were found to be 0.23 (Log10 CFU/100 ml) and 0.84 for E. coli, and 0.19 (Log10 CFU/100 ml) and 0.86 for enterococci, respectively. Then, the sensitivity of the model was tested by removing one process or forcing at a time. These simulations revealed that the microbial decay, the upstream concentrations and the discharge of untreated wastewater were the primary factors controlling the concentrations in the river, while wind and the contribution from the diffuse sources (i.e. urban and agricultural runoff) were unlikely to have a major influence. Finally, the model was applied to investigate the influence of wastewater treatment on the bacteria concentrations. This revealed that wastewater treatment would result in a considerable improvement of the microbial water quality of the Betna River. This paper demonstrates the application of a comprehensive state-of-art model in a river in a data-poor tropical area. The model can potentially be applied to other watersheds and can help in formulating

The present study aimed at identifying and assessing antimicrobial resistance of Enterococcus spp. isolated from small and medium enterprise slaughterhouses in Kenya. In total, 67 isolates were recovered from 48 of 195 samples examined from beef carcasses, personnel, and cutting equipment in five small and medium enterprise slaughterhouses. The isolates were identified by using matrix-assisted laser desorption-ionization time of flight mass spectrometry and screened thereafter for their resistance against 12 antibiotics by using a disk diffusion assay. The isolates (n=67) included Enterococcus faecalis (41.8%), Enterococcus mundtii (17.9%), Enterococcus thailandicus (13.4%), Enterococcus faechun (9.0%), Enterococcus hirae (7.5%), Enterococcus casseliflavus (6.0%), and Enterococcus devriesei (4.5%). None of the isolates were resistant to ciprofloxacin, penicillin, ampicillin, vancomycin, nitrofunuraoin, teicoplanin, linezohd, and levofioxacin. Resistance to rifampin (46.3%), erythromycin (23.9%), tetracycline (20.9%), and chloramphenicol (7.5%) was distributed among six of the seven species. All E. thadandicus were resistant to rifampin, erythromycin, and tetracycline. E. faecalis was resistant to rifampin (60.7%), tetracycline (17.9%), erythromycin (14.3%), and chloramphenicol (10.7%). Resistance to two or three antibiotics was observed in 26.9% of the enterococci isolates. The isolation of enterococci that are resistant to clinically relevant antibiotics, such as erythromycin, is of a serious concern given the role enterococci play in the transfer of antibiotic resistance genes.

© The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Marine Pollution Bulletin 121 (2017): 160-167, doi:10.1016/j.marpolbul.2017.05.024., Large databases of fecal indicator bacteria (FIB) measurements are available for coastal waters. With the assistance of satellite imagery, we illustrated the power of assessing data for many sites by evaluating beach features such as geomorphology, distance from rivers and canals, presence of piers and causeways, and degree of urbanization coupled with the enterococci FIB database for the state of Florida. We found that beach geomorphology was the primary characteristic associated with enterococci levels that exceeded regulatory guidelines. Beaches in close proximity to marshes or within bays had higher enterococci exceedances in comparison to open coast beaches. For open coast beaches, greater enterococci exceedances were associated with nearby rivers and higher levels of urbanization. Piers and causeways had a minimal contribution, as their effect was often overwhelmed by beach geomorphology. Results can be used to understand the potential causes of elevated enterococci levels and to promote public health., The early portion of this work was funded in part by the NSF–NIEHS Oceans and Human Health Program (NIEHS #P50 ES12736 and NSF #OCE0432368/0911373/1127813).

© The Author(s), 2017. This is the author's version of the work. It is posted here under a nonexclusive, irrevocable, paid-up, worldwide license granted to WHOI. It is made available for personal use, not for redistribution. The definitive version was published in Marine Pollution Bulletin 121 (2017): 160-167, doi:10.1016/j.marpolbul.2017.05.024., Large databases of fecal indicator bacteria (FIB) measurements are available for coastal waters. With the assistance of satellite imagery, we illustrated the power of assessing data for many sites by evaluating beach features such as geomorphology, distance from rivers and canals, presence of piers and causeways, and degree of urbanization coupled with the enterococci FIB database for the state of Florida. We found that beach geomorphology was the primary characteristic associated with enterococci levels that exceeded regulatory guidelines. Beaches in close proximity to marshes or within bays had higher enterococci exceedances in comparison to open coast beaches. For open coast beaches, greater enterococci exceedances were associated with nearby rivers and higher levels of urbanization. Piers and causeways had a minimal contribution, as their effect was often overwhelmed by beach geomorphology. Results can be used to understand the potential causes of elevated enterococci levels and to promote public health., The early portion of this work was funded in part by the NSF–NIEHS Oceans and Human Health Program (NIEHS #P50 ES12736 and NSF #OCE0432368/0911373/1127813).

Enterococci are widespread bacteria forming the third largest genus among lactic acid bacteria. Some possess probiotic properties or they can produce beneficial proteinaceous antimicrobial substances called enterocins. On the other hand, some enterococci produce biogenic amines (BAs), so this study is focused on the sensitivity to enterocins of biogenic amine-producing faecal enterococci from ostriches and pheasants. Altogether, 60 enterococci isolated from faeces of ostriches and pheasants were tested for production of BAs. This target of the identified enterococci involved 46 strains selected from 140 ostriches and 17 from 60 pheasants involving the species Enterococcus hirae, E. faecium, E. faecalis, and E. mundtii. Although BAs histamine, cadaverine, putrescine, and tryptamine were not detected in the enterococci tested, in general high BA production by the tested enterococci was noted. The species E. hirae formed the majority of the enterococcal strains from ostrichs faeces (34 strains). High production of tyramine (TYM) was measured with an average amount of 958.16 ± 28.18 mg/ml. Among the enterococci from pheasants, the highest was production of TYM compared to phenylethylamine, spermidine, and spermine. Enterococci featured high BA production; however, they were sensitive to seven enterocins with inhibition activity ranging from 100 up to 25,600 AU/ml. © 2017, Springer Science+Business Media New York.

Enterococciwere detected occasionally in 100 L samples ofwater abstracted froma shallowaquifer in a natural dune infiltration area for drinking water production. Enterococcus moraviensis was the species most frequently identified in these samples. Because there are no existing reports of faecal sources of E. moraviensis and the closely related E. hemoperoxidus, this study aimed to find such sources of these two species in the dunes. Faecal samples from various animal species living in the vicinity of abstraction wells, were analysed for enterococci on Slanetz and Bartley Agar. From these samples, enterococci isolates (1,386 in total) were subsequently identified using matrix assisted laser desorption ionizationtimeof flight (MALDI-TOF)mass spectrometry. E.moraviensiswas found in the faeces of geese, foxes and rabbits. Also, E. haemoperoxidus was isolated from goose faeces. Using hierarchical clustering, the species composition of Enterococcus spp. isolated from abstracted water formed one cluster with the species composition found in geese droppings. A sanitary survey supported the indication that feral geese may provide a substantial faecal load in particular parts of this dune infiltration area, close to the water abstraction system. This study confirms the faecal origin of E.moraviensis and E. haemoperoxidus from specific animals, which strengthens their significance as faecal indicators., Sanitary Engineering

In the production of fermented feed, each crop can be contaminated with a variety of microorganisms that may produce natural pollutants. Biogenic amines, mycotoxins, and undesirable organic acids can decrease health feed safety. The aim of this study was to compare the counts of microorganisms, levels of biogenic amines, and the mycotoxins in forage legumes, and also to compare the occurrence of microorganisms and levels of mycotoxins in green fodder and subsequently produced silage and the influence of additives on the content of natural harmful substances in silage. The experimental plot was located in Troubsko and Vatín, in the Czech Republic. Two varieties of Medicago sativa and one variety of Trifolium pratense were compared. Green fodder and subsequently produced silage reaching up to 23% of dry matter were evaluated and prepared using a bio-enzymatic additive and a chemical additive. Green fodder of Medicago sativa was more contaminated by Enterococci than Trifolium pratense fodder. The obvious difference was determined by the quality of silage leachate. The silage prepared from Medicago sativa fodder was more contaminated with butyric acid. Fungi were present in higher counts in the anaerobic environment of green fodder and contaminated it with zearalenone and deoxynivalenol. Lower counts of fungi were found in silage, although the zearalenone content did not change. Lower content of deoxynivalenol was detected in silage, compared with green fodder. Silages treated with a chemical additive were found not to contain butyric acid. Lower ethanol content was determined, and the tendency to reduce the risk of biogenic amines occurrence was evident. The additives proved to have no influence on the content of mycotoxins.

In the production of fermented feed, each crop can be contaminated with a variety of microorganisms that may produce natural pollutants. Biogenic amines, mycotoxins, and undesirable organic acids can decrease health feed safety. The aim of this study was to compare the counts of microorganisms, levels of biogenic amines, and the mycotoxins in forage legumes, and also to compare the occurrence of microorganisms and levels of mycotoxins in green fodder and subsequently produced silage and the influence of additives on the content of natural harmful substances in silage. The experimental plot was located in Troubsko and Vatín, in the Czech Republic. Two varieties of Medicago sativa and one variety of Trifolium pratense were compared. Green fodder and subsequently produced silage reaching up to 23% of dry matter were evaluated and prepared using a bio-enzymatic additive and a chemical additive. Green fodder of Medicago sativa was more contaminated by Enterococci than Trifolium pratense fodder. The obvious difference was determined by the quality of silage leachate. The silage prepared from Medicago sativa fodder was more contaminated with butyric acid. Fungi were present in higher counts in the anaerobic environment of green fodder and contaminated it with zearalenone and deoxynivalenol. Lower counts of fungi were found in silage, although the zearalenone content did not change. Lower content of deoxynivalenol was detected in silage, compared with green fodder. Silages treated with a chemical additive were found not to contain butyric acid. Lower ethanol content was determined, and the tendency to reduce the risk of biogenic amines occurrence was evident. The additives proved to have no influence on the content of mycotoxins.

In the production of fermented feed, each crop can be contaminated with a variety of microorganisms that may produce natural pollutants. Biogenic amines, mycotoxins, and undesirable organic acids can decrease health feed safety. The aim of this study was to compare the counts of microorganisms, levels of biogenic amines, and the mycotoxins in forage legumes, and also to compare the occurrence of microorganisms and levels of mycotoxins in green fodder and subsequently produced silage and the influence of additives on the content of natural harmful substances in silage. The experimental plot was located in Troubsko and Vatín, in the Czech Republic. Two varieties of Medicago sativa and one variety of Trifolium pratense were compared. Green fodder and subsequently produced silage reaching up to 23% of dry matter were evaluated and prepared using a bio-enzymatic additive and a chemical additive. Green fodder of Medicago sativa was more contaminated by Enterococci than Trifolium pratense fodder. The obvious difference was determined by the quality of silage leachate. The silage prepared from Medicago sativa fodder was more contaminated with butyric acid. Fungi were present in higher counts in the anaerobic environment of green fodder and contaminated it with zearalenone and deoxynivalenol. Lower counts of fungi were found in silage, although the zearalenone content did not change. Lower content of deoxynivalenol was detected in silage, compared with green fodder. Silages treated with a chemical additive were found not to contain butyric acid. Lower ethanol content was determined, and the tendency to reduce the risk of biogenic amines occurrence was evident. The additives proved to have no influence on the content of mycotoxins.

In the production of fermented feed, each crop can be contaminated with a variety of microorganisms that may produce natural pollutants. Biogenic amines, mycotoxins, and undesirable organic acids can decrease health feed safety. The aim of this study was to compare the counts of microorganisms, levels of biogenic amines, and the mycotoxins in forage legumes, and also to compare the occurrence of microorganisms and levels of mycotoxins in green fodder and subsequently produced silage and the influence of additives on the content of natural harmful substances in silage. The experimental plot was located in Troubsko and Vatín, in the Czech Republic. Two varieties of Medicago sativa and one variety of Trifolium pratense were compared. Green fodder and subsequently produced silage reaching up to 23% of dry matter were evaluated and prepared using a bio-enzymatic additive and a chemical additive. Green fodder of Medicago sativa was more contaminated by Enterococci than Trifolium pratense fodder. The obvious difference was determined by the quality of silage leachate. The silage prepared from Medicago sativa fodder was more contaminated with butyric acid. Fungi were present in higher counts in the anaerobic environment of green fodder and contaminated it with zearalenone and deoxynivalenol. Lower counts of fungi were found in silage, although the zearalenone content did not change. Lower content of deoxynivalenol was detected in silage, compared with green fodder. Silages treated with a chemical additive were found not to contain butyric acid. Lower ethanol content was determined, and the tendency to reduce the risk of biogenic amines occurrence was evident. The additives proved to have no influence on the content of mycotoxins.

In the production of fermented feed, each crop can be contaminated with a variety of microorganisms that may produce natural pollutants. Biogenic amines, mycotoxins, and undesirable organic acids can decrease health feed safety. The aim of this study was to compare the counts of microorganisms, levels of biogenic amines, and the mycotoxins in forage legumes, and also to compare the occurrence of microorganisms and levels of mycotoxins in green fodder and subsequently produced silage and the influence of additives on the content of natural harmful substances in silage. The experimental plot was located in Troubsko and Vatín, in the Czech Republic. Two varieties of Medicago sativa and one variety of Trifolium pratense were compared. Green fodder and subsequently produced silage reaching up to 23% of dry matter were evaluated and prepared using a bio-enzymatic additive and a chemical additive. Green fodder of Medicago sativa was more contaminated by Enterococci than Trifolium pratense fodder. The obvious difference was determined by the quality of silage leachate. The silage prepared from Medicago sativa fodder was more contaminated with butyric acid. Fungi were present in higher counts in the anaerobic environment of green fodder and contaminated it with zearalenone and deoxynivalenol. Lower counts of fungi were found in silage, although the zearalenone content did not change. Lower content of deoxynivalenol was detected in silage, compared with green fodder. Silages treated with a chemical additive were found not to contain butyric acid. Lower ethanol content was determined, and the tendency to reduce the risk of biogenic amines occurrence was evident. The additives proved to have no influence on the content of mycotoxins.

The understanding of mechanisms of interactions between various bacterial cell surface proteins and host receptors has become imperative for the study of the health promoting features of probiotic enterococci. This study, for the first time, describes a novel enterococcal aggregation protein, AggE, from Enterococcus faeciurn BGGO9-28, selected from a laboratory collection of enterococcal isolates with auto aggregation phenotypes. Among them, En. faecium BGG09-28 showed the strongest auto -aggregation, adhesion to components of ECM and biofilm formation. Novel aggregation promoting factor AggE, a protein of 178.1 kDa, belongs to the collagen -binding superfamily of proteins and shares similar architecture with previously discovered aggregation factors from lactic acid bacteria (LAB). Its expression in heterologous enterococcal and lactococcal hosts demonstrates that the aggE gene is sufficient for cell aggregation. The derivatives carrying aggE exhibited the ten times higher adhesion ability to collagen and fibronectin, possess about two times higher adhesion to mucin and contribute to the increase of biofilm formation, comparing to the control strains. Analysis for the presence of virulence factors (cytolysin and gelatinase production), antibiotic resistance (antibiotic susceptibility) and genes (cylA, egg, gelE, esp, hyiN, ace, efaks, and efagn) showed that BGG09-28 was sensitive to all tested antibiotics, without hemolytic or gelatinase activity. This strain does not carry any of the tested genes encoding for known virulence factors. Results showed that BGGO9-28 was resistant to low pH and high concentrations of bile salts. Also, it adhered strongly to the Caco-2 human epithelial cell line. In conclusion, the results of this study indicate that the presence of AggE protein on the cell surface in enterococci is a desirable probiotic feature.