Energy and protein levels in dairy cow diets to recover milk ethanol stability.

The objective of this study was to evaluate the effect of energy and protein in the diet on the recovery of milk ethanol stability (MES) induced by feed restriction. Twelve Holstein and Holstein x Jersey crossbred cows with an average of 146 ± 50 d in milk, 575.4 ± 70 kg of body weight, and 18.93 ± 5.46 kg/d of milk yield were distributed in a 3x3 Latin square design with 3 treatments and 3 experimental periods. Each experimental period lasted 24 d, comprising 3 phases: a 13-d adaptation phase (100E+100P), a 4-d induction phase for milk ethanol instability (50E+50P), and a 7-d recovery phase for MES (3 treatments). The 3 treatments during the recovery phase consisted of 3 diets aiming to meet the requirements of energy and protein (100E+100P), only energy (100E+50P), or only protein (50E+100P). The diet during the adaptation and induction phases was common for all cows. The energy and protein levels to meet each cow's requirements were based on the group average. Restriction of energy and protein reduced dry matter, crude protein, and total digestive nutrient intake for cows fed 100E+50P and 50E+100P. The lowest body weight was observed for cows fed 50E+100P, with no difference for body condition score. During the induction phase, MES "was" reduced by 9 percentage units. Cows fed 100E+100P recovered MES in the first days of the recovery phase, while 100E+50P slightly improved MES, and 50E+100P had a constant decrease in MES. Cows fed 100E+50P and 50E+100P produced, respectively, 3.6 and 5.9 kg less milk than those fed 100E+100P. The 50E+100P treatment exhibited the highest milk fat content and somatic cell score, along with the lowest milk lactose content. Protein content was higher in the 100E+100P treatment. Cows fed 50E+100P showed higher serum albumin levels compared with those on the 100E+100P treatment, not differing from the 100E+50P treatment. We concluded that the complete recovery of MES in cows with feed restrictions is possible only by supplying both the energy and protein requirements in the cows' diet. However, restricting energy intake poses a greater limitation on MES recovery compared with restricting protein.

The impact of potassium peroxymonosulphate and chlorinated cyanurates on biofilms of Stenotrophomonas maltophilia: effects on biofilm control, regrowth, and mechanical properties.

The activity of two chlorinated isocyanurates (NaDCC and TCCA) and peroxymonosulphate (OXONE) was evaluated against biofilms of Stenotrophomonas maltophilia, an emerging pathogen isolated from drinking water (DW), and for the prevention of biofilm regrowth. After disinfection of pre-formed 48 h-old biofilms, the culturability was reduced up to 7 log, with OXONE, TCCA, and NaDCC showing more efficiency than free chlorine against biofilms formed on stainless steel. The regrowth of biofilms previously exposed to OXONE was reduced by 5 and 4 log CFU cm-2 in comparison to the unexposed biofilms and biofilms exposed to free chlorine, respectively. Rheometry analysis showed that biofilms presented properties of viscoelastic solid materials, regardless of the treatment. OXONE reduced the cohesiveness of the biofilm, given the significant decrease in the complex shear modulus (G*). AFM analysis revealed that biofilms had a fractured appearance and smaller bacterial aggregates dispersed throughout the surface after OXONE exposure than the control sample. In general, OXONE has been demonstrated to be a promising disinfectant to control DW biofilms, with a higher activity than chlorine. The results also show the impact of the biofilm mechanical properties on the efficacy of the disinfectants in biofilm control.

Biofilm formation under high shear stress increases resilience to chemical and mechanical challenges.

The effect that the hydrodynamic conditions under which biofilms are formed has on their persistence is still unknown. This study assessed the behaviour of Pseudomonas fluorescens biofilms, formed on stainless steel under different shear stress (τw) conditions (1, 2 and 4 Pa), to chemical (benzalkonium chloride - BAC, glutaraldehyde - GLUT and sodium hypochlorite - SHC) and mechanical (20 Pa) treatments (alone and combined). The biofilms formed under different τw  showed different structural characteristics. Those formed under a higher τw were invariably more tolerant to chemical and mechanical stresses. SHC was the biocide which caused the highest biofilm killing and removal, followed by BAC. The sequential exposure to biocides and mechanical stress was found to be insufficient for effective biofilm control. A basal layer containing biofilm cells mostly in a viable state remained on the surface of the cylinders, particularly for the 2 and 4 Pa-generated biofilms.

Photodynamic therapy and combinatory treatments for the control of biofilm-associated infections.

The advent of antimicrobial resistance has added considerable impact to infectious diseases both in the number of infections and healthcare costs. Furthermore, the relentless emergence of multidrug-resistant bacteria, particularly in the biofilm state, has made mandatory the discovery of new alternative antimicrobial therapies that are capable to eradicate resistant bacteria and impair the development of new forms of resistance. Amongst the therapeutic strategies for treating biofilms, antimicrobial photodynamic therapy (aPDT) has shown great potential in inactivating several clinically relevant micro-organisms, including antibiotic-resistant 'priority bacteria' declared by the WHO as critical pathogens. Its antimicrobial effect is centred on the basis that harmless low-intensity light stimulates a non-toxic dye named photosensitizer, triggering the production of reactive oxygen species upon photostimulation. In addition, combination therapies of aPDT with other antimicrobial agents (e.g. antibiotics) have also drawn considerable attention, as it is a multi-target strategy. Therefore, the present review highlights the recent advances of aPDT against biofilms, also covering progress on combination therapy.

Influence of surface copper content on Stenotrophomonas maltophilia biofilm control using chlorine and mechanical stress.

This work aimed to evaluate the action of materials with different copper content (0, 57, 96 and 100%) on biofilm formation and control by chlorination and mechanical stress. Stenotrophomonas maltophilia isolated from drinking water was used as a model microorganism and biofilms were developed in a rotating cylinder reactor using realism-based shear stress conditions. Biofilms were characterized phenotypically and exposed to three control strategies: 10 mg l-1 of free chlorine for 10 min, an increased shear stress (a fluid velocity of 1.5 m s-1 for 30s), and a combination of both treatments. These shock treatments were not effective in biofilm control. The benefits from the use of copper surfaces was found essentially in reducing the numbers of non-damaged cells. Copper materials demonstrated better performance in biofilm prevention than chlorine. In general, copper alloys may have a positive public health impact by reducing the number of non-damaged cells in the water delivered after chlorine exposure.

The role of interleukins in vitiligo: a systematic review.

Vitiligo is a disorder of the skin that causes depigmentation and asymptomatic macules whose exact cause is still unclear. Although its aetiology is not fully elucidated, the main theory of its pathomechanism is that it is associated with the autoimmune process. There is few summarized information about the role of inflammatory mediators, as interleukins, in vitiligo, so our aim was to present a systematic review of the role of interleukins in vitiligo, focusing on interleukins. In this review, we included all studies assessing interleukin levels in vitiligo patients conducted up to June 2017. Quality assessment of these studies was performed using the Newcastle-Ottawa Scale (NOS). The interleukins mainly involved were IL-2, IL-4, IL-6, IL-10 and IL-17. The studies highlight the crucial role of IL-17 in the onset and progression of the disease, and its synergistic action with IL-2, IL-6 and IL-33. Dysregulated levels of the interleukins were also correlated with the stage of disease, the affected skin surface area, and indicated as the main factor for lymphocyte infiltration found in depigmented regions. These findings illustrate the growing need for new therapies targeting vitiligo and further research into the role of interleukins as an area of particular interest.

Klinefelter syndrome and fertility-Impact of X-chromosomal inheritance on spermatogenesis.

With the use of testicular sperm extraction (TESE), spermatozoa can be retrieved in about 30%-50% of men with Klinefelter syndrome (KS). The reason for the absence or presence of spermatozoa in half of the men with KS remains unknown. Therefore, the search for an objective marker for a positive prediction in finding spermatozoa is of significant clinical value to avoid unnecessary testicular biopsies in males with (mostly) low testicular volume and impaired testosterone. The objective of this study was to determine whether paternal or maternal inheritance of the additional X-chromosome can predict the absence or presence of spermatogenesis in men with KS. Men with KS who have had a testicular biopsy for diagnostic fertility workup TESE were eligible for inclusion. Buccal swabs from nine KS patients and parents (trios) were taken to compare X-chromosomal inheritance to determine the parental origin of both X-chromosomes in the males with KS. Spermatozoa were found in TESE biopsies 8 of 35 (23%) patients after performing a unilateral or bilateral TESE. Different levels of spermatogenesis (from the only presence of spermatogonia, up to maturation arrest or hypospermatogenesis) appeared to be present in 19 of 35 (54%) men, meaning that the presence of spermatogenesis not always yields mature spermatozoa. From the nine KS-trios that were genetically analysed for X-chromosomal inheritance origin, no evidence of a correlation between the maternal or paternal origin of the additional X-chromosome and the presence of spermatogenesis was found. In conclusion, the maternal or paternal origin of the additional X-chromosome in men with KS does not predict the presence or absence of spermatogenesis.

Effect of Zirconia Pigmentation on Translucency.

PURPOSE: the aim of this study was to investigate the effect of shading on translucency and mean grain size of three Zirconia systems (Lava Frame, Ice Zirkon Translucent and Prettau Zirkon). MATERIALS AND METHODS: specimens pigmented in A1, A4 and a non-pigmented specimen was evaluated with Scanning Electron Microscopy and spectrophotometery. RESULTS: the transmittance ranged between 0.18 and 0.52, in Ice Zirkon Translucent A4 and Prettau Zircon non-pigmented, respectively. The pigmentation A1 and A4 significantly reduced the light transmission in all the systems evaluated (p⟨0.05), with the exception of Lava A1. The light transmittance of each ceramic system is different from each other in the three pigmentations, except for the Prettau Zirkon A1, which did not show a significant difference compared with Lava A1 (p⟩0.05). The mean grain size increased with pigmentation. The mean grain size ranged between 0.43 µm, for Prettau Zirkon non-pigmented, and 0.82 µm for Prettau Zirkon A4. CONCLUSION: Light transmission and mean grain size was affected by pigmentation of dental zirconia ceramics.

Ordered La0.7Sr0.3MnO3 nanohole arrays fabricated on a nanoporous alumina template by pulsed laser ablation.

Highly ordered nanohole arrays of [Formula: see text] manganite have been synthesized using pulsed laser deposition on nanoporous alumina template. Their structure and phase formation were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), energy dispersive x-ray spectroscopy (EDX) and x-ray diffraction (XRD). The magnetic measurements were performed with respect to temperature and field and exhibit a ferromagnetic to paramagnetic transition at 284 K. In addition, the temperature dependence of electrical resistance was measured at different magnetic fields and an insulating phase throughout all the temperatures was observed. The low temperature ferromagnetic insulating state is discussed by the presence of a canted ferromagnetic state induced by the nanoholes. The present work shows the feasibility of combining both the nanoporous alumina template and pulsed laser ablation for the fabrication of perovskite manganite nanohole arrays which can also be extended to fabricate other multicomponent oxide nanohole materials.

Phytochemical profiling as a solution to palliate disinfectant limitations.

The indiscriminate use of biocides for general disinfection has contributed to the increased incidence of antimicrobial tolerant microorganisms. This study aims to assess the potential of seven phytochemicals (tyrosol, caffeic acid, ferulic acid, cinnamaldehyde, coumaric acid, cinnamic acid and eugenol) in the control of planktonic and sessile cells of Staphylococcus aureus and Escherichia coli. Cinnamaldehyde and eugenol showed antimicrobial properties, minimum inhibitory concentrations of 3-5 and 5-12 mM and minimum bactericidal concentrations of 10-12 and 10-14 mM against S. aureus and E. coli, respectively. Cinnamic acid was able to completely control adhered bacteria with effects comparable to peracetic acid and sodium hypochlorite and it was more effective than hydrogen peroxide (all at 10 mM). This phytochemical caused significant changes in bacterial membrane hydrophilicity. The observed effectiveness of phytochemicals makes them interesting alternatives and/or complementary products to commonly used biocidal products. Cinnamic acid is of particular interest for the control of sessile cells.

Magnetic liposomes based on nickel ferrite nanoparticles for biomedical applications.

Nickel ferrite nanoparticles with superparamagnetic behavior at room temperature were synthesized using a coprecipitation method. These magnetic nanoparticles were either covered with a lipid bilayer, forming dry magnetic liposomes (DMLs), or entrapped in liposomes, originating aqueous magnetoliposomes (AMLs). A new and promising method for the synthesis of DMLs is described. The presence of the lipid bilayer in DMLs was confirmed by FRET (Förster Resonance Energy Transfer) measurements between the fluorescent-labeled lipids NBD-C12-HPC (NBD acting as a donor) included in the second lipid layer and rhodamine B-DOPE (acceptor) in the first lipid layer. An average donor-acceptor distance of 3 nm was estimated. Assays of the non-specific interactions of magnetoliposomes with biological membranes (modeled using giant unilamellar vesicles, GUVs) were performed. Membrane fusion between both aqueous and dry magnetoliposomes and GUVs was confirmed by FRET, which is an important result regarding applications of these systems both as hyperthermia agents and antitumor drug nanocarriers.

Far-UV-C irradiation promotes synergistic bactericidal action against adhered cells of Escherichia coli and Staphylococcus epidermidis.

The contamination of indoor areas is a global health problem that can cause the dispersion of infectious diseases. In that sense, it is urgent to find new strategies applying a lower concentration of the traditional chemicals used for cleaning and disinfection. Ultraviolet radiation (UV), in particular far-UV-C (200-225 nm), has emerged as a successful, powerful, easy-to-apply, and inexpensive approach for bacterial eradication that still requires scientific assessment. This study investigated new strategies for disinfection based on far-UV-C (222 nm) combined with chlorine and mechanical cleaning, providing an innovative solution using low doses. The bactericidal activity of far-UV-C (222 nm) was tested at an intensity of irradiation from 78.4 µW/cm2 to 597.7 µW/cm2 (for 1 min) against Escherichia coli and Staphylococcus epidermidis adhered on polystyrene microtiter plates. It was further tested in combination with mechanical cleaning (ultrasounds for 1 min) and free chlorine (0.1, 0.5, and 1 mg/L for 5 min). The triple combination consisting of mechanical cleaning + free chlorine (0.5 mg/L) + far-UV-C (54 mJ/cm2) was tested against cells adhered to materials found in hospital settings and other public spaces: polyvinyl chloride (PVC), stainless steel (SS), and polyetheretherketone (PEEK). Disinfection with far-UV-C (54 mJ/cm2) and free chlorine at 0.5 mg/L for 5 min allowed a total reduction of culturable E. coli cells and a logarithmic reduction of 2.98 ± 0.03 for S. epidermidis. The triple combination of far-UV-C, free chlorine, and mechanical cleaning resulted in a total reduction of culturable cells for both adhered bacteria. Bacterial adhesion to PVC, SS, and PEEK occurred at distinct extents and influenced the bactericidal activity of the triple combination, with logarithmic reductions of up to three. The overall results highlight that, based on culturability assessment, far-UV-C (54 mJ/cm2) with chlorine (0.5 mg/L; 5 min) and mechanical cleaning (1 min) as an efficient disinfection strategy using mild conditions. The combination of culturability and viability assessment of disinfection is recommended to detect regrowth events and increase the effectiveness in microbial growth control.

Influence of the Adopted Balloon Modeling Strategies in the Stent Deployment Procedure: An In-Silico Analysis.

PURPOSE: As the promoter of the stent expansion, the balloon plays a very important role, offering a strong influence on the deployment process. Balloon-artery interaction is pointed as a probable cause of restenosis, stressing the relevance of balloon modeling when simulating the stenting procedure. In this work, an in-silico study of the balloon modeling strategies is performed. METHODS: Ultrasonic-microcasting is a novel technology that allows obtaining stents manufactured in magnesium alloys, being suggested as a promising solution. However, this technique demands superior stent strut thickness, which may have an impact on the stent deployment procedure. The influence of the balloon modeling is studied through the simulation of different balloon geometries (open- or taper-ended) and material constitutive model (linear elastic or hyperelastic) on the expanded configuration of a stent manufactured through ultrasonic-microcasting. RESULTS: The results obtained suggest that the choice of balloon type has small impact in terms of demanded pressure to inflate the balloon and in the stent final radius achieved at fully-expanded configuration. Additionally, it was proved that the balloon-type influences the stent expanded profile along its length and diameter as a result of the different deformation behavior exhibited by the balloon. CONCLUSION: The hyperelastic taper-ended balloon suggests being the model that better correlates with both experimental and clinical results regarding the expanded balloon profile during the procedure.

Health-related quality of life of inpatients and outpatients with TB in rural Malawi.

INTRODUCTION: Patients being treated for TB may suffer reductions in health-related quality of life (HRQoL). This study aims to assess the extent of such reductions and the trajectory of HRQoL over the course of treatment in rural Malawi.METHODS: We collected patient demographic and socioeconomic status, TB-related characteristics, and HRQoL data (i.e., EQ-5D and a visual analogue scale VAS) from adults (age ≥18 years) being treated for TB in 12 primary health centers and one hospital in rural Thyolo District, Malawi, from 2014 to 2016. Associations between HRQoL and patient characteristics were estimated using multivariable linear regression.RESULTS: Inpatients (n = 197) consistently showed lower median HRQoL scores and suffered more severe health impairments during hospitalization than outpatients (n = 156) (EQ5D and VAS: 0.79, 55 vs. 0.84, 70). Longer treatment duration was associated with higher HRQoL among outpatients (EQ5D: 0.034 increase per 2 months, 95%CI 0.012-0.057). We found no substantial associations between patients´ demographic and socioeconomic characteristics and HRQoL in this setting.CONCLUSION: HRQoL scores among patients receiving treatment for TB in rural Malawi differ by clinical setting and duration of treatment, with greater impairment among inpatients and those early in their treatment course.

Evolution of two-step magnetic transition on nanogranular Gd5Si1.3Ge2.7 thin film.

A multi-functional Gd5Si1.3Ge2.7 thin film deposited by pulsed laser ablation in the form of an ensemble of nanoparticles was studied for 18 thermal cycles via electron transport measurements together with structural and magnetic characterization. A general negative thermal dependency of the resistivity (ρ) is observed, which contrasts with the metallic-like behavior observed in bulk Gd5Si x Ge4-x compounds. This general trend is interrupted by a two-step, positive-slope transition in ρ(T) throughout the [150, 250] K interval, corresponding to two consecutive magnetic transitions: a fully coupled magnetostructural followed by a magnetic order on heating. An avalanche-like behavior is unveiled by the ∂ρ/∂T(T) curves and is explained based on the severe strains induced cyclically by the magnetostructural transition, leading to a cycling evolution of the transition onset temperature ([Formula: see text]/∂n ∼ 1.6 K/cycle, n being the number of cycles). Such behavior is equivalent to the action of a pressure of 0.56 kBar being formed and building up at every thermal cycle due to the large volume induced change across the magnetostructural transition. Moreover the thermal hysteresis, detected in both ρ and magnetization versus temperature curves, evolves significantly along the cycles, decreasing as n increases. This picture corroborates the thermal activation energy enhancement-estimated via an exponential fitting of the ∂ρ/∂T(T) in the avalanche regime. This work demonstrates the importance of using a short-range order technique, to probe both magnetic and magnetostructural transitions and their evolution with thermal cycles.

The scavenger receptor MARCO is involved in Leishmania major infection by CBA/J macrophages.

CBA/J mice are resistant to Leishmania major infection but are permissive to L. amazonensis infection. In addition, CBA/J macrophages control L. major but not L. amazonensis infection in vitro. Phagocytosis by macrophages is known to determine the outcome of Leishmania infection. Pattern recognition receptors (PRR) adorning antigen presenting cell surfaces are known to coordinate the link between innate and adaptive immunity. The macrophage receptor with collagenous structure (MARCO) is a PRR that is preferably expressed by macrophages and is capable of binding Gram-positive and Gram-negative bacteria. No research on the role of MARCO in Leishmania-macrophage interactions has been reported. Here, we demonstrate, for the first time, that MARCO expression by CBA/J macrophages is increased in response to both in vitro and in vivo L. major infections, but not to L. amazonensis infection. In addition, a specific anti-MARCO monoclonal antibody reduced L. major infection of macrophages by 30%-40% in vitro. The draining lymph nodes of anti-MARCO-treated mice displayed a reduced presence of immunolabelled parasite and parasite antigens, as well as a reduced inflammatory response. These results support the hypothesis that MARCO has a role in macrophage infection by L. major in vitro as well as in vivo.

Smoking habits in secondary school students.

BACKGROUND: Smoking is an important health risk in general, and responsible for diseases with significant mortality and morbidity. Smoking habits start early and adolescence is a notorious time for starting smoking. AIM AND METHODS: To assess knowledge on smoking and smoking habits in a population of adolescents in four Porto schools, using a confidential self administered questionnaire. Collected data were evaluated using the SPSS 1.2 statistics program (2004 version). RESULTS: A total of 1,770 students aged 11 - 21 (median 15.1 years), mainly female, (58%), answered. Most students (n=952, 54.6%) were unaware of signs or warnings against smoking in their schools. The great majority (n=1639, 92.7%) considered themselves well informed on the harmful effects of smoking, but only 6.7% could list three or more tobacco-associated health consequences, however. Parents and friends were seen as privileged sources of information. Among these students, 194 (11.1%) were smokers and the average started to smoke at the age of 15. The majority of these (n=111, 57.2%) had parents who smoked and 96.4% had friends who smoked, versus 83.1% of non-smokers, a statistically significant difference (p < 0.001). Pocket money was the means of acquiring cigarettes in 34.8%. Most (60.8%) considered themselves able to stop smoking at any time, while 11.4% of the smokers smoked more than one pack a day and 9.8% smoked the first cigarette within 5 minutes of waking, however. CONCLUSIONS: The percentage of smokers in this group of teenagers was considerable and indicators of nicotine dependence were found. Knowledge of the risks of smoking was poor and information on smoking given by schools had an apparently low and variable impact. Parents' and friends' behaviour may have a weighty impact on the decision to start smoking.

The role of surface copper content on biofilm formation by drinking water bacteria.

Copper pipes are conventionally used to supply tap water. Their role in biofilm prevention remains to be understood. This study evaluates the ability of selected surface materials with different copper contents (0, 57, 79, 87, 96, 100% of copper) to control biofilm formation and regrowth. Further experiments were performed to assess copper leaching and corrosion under conditions mimicking real plumbing systems. Acinetobacter calcoaceticus and Stenotrophomonas maltophilia isolated from a drinking water distribution system were used as model bacteria. All the copper materials showed positive results on the control of single and dual species biofilms presenting high reductions of bacterial culturability > 4 log CFU per cm2. The antimicrobial action of the selected materials seem not to be related to copper leaching or to the formation of reactive oxygen species. However, bacterial-copper contact demonstrated damage to bacterial membranes. The alloy containing 96% copper was the most promising surface in reducing biofilm culturability and viability, and was the only surface able to avoid the regrowth of single species biofilms when in contact with high nutrient concentrations. The alloy with 87% copper was shown to be unsuitable for use in chlorinated systems due to the high copper leaching observed when exposed to free chlorine. The presence of viable but non-culturable bacteria was remarkable, particularly in dual species biofilms. The overall results provide novel data on the role of copper alloys for use under chlorinated and unchlorinated conditions. Copper alloys demonstrated comparable or even higher biofilm control effects than elemental copper surfaces.

Prolonged exposure of Stenotrophomonas maltophilia biofilms to trace levels of clofibric acid alters antimicrobial tolerance and virulence.

The presence of pharmaceuticals in water sources, including in drinking water (DW), is increasingly being recognized as an emerging and global concern for the environment and public health. Based on the principles of the "One Health" initiative, the present work aims to understand the effects of clofibric acid (CA), a lipid regulator, on the behavior of a selected bacterium isolated from drinking water (DW). Biofilms of the opportunistic pathogen Stenotrophomonas maltophilia were exposed to CA for 12 weeks at 170 and 17000 ng/L. The effects of CA were evaluated on planktonic S. maltophilia susceptibility to chlorine and antibiotics (amoxicillin, ciprofloxacin, erythromycin, kanamycin, levofloxacin, oxacillin, spectinomycin, tetracycline and trimethoprim-sulfamethoxazole), biofilm formation, motility, siderophores production and on the adhesion and internalization of the human colon adenocarcinoma cell line (HT-29). It was found that CA did not affect planktonic S. maltophilia tolerance to chlorine exposure. Additionally, no effects were observed on biofilm formation, motility and siderophores production. However, biofilms formed after CA exposure were more tolerant to chlorine disinfection and lower CFU reductions were obtained. Of additional concern was the effect of CA exposure on S. maltophilia increased tolerance to erythromycin. CA exposure also slightly reduced S. maltophilia ability to invade HT-29 cells. In conclusion, this work reinforces the importance of studying the effects of non-antibiotic contaminants on the behavior of environmental microorganisms, particularly their role as drivers affecting resistance evolution and selection.