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Modern genetic approaches are powerful in providing access to diverse cell types in the brain and facilitating the study of their function. Here, we report a large set of driver and reporter transgenic mouse lines, including 23 new driver lines targeting a variety of cortical and subcortical cell populations and 26 new reporter lines expressing an array of molecular tools. In particular, we describe the TIGRE2.0 transgenic platform and introduce Cre-dependent reporter lines that enable optical physiology, optogenetics, and sparse labeling of genetically defined cell populations. TIGRE2.0 reporters broke the barrier in transgene expression level of single-copy targeted-insertion transgenesis in a wide range of neuronal types, along with additional advantage of a simplified breeding strategy compared to our first-generation TIGRE lines. These novel transgenic lines greatly expand the repertoire of high-precision genetic tools available to effectively identify, monitor, and manipulate distinct cell types in the mouse brain.
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Encéfalo/metabolismo , Técnicas de Inactivación de Genes/métodos , Genes Reporteros , Animales , Encéfalo/citología , Calcio/metabolismo , Línea Celular , Hibridación Fluorescente in Situ , Luz , Ratones , Ratones Transgénicos , Microscopía Fluorescente , Neuronas/metabolismo , Optogenética , ARN no Traducido/genética , Transgenes/genéticaRESUMEN
Interferons and central nervous system resident macrophages, microglia, are well-known for their respective roles in antiviral defense and phagocytosis. Using a classic experimental paradigm for examining activity-dependent neural plasticity, Escoubas, Dorman, et al. recently identified a role for microglial type I interferon signaling in the clearance of unwanted neurons during mouse brain development.
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Encéfalo , Interferón Tipo I , Microglía , Animales , Encéfalo/inmunología , Encéfalo/crecimiento & desarrollo , Interferón Tipo I/metabolismo , Interferón Tipo I/inmunología , Ratones , Microglía/inmunología , Microglía/metabolismo , Humanos , Transducción de Señal/inmunología , Neuronas/inmunología , Neuronas/metabolismo , Fagocitosis/inmunología , Plasticidad Neuronal/inmunologíaRESUMEN
Inflammation causes pain by shifting the balance of ionic currents in nociceptors toward depolarization, leading to hyperexcitability. The ensemble of ion channels within the plasma membrane is regulated by processes including biogenesis, transport, and degradation. Thus, alterations in ion channel trafficking may influence excitability. Sodium channel NaV1.7 and potassium channel KV7.2 promote and oppose excitability in nociceptors, respectively. We used live-cell imaging to investigate mechanisms by which inflammatory mediators (IM) modulate the abundance of these channels at axonal surfaces through transcription, vesicular loading, axonal transport, exocytosis, and endocytosis. Inflammatory mediators induced a NaV1.7-dependent increase in activity in distal axons. Further, inflammation increased the abundance of NaV1.7, but not of KV7.2, at axonal surfaces by selectively increasing channel loading into anterograde transport vesicles and insertion at the membrane, without affecting retrograde transport. These results uncover a cell biological mechanism for inflammatory pain and suggest NaV1.7 trafficking as a potential therapeutic target.
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Axones , Nociceptores , Ratas , Animales , Axones/metabolismo , Dolor/metabolismo , Inflamación/metabolismo , Mediadores de Inflamación/metabolismoRESUMEN
Precisely controlling the size of engineered biomolecules and pharmaceutical compounds is often critical to their function. Standard methods for size characterization, such as dynamic light scattering or size exclusion chromatography, can be sample intensive and may not provide the sensitivity needed for mass- or concentration-limited biological systems. Taylor dispersion analysis (TDA) is a proven analytical method for direct, calibration-free size determination which utilizes only nL-pL sample volumes. In TDA, diffusion coefficients, which are mathematically transformed to hydrodynamic radii, are determined by characterizing band broadening of an analyte under well-controlled laminar flow conditions. Here, we describe the design and development of a 3D printed instrument for TDA, which is the first such instrument to offer dual-point laser-induced fluorescence (LIF) detection. The instrument utilized a fully 3D printed eductor as a vacuum source for precise and stable pressure-driven flow within a capillary, evidenced by a linear response in generated static pressure to applied gas pressure (R2 = 0.997) and a 30-fold improvement in stability of static pressure (0.05% RSD) as compared to a standard mechanical pump (1.53%). Design aspects of the LIF detection system were optimized to maximize S/N for excitation and emission optical axes, and high sensitivity was achieved as evidenced by an 80 pM limit of detection for the protein R-Phycoerythrin and low nM limits of detection for three additional fluorophores. The utility of the instrument was demonstrated via sizing of R-Phycoerythrin at pM concentrations.
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Hidrodinámica , Ficoeritrina , Dispersión Dinámica de Luz , Rayos Láser , Impresión TridimensionalRESUMEN
The persistence of Phi6 (Φ6) bacteriophage on surfaces commonly encountered in consumer-facing environments was evaluated. Φ6 has been utilized as a surrogate for enveloped viruses, including SARS-CoV-2-the causative agent of COVID-19-due to structural similarities, biosafety level 1 (BSL-1) status, and ease of use. Φ6 persistence on fomites was evaluated by characterizing the impact of the inoculum matrix (artificial saliva, phosphate-buffered saline [PBS], tripartite), inoculum level (low and high), and surface type (nonporous-aluminum, stainless steel, plastic, touchscreen, vinyl; porous-wood). Φ6 was inoculated onto surfaces at low and high inoculum levels for each inoculum matrix and incubated (20.54 ± 0.48°C) for up to 168 h. Φ6 was eluted from the surface and quantified via the double agar overlay assay to determine virus survival over time. For nonporous surfaces inoculated with artificial saliva and PBS, significantly higher D values were observed with high inoculum application according to the 95% confidence intervals. In artificial saliva, D values ranged from 1.00 to 1.35 h at a low inoculum and 4.44 to 7.05 h at a high inoculum across inoculation matrices and surfaces. D values for Φ6, regardless of the inoculum level, were significantly higher in tripartite than in artificial saliva and PBS for nonporous surfaces. In contrast with artificial saliva or PBS, D values in tripartite at low inoculum (D values ranging from 45.8 to 72.8 h) were greater than those at high inoculum (D values ranging from 26.4 to 45.5 h) on nonporous surfaces. This study characterized the impact of the inoculum matrix, inoculum level, and surface type on Φ6 survival on various surfaces relevant to fomite transmission in public settings. IMPORTANCE An important consideration in virus contact transmission is the transfer rate between hands and surfaces, which is driven by several factors, including virus persistence on inanimate surfaces. This research characterized Φ6 persistence on surfaces commonly encountered in public settings based on various factors. The inoculum matrix, which simulates the route of transmission, can impact virus persistence, and three separate matrices were evaluated in this study to determine the impact on Φ6 persistence over time. The number of microorganisms has also been suggested to impact persistence, which was evaluated here to simulate real-world contamination scenarios on six surface types. Results from this study will guide future research utilizing Φ6 or other surrogates for enveloped viruses of public health concern.
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Bacteriófagos , COVID-19 , Virus , Fómites , Humanos , SARS-CoV-2 , Saliva ArtificialRESUMEN
AIMS: This study aimed to determine the extent of Phi6 (Φ6) transfer between skin and surfaces relevant to consumer-facing environments based on inoculum matrix, surface type and contact time. METHODS AND RESULTS: Φ6 transfer rates were determined from skin-to-fomite and fomite-to-skin influenced by inoculum matrix (artificial saliva and tripartite), surface type (aluminium, plastic, stainless steel, touchscreen, vinyl and wood) and contact time (5 and 10 s). Significant differences in estimated means were observed based on surface type (both transfer directions), inoculum matrix (skin-to-fomite) and contact time (both transfer directions). During a sequential transfer experiment from fomite-to-skin, the maximum number of consecutive transfer events observed was 3.33 ± 1.19, 2.33 ± 1.20 and 1.67 ± 1.21 for plastic, touchscreen and vinyl, respectively. CONCLUSIONS: Contact time significantly impacted Φ6 transfer rates, which may be attributed to skin absorption dynamics. Surface type should be considered for assessing Φ6 transfer rates. SIGNIFICANCE AND IMPACT OF THE STUDY: Although the persistence of Φ6 on fomites has been characterized, limited data are available regarding the transfer of Φ6 among skin and fomites. Determining Φ6 transfer rates for surfaces in consumer-facing environments based on these factors is needed to better inform future virus transmission mitigation strategies.
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Bacteriófagos , Humanos , Fómites , Acero Inoxidable , PlásticosRESUMEN
The microbial quality of agricultural water for fresh produce production is determined by the presence of the fecal indicator bacterium (FIB) Escherichia coli, despite poor correlations with pathogen presence. Additional FIB, such as enterococci, have been utilized for assessing water quality. The study objective was to determine the survival times (first time to detect zero or censored) of FIB (E. coli and enterococci), surrogates (Listeria innocua, Listeria seeligeri, Salmonella enterica serovar Typhimurium, and PRD1), and pathogens (four strains each of pathogenic E. coli and Listeria monocytogenes and five Salmonella serovars) simultaneously inoculated in freshwater mesocosms exposed to diel and seasonal variations. Six separate mesocosm experiments were conducted for ≤28 days each season, with samples (sediment/water) collected each day for the first 7 days and weekly thereafter. Microorganisms survived significantly longer in sediment than in water (hazard ratio [HR] for water/sediment is 2.2; 95% confidence interval [CI], 1.79 to 2.71). Also, FIB E. coli survived significantly longer than FIB enterococcus (HR for enterococci/E. coli is 12.9 [95% CI, 8.18 to 20.37]) after adjusting for the sediment/water and lake/river effects. Differences in the area under the curve (calculated from log CFU or PFU over time) were used to assess pathogen and surrogate survival in relation to FIB. Despite sample type (sediment/water) and seasonal influences, survival rates of pathogenic Salmonella serovars were similar to those of FIB E. coli, and survival rates of L. monocytogenes and pathogenic E. coli were similar to those of FIB enterococci. Further investigation of microbial survival in water and sediment is needed to determine which surrogates are best suited to assess pathogen survival in agricultural water used in irrigation water for fresh produce. IMPORTANCE Contamination of fresh produce via agricultural water is well established. This research demonstrates that survival of fecal indicator bacteria, pathogenic microorganisms, and other bacterial and viral surrogates in freshwater differs by sample type (sediment/water) and season. Our work highlights potential risks associated with pathogen accumulation and survival in sediment and the possibility for resuspension and contamination of agricultural water used in fresh produce production. Specifically, a greater microbial persistence in sediments than in water over time was observed, along with differences in survival among microorganisms in relation to the fecal indicator bacteria E. coli and enterococci. Previous studies compared data among microbial groups in different environments. Conversely, fecal indicator bacteria, surrogates, and pathogenic microorganisms were assessed within the same water and sediment mesocosms in the present study during four seasons, better representing the agricultural aquatic environment. These data should be considered when agricultural microbial water quality criteria in fresh produce operations are being determined.
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Riego Agrícola , Bacterias/aislamiento & purificación , Bacteriófago PRD1/aislamiento & purificación , Heces/microbiología , Agua Dulce/microbiología , Bacterias/virología , Microbiología del AguaRESUMEN
Epinephrine autoinjectors (EAIs) are important first aid medications for treating anaphylaxis. A 10-fold price increase over the past 12 years and evidence that expired EAIs may still contain significant doses of available epinephrine have motivated interest in the efficacy of expired EAIs as treatments of last resort. Degradation of expired EAIs, which can be caused by improper storage conditions, results in various degrees of discoloration of the epinephrine solution. Previous studies have determined that significant epinephrine remains available in expired EAIs, but these have only considered EAIs that show no discoloration. Here, we investigate the potential for colorimetric estimation of available epinephrine dose based on the degree of discoloration in expired EAIs. The correlation of available epinephrine dose and time since expiration date was poor (r = - 0.37), as determined by an industry standard UHPLC protocol. Visible absorbance of the samples integrated across the range 430-475 nm correlated well with available epinephrine dose (r = - 0.71). This wavelength corresponds to the blue channel of a typical smartphone camera Bayer filter. Smartphone camera images of the EAI solutions in various illumination conditions were analyzed to assign color indices representing the degree of discoloration. Color index of the samples showed similar correlation (|r| > 0.7) with available epinephrine dose as that of visible spectrophotometry. Smartphone imaging colorimetry is proposed as a potential point-of-use epinephrine dose estimator for expired and degraded EAIs. Graphical abstract.
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Anafilaxia/tratamiento farmacológico , Diagnóstico por Imagen/métodos , Estabilidad de Medicamentos , Almacenaje de Medicamentos/normas , Epinefrina/análisis , Autoadministración/métodos , Teléfono Inteligente/instrumentación , Epinefrina/administración & dosificación , Humanos , Inyecciones Intramusculares , Autoadministración/instrumentaciónRESUMEN
Microreactors have garnered widespread attention for their tunability and precise control of synthetic parameters to efficiently produce target species. Despite associated advances, a lack of online detection and optimization methods has stalled the progression of microfluidic reactors. Here we employ and characterize a total internal reflection transient absorption microscopy (TIRTAM) instrument to image excited state dynamics on a continuous flow device. The experiments presented demonstrate the capability to discriminate between different chromophores as well as in differentiating the effects of local chemical environments that a chromophore experiences. This work presents the first such online transient absorption measurements and provides a new direction for the advancement and optimization of chemical reactions in microfluidic devices.
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The mammalian cerebral cortex is divided into different areas according to their function and pattern of connections. Studies comparing primary visual (V1) and prefrontal cortex (PFC) of primates have demonstrated striking pyramidal neuron (PN) specialization not present in comparable areas of the mouse neocortex. To better understand PFC evolution and regional PN specialization, we studied the tree shrew, a species with a close phylogenetic relationship to primates. We defined the tree shrew PFC based on cytoarchitectonic borders, thalamic connectivity and characterized the morphology and electrophysiology of layer II/III PNs in V1 and PFC. Similar to primates, the PFC PNs in the tree shrew fire with a regular spiking pattern and have larger dendritic tree and spines than those in V1. However, V1 PNs showed strikingly large basal dendritic arbors with high spine density, firing at higher rates and in a more varied pattern than PFC PNs. Yet, unlike in the mouse and unreported in the primate, medial prefrontal PN are more easily recruited than either the dorsolateral or V1 neurons. This specialization of PN morphology and physiology is likely to be a significant factor in the evolution of cortex, contributing to differences in the computational capacities of individual cortical areas.
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Corteza Prefrontal/citología , Corteza Prefrontal/fisiología , Células Piramidales/citología , Células Piramidales/fisiología , Tupaiidae/anatomía & histología , Tupaiidae/fisiología , Corteza Visual/citología , Corteza Visual/fisiología , Animales , Espinas Dendríticas , Femenino , Masculino , Núcleo Talámico Mediodorsal/citología , Potenciales de la Membrana , Vías Visuales/citología , Vías Visuales/fisiologíaRESUMEN
Plasma human immunodeficiency virus type 1 (HIV-1) RNA levels in women are lower early in untreated HIV-1 infection compared with those in men, but women have higher T-cell activation and faster disease progression when adjusted for viral load. It is not known whether these sex differences persist during effective antiretroviral therapy (ART), or whether they would be relevant for the evaluation and implementation of HIV-1 cure strategies. We prospectively enrolled a cohort of reproductive-aged women and matched men on suppressive ART and measured markers of HIV-1 persistence, residual virus activity, and immune activation. The frequency of CD4+ T cells harboring HIV-1 DNA was comparable between the sexes, but there was higher cell-associated HIV-1 RNA, higher plasma HIV-1 (single copy assay), and higher T-cell activation and PD-1 expression in men compared with women. These sex-related differences in immune phenotype and HIV-1 persistence on ART have significant implications for the design and measurement of curative interventions.
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Linfocitos T CD4-Positivos/fisiología , ADN Viral/sangre , Infecciones por VIH/inmunología , VIH-1 , ARN Viral/sangre , Carga Viral , Adulto , Fármacos Anti-VIH/uso terapéutico , Linfocitos T CD4-Positivos/metabolismo , Linfocitos T CD8-positivos/metabolismo , Femenino , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , Humanos , Activación de Linfocitos , Masculino , Persona de Mediana Edad , Receptor de Muerte Celular Programada 1/sangre , Estudios Prospectivos , Receptores CCR5/metabolismo , Factores SexualesRESUMEN
Taylor dispersion analysis (TDA) provides absolute determination of diffusion coefficients for analytes ranging from small molecules to particulate matter. TDA has seen a resurgence in recent years, as modern commercial capillary electrophoresis (CE) instrumentation is well equipped to meet the precision flow requirements of TDA. Discontinuous flow velocities, which occur during sample injection, can lead to substantial inaccuracies in single-point detection TDA. Dual-point detection allows TDA to be carried out under continuous flow in the volume between the detection points, but dual-point fluorescence detection has not previously been feasible within the confines of commercial CE instrumentation. Here, we describe a compact light-emitting diode (LED)-induced fluorescence detector designed for online, dual-point capillary detection within a commercial CE system. The three-dimensional (3D) printed detector houses an inexpensive LED excitation source, a bandpass excitation filter, an integral 3D printed pinhole collimator, and a ball lens, which collects fluorescence emission. Multivariate optimization of operating conditions yielded a detection limit of 613 ± 13 pM for CE of fluorescein disodium salt solution in borate buffer. The miniature size of the device allowed integration of two detectors within a commercial CE system without modification to the instrument, thereby enabling dual-detector assays including TDA and CE-TDA. Monitoring of the bioconjugation reaction between fluorescein isothiocyanate (FITC) and a model protein illustrates the utility of direct, calibration-free size determination, which enabled the resolution of fluorescence originating from free FITC from that of protein-bound FITC. TDA detection coupled to CE enabled the determination of peak identities without the need for standard solutions.
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Black lipid membranes (BLMs) provide a biomimetic model system for studying cellular membrane processes, and are important tools in drug screening and biosensing applications. BLMs offer advantages over liposomes and solid-supported lipid bilayers in applications where access to both leaflets of the bilayer is critical. Reliable and repeatable formation of BLMs presents a major challenge, especially in systems that require interrogation of the membrane via optical microscopy. BLMs for optical interrogation are often formed by the manual painting method, which is tedious and has a high failure rate because it involves manual manipulation of nanoscale liquid films for membrane self-assembly. Here, we describe a fully automated technique for the formation of BLMs within the imaging plane of an inverted fluorescence microscope. The technique utilizes hydrostatic pressure manipulations within a simple microfluidic device, which are feedback controlled via confocal fluorescence monitoring of the BLM formation process. An algorithm for monitoring and precision control of BLM formation is devised and optimized to yield an 80% success rate for the formation of BLMs, with formation times on the order of 78 min. Membranes formed via the automated procedure are confirmed to be fluid and biomimetic via spontaneous insertion of α-hemolysin pores with characteristic conductance of ca. 1 nS. Graphical Abstract á .
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Dispositivos Laboratorio en un Chip , Membrana Dobles de Lípidos , Lípidos de la Membrana/química , Algoritmos , Automatización , Análisis Costo-Beneficio , Presión Hidrostática , Microscopía Confocal , Microscopía Fluorescente , Reproducibilidad de los ResultadosRESUMEN
Contamination of food products by pathogenic microorganisms continues to be a major public health and food industry concern. Non-typhoidal Salmonella species have led to numerous outbreaks associated with various foods. A wide variety of methods have been applied and introduced for treatment of fresh foods to eliminate pathogenic as well as spoilage microorganisms. Salmonella can become exposed to elevated temperatures while associated with hosts such as poultry. In addition, heat treatment is also applied at various stages of processing to retain the shelf life of food products. Despite this, these microorganisms may overcome exposure to such treatments through the efficient expression of stress response mechanisms and result in illness following consumption. Thermal stress induces a range of destructive exposures to bacterial cells such as protein damage and DNA damage caused by reactive oxygen species. In this study, we chose three genes (∆recD, ∆STM14_5307, and ∆aroD) associated with conditionally essential genes required for different aspects of optimal growth at 42 °C and evaluated the responses of wild type and mutant Salmonella Typhimurium strains to uncover potential mechanisms that may enable survival and resistance under thermal stress. The RecBCD complex that initiates repair of double-stranded DNA breaks through homologous recombination. STM14_5307 is a transcriptional regulator involved in stationary phase growth and inositol metabolism. The gene aroD is involved in metabolism and stationary phase growth. These strains were characterized via high throughput phenotypic profiling in response to two different growth temperatures (37 °C (human host temperature) and 42 °C (poultry host temperature)). The ∆aroD strain exhibited the highest sensitivity to the various temperatures followed by the ∆recD and ∆STM14_5307 strains, respectively. Achieving more understanding of the molecular mechanisms of heat survival may lead to the development of more effective strategies to limit Salmonella in food products through thermal treatment by developing interventions that specifically target the pathways these genes are involved in.
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Viabilidad Microbiana/efectos de la radiación , Fenotipo , Salmonella typhimurium/fisiología , Salmonella typhimurium/efectos de la radiación , Temperatura , Proteínas Bacterianas/genética , Exodesoxirribonucleasa V/genética , Eliminación de Gen , Genes Reguladores , Humanos , Salmonella typhimurium/genética , Estrés FisiológicoRESUMEN
Certain pathogenic Escherichia coli known as Shiga toxin (Stx)-producing E. coli (STEC) are a public health threat to the consumer, and are problematic for the food industry. Food products containing STEC are deemed unfit for human consumption, and STEC illnesses can cause hemolytic uremic syndrome (HUS), a disease affecting the kidneys in susceptible individuals. Optimizing detection methods in foods have been focused on more prompt and accurate analysis. This review addresses the role and applications of immuno-based assays for STEC detection in food systems. Immunoassay antibody capture systems and flow cytometry platforms have been implemented into several food-based detection systems. By applying antibodies that will interact with target microorganisms, immunoassays can be used to directly detect and quantify pathogens. Immuno-based protocols could potentially be further implemented into the food industry, limit the duration of the detection process and increase accuracy.
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Microbiología de Alimentos/métodos , Inmunoensayo/métodos , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Citometría de Flujo/métodosRESUMEN
Salmonella serovars, one of the leading contributors to foodborne illness and are especially problematic for foods that are not cooked before consumption, such as fresh produce. The shipping containers that are used to transport and store fresh produce may play a role in cross contamination and subsequent illnesses. However, methods for quantitatively attached cells are somewhat variable. The overall goal of this study was to compare conventional plating with molecular methods for quantitating attached representative strains for Salmonella Typhimurium and Heidelberg on reusable plastic containers (RPC) coupons, respectively. We attached Salmonella enterica serovar Typhimurium ATCC 14028 and serovar Heidelberg SL486 (parent and an antibiotic resistant marker strain) to plastic coupons (2.54 cm(2)) derived from previously used shipping containers by growing for 72 h in tryptic soy broth. The impact of the concentration of sanitizer on log reductions between unsanitized and sanitized coupons was evaluated by exposing attached S. Typhimurium cells to 200 ppm and 200,000 ppm sodium hypochlorite (NaClO). Differences in sanitizer effectiveness between serovars were also evaluated with attached S. Typhimurium compared to attached S. Heidelberg populations after being exposed to 200 ppm peracetic acid (PAA). Treatment with NaClO caused an average of 2.73 ± 0.23 log CFU of S. Typhimurium per coupon removed with treatment at 200 ppm while 3.36 ± 0.54 log CFU were removed at 200,000 ppm. Treatment with PAA caused an average of 2.62 ± 0.15 log CFU removed for S. Typhimurium and 1.41 ± 0.17 log CFU for S. Heidelberg (parent) and 1.61 ± 0.08 log CFU (marker). Lastly, scanning electron microscopy (SEM) was used to visualize cell attachment and coupon surface topography. SEM images showed that remaining attached cell populations were visible even after sanitizer application. Conventional plating and qPCR yielded similar levels of enumerated bacterial populations indicating a high concordance between the two methods. Therefore, qPCR could be used for the rapid quantification of Salmonella attached on RPC.
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Técnicas de Tipificación Bacteriana/métodos , Desinfectantes , Embalaje de Alimentos , Inocuidad de los Alimentos , Plásticos , Salmonella enterica/clasificación , Salmonella enterica/aislamiento & purificación , Recuento de Colonia Microbiana , Salmonella typhimurium/clasificación , Salmonella typhimurium/aislamiento & purificaciónRESUMEN
The purpose of this study was to characterize the microbial community in ground pork using molecular approaches. Forty six ground pork products were purchased from local stores in the north central area of South Korea. Aerobic plate counts varied 4.23 ± 5.14 × 10(5) CFU/g with the range between 5.00 × 10(3) and 1.85 × 10(6) CFU/g for ground pork samples. Four ground meat samples were further processed for metagenomic analysis. Pseudomonas species was the most relative abundant with a wide range occurring (1.72 to 77.7%) as part of the microbial genera in ground pork. Bacteria such as Carnobacterium, Yersinia, Photobacterium were also identified in ground pork. Despite the prominence of certain genera across all samples there was still extensive microbial diversity among ground pork products that originated from different slaughter houses and were processed in different markets. Such diversity indicates that designing interventions to extend shelf life may be hampered by the extensive variability in the microbial consortia associated with pork products. However, this diversity may be useful for developing microbial traceability signatures unique to a slaughter house or a particular market.
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Microbiología de Alimentos , Microbiota , Carne Roja/microbiología , Animales , Servicios de Alimentación , Metagenómica , República de Corea , PorcinosRESUMEN
Tatsoi (Brassica narinosa) is a popular Asian salad green that is mostly consumed as a source of fresh produce. The purpose of this study was to assess the microbial diversity of Tatsoi cultivated in a hydroponic system and of its ecosystem. Tatsoi leaves, nutrient solution, and perlite/earth samples from a trickle feed system (TFS) and an ebb-and-flow system (EFS) were collected and their microbial communities were analyzed by pyrosequencing analysis. The results showed that most bacteria in the leaves from the TFS contained genus Sporosarcina (99.6%), while Rhizobium (60.4%) was dominant in the leaves from the EFS. Genus Paucibacter (18.21%) and Pelomonas (12.37%) were the most abundant microbiota in the nutrient solution samples of the TFS. In the EFS, the nutrient solution samples contained mostly genus Rhodococcus and Acinetobacter. Potential microbial transfer between the leaves and the ecosystem was observed in the EFS, while samples in the TFS were found to share only one species between the leaves, nutrient solution, and earth. Together, these results show that the bacterial populations in Tatsoi and in its ecosystem are highly diverse based on the cultivation system.
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Bacterias , Brassica/microbiología , Microbiología de Alimentos , Hidroponía , Microbiota , Hojas de la Planta/microbiología , Verduras/microbiología , Bacterias/clasificación , Bacterias/aislamiento & purificaciónRESUMEN
Suspended lipid membranes, also called black lipid membranes (BLMs), are an important model system that approximates the lipid bilayer environment of cell membranes. Increasingly, BLMs are utilized in sensing strategies that harness high sensitivity measurements of ion flux across the membrane, typically facilitated by ion channel proteins. BLMs are suspended across microapertures that connect two otherwise isolated fluidic compartments, and the precision fabrication of such microapertures can contribute to the stability and performance of the resulting BLM. Here, we highlight two emerging trends in the precision fabrication of microapertures for BLM formation: microfabrication in silicon-based thin film substrates, and microfabrication in the negative photoresist material SU-8. Four unique fabrication strategies are outlined, and we project the impact that these microfabrication strategies will have for BLM-integrated bioanalytical technologies.
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Bioquímica/instrumentación , Membrana Dobles de Lípidos , Microtecnología/métodos , Compuestos Epoxi/química , Diseño de Equipo , Lípidos de la Membrana , Polímeros/química , SilicioRESUMEN
Salmonella enterica is one of the most prevalent pathogens responsible for foodborne illness worldwide. Numerous Salmonella serovars have been associated with the consumption of a variety of products, and limiting food-borne illness due to Salmonella serovars is a continuing problem for food producers and public health. The emergence and prevalence of Salmonella serovars has been studied but the predominant serovars have varied somewhat over the years. The aims of this research were to compare the aerobic growth responses of selected predominant foodborne Salmonella serovars, and evaluate how the spent media from different serovars affects the growth of a well-characterized Salmonella Typhimurium strain. Growth responses were similar for most strains in spent media except for S. Typhimurium (ATCC 14028), which exhibited a decrease in growth in the presence of Salmonella Heidelberg (ARI-14) spent media. This research will provide a better understanding of the growth differences among several Salmonella serovars in nutrient limited spent media.