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1.
Rev Sci Instrum ; 95(2)2024 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-38391285

RESUMO

A synthetic Mirnov diagnostic has been developed to investigate the capabilities and limitations of an arrangement of Mirnov coils in terms of a mode analysis. Eight test cases have been developed, with different coil arrangements and magnetic field configurations. Three of those cases are experimental configurations of the stellarator Wendelstein 7-X. It is observed that, for a high triangularity of the flux surfaces, the arrangement of the coils plays a significant role in the exact determination of the poloidal mode number. For the mode analysis, torus and magnetic coordinates have been used. In most cases, the reconstruction of the poloidal mode number of a prescribed mode was found to be more accurate in magnetic coordinates. As an application, the signal of an Alfvén eigenmode, which has been calculated with a three-dimensional magnetohydrodynamics code, is compared to experimental observations at Wendelstein 7-X. For the chosen example, the calculated and measured mode spectra agree very well and additional information on the toroidal mode number and localization of the mode has been inferred.

3.
Nature ; 596(7871): 221-226, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34381232

RESUMO

Research on magnetic confinement of high-temperature plasmas has the ultimate goal of harnessing nuclear fusion for the production of electricity. Although the tokamak1 is the leading toroidal magnetic-confinement concept, it is not without shortcomings and the fusion community has therefore also pursued alternative concepts such as the stellarator. Unlike axisymmetric tokamaks, stellarators possess a three-dimensional (3D) magnetic field geometry. The availability of this additional dimension opens up an extensive configuration space for computational optimization of both the field geometry itself and the current-carrying coils that produce it. Such an optimization was undertaken in designing Wendelstein 7-X (W7-X)2, a large helical-axis advanced stellarator (HELIAS), which began operation in 2015 at Greifswald, Germany. A major drawback of 3D magnetic field geometry, however, is that it introduces a strong temperature dependence into the stellarator's non-turbulent 'neoclassical' energy transport. Indeed, such energy losses will become prohibitive in high-temperature reactor plasmas unless a strong reduction of the geometrical factor associated with this transport can be achieved; such a reduction was therefore a principal goal of the design of W7-X. In spite of the modest heating power currently available, W7-X has already been able to achieve high-temperature plasma conditions during its 2017 and 2018 experimental campaigns, producing record values of the fusion triple product for such stellarator plasmas3,4. The triple product of plasma density, ion temperature and energy confinement time is used in fusion research as a figure of merit, as it must attain a certain threshold value before net-energy-producing operation of a reactor becomes possible1,5. Here we demonstrate that such record values provide evidence for reduced neoclassical energy transport in W7-X, as the plasma profiles that produced these results could not have been obtained in stellarators lacking a comparably high level of neoclassical optimization.

4.
J Dairy Sci ; 102(7): 6682-6698, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31128869

RESUMO

Our objective was to compare the composition of bedding materials and manure, cow welfare and hygiene assessments, measures of milk production and quality, and incidence of mastitis during a 3-yr trial with lactating Holstein cows housed in a freestall barn containing 4 identical pens with 32 freestalls/pen. Bedding systems evaluated included deep-bedded organic manure solids (DBOS), shallow-bedded manure solids spread over mattresses (MAT), deep-bedded recycled sand (RSA), and deep-bedded new sand (NSA). The experiment was designed as a 4 × 4 Latin square with 4 bedding systems and 4 experimental periods, but was terminated after 3 yr following discussions with the consulting statistician; therefore, data were analyzed as an incomplete Latin square. A total of n = 734 mostly primiparous cows (n = 725 primiparous, n = 9 multiparous; 224 to 267 cows/yr) were enrolled in the trial. Before placement in freestalls, organic solids (OS) exhibited lower concentrations of dry matter (36.5 vs. 94.3%), and greater concentrations of volatile solids, C, N, NH4-N, P, water-extractable P, K, and S compared with RSA or NSA. Cow comfort index was greater for sand-bedded systems compared with those using OS (88.4 vs. 82.8%). Cows bedded in systems using OS (DBOS and MAT) exhibited greater mean hock scores (1 = no swelling, no hair loss; 2 = no swelling, bald area on hock) than those bedded in sand (1.25 vs. 1.04), but this effect was entirely associated with use of mattresses (MAT), which differed sharply from DBOS (1.42 vs. 1.07). Generally, hygiene scores for legs, flanks, and udders were numerically similar for DBOS, NSA, and RSA bedding systems, and differences between bedding systems were associated entirely with MAT, yielding detectable contrasts between MAT and DBOS for legs (2.94 vs. 2.20), flanks (2.34 vs. 1.68), and udders (1.83 vs. 1.38). No significant contrast comparing bedding systems was detected for measures of milk production or quality. Documented cases of clinical mastitis requiring treatment ranged from a low rate of 7.4 cases/yr for RSA to a high of 23.1 cases/yr for DBOS, based on a mean enrollment of 60.7 to 63.0 cows/treatment per yr. Cows bedded with OS exhibited a greater incidence of mastitis than those bedded with sand (19.0 vs. 8.4 cases/yr), but no differences were observed for comparisons within individual bedding-material types. Collectively, these results generally favored use of sand-bedding materials over systems using OS.


Assuntos
Bovinos , Indústria de Laticínios/métodos , Abrigo para Animais , Dióxido de Silício , Criação de Animais Domésticos , Animais , Feminino , Higiene , Incidência , Lactação , Mastite Bovina/epidemiologia , Mastite Bovina/prevenção & controle , Distribuição Aleatória
5.
Water Res ; 157: 356-364, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30970285

RESUMO

Enteric viruses pose the greatest acute human health risks associated with subsurface drinking water supplies, yet quantitative risk assessment tools have rarely been used to develop health-based targets for virus treatment in drinking water sourced from these supplies. Such efforts have previously been hampered by a lack of consensus concerning a suitable viral reference pathogen and dose-response model as well as difficulties in quantifying pathogenic viruses in water. A reverse quantitative microbial risk assessment (QMRA) framework and quantitative polymerase chain reaction data for norovirus genogroup I in subsurface drinking water supplies were used herein to evaluate treatment needs for such water supplies. Norovirus was not detected in over 90% of samples, which emphasizes the need to consider the spatially and/or temporally intermittent patterns of enteric pathogen contamination in subsurface water supplies. Collectively, this analysis reinforces existing recommendations that a minimum 4-log treatment goal is needed for enteric viruses in groundwater in absence of well-specific monitoring information. This result is sensitive to the virus dose-response model used as there is approximately a 3-log discrepancy among virus dose-response models in the existing literature. This emphasizes the need to address the uncertainties and lack of consensus related to various QMRA modelling approaches and the analytical limitations that preclude more accurate description of virus risks.


Assuntos
Água Potável , Água Subterrânea , Vírus , Desinfecção , Humanos , Microbiologia da Água , Abastecimento de Água
6.
J Anim Sci ; 96(3): 964-974, 2018 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-29401268

RESUMO

Dairy slurry is used commonly as an animal-sourced fertilizer in agronomic production. However, residual effects of slurry application on intake and digestibility of alfalfa (Medicago sativa L.) silage from subsequent harvests are not well known. The objective of this study was to determine if moisture concentration of alfalfa silage and timing of dairy slurry application relative to subsequent harvest affected intake and digestibility by sheep. Katahdin crossbred ewes (n = 18; 48 ± 5.3 kg) in mid-gestation were stratified by BW and allocated randomly in each of two periods to one of six treatments arranged in a two × three factorial arrangement. Treatments consisted of recommended (RM; 46.8%) or low (LM; 39.7%) moisture at baling after either no slurry application (NS), slurry application to stubble immediately after removal of the previous cutting (S0), or slurry application 14 d after removal of the previous cutting (S14). Silages were chopped through a commercial straw chopper, packed into plastic trash cans, and then offered to ewes within 4 d of chopping. Period 1 of the intake and digestion study consisted of a 14-d adaptation followed by a 7-d fecal collection period. Period 2 followed period 1 after a 4-d rest and consisted of an 11-d adaptation followed by 7 d of fecal collection. Ewes were housed individually in 1.4 × 4.3-m pens equipped with rubber mat flooring. Feces were swept from the floor twice daily, weighed, and dried at 50 °C. Ewes had ad libitum access to water and were offered chopped silage for a minimum of 10% refusal (DM). Blood samples were collected immediately prior to feeding, and 4 and 8 h after feeding on the day prior to the end of each period. Organic matter intake (g/kg BW) and OM digestibility tended (P < 0.10) to be, and digestible OM intake (g/kg BW) was reduced by slurry application. Lymphocytes (% of total white blood cells) were greater (P < 0.05) from LM vs. RM and from NS vs. S0 and S14. Red blood cell concentrations were greater (P < 0.05) from S14 vs. S0 and from S0 and S14 vs. NS. Serum urea N concentrations did not differ (P > 0.17) across treatments. Therefore, moisture concentration of alfalfa silage within the range used in this study may not affect voluntary intake or digestibility, but slurry application may have an effect on digestible OM intake. Also, moisture concentration of alfalfa silage and time of dairy slurry application may affect specific blood hemograms.


Assuntos
Fertilizantes/análise , Medicago sativa , Ovinos/fisiologia , Silagem/análise , Animais , Nitrogênio da Ureia Sanguínea , Dieta/veterinária , Digestão , Fezes , Feminino , Fermentação , Fertilizantes/efeitos adversos , Esterco , Distribuição Aleatória
7.
Water Res ; 113: 11-21, 2017 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-28187346

RESUMO

Great Lakes tributaries are known to deliver waterborne pathogens from a host of sources. To examine the hydrologic, land cover, and seasonal patterns of waterborne pathogens (i.e. protozoa (2), pathogenic bacteria (4) human viruses, (8) and bovine viruses (8)) eight rivers were monitored in the Great Lakes Basin over 29 months from February 2011 to June 2013. Sampling locations represented a wide variety of land cover classes from urban to agriculture to forest. A custom automated pathogen sampler was deployed at eight sampling locations which provided unattended, flow-weighted, large-volume (120-1630 L) sampling. Human and bovine viruses and pathogenic bacteria were detected by real-time qPCR in 16%, 14%, and 1.4% of 290 samples collected while protozoa were never detected. The most frequently detected pathogens were: bovine polyomavirus (11%), and human adenovirus C, D, F (9%). Human and bovine viruses were present in 16.9% and 14.8% of runoff-event samples (n = 189) resulting from precipitation and snowmelt, and 13.9% and 12.9% of low-flow samples (n = 101), respectively, indicating multiple delivery mechanisms could be influential. Data indicated human and bovine virus prevalence was different depending on land cover within the watershed. Occurrence, concentration, and flux of human viruses were greatest in samples from the three sampling locations with greater than 25% urban influence than those with less than 25% urban influence. Similarly, occurrence, concentration, and flux of bovine viruses were greatest in samples from the two sampling locations with greater than 50 cattle/km2 than those with less than 50 cattle/km2. In seasonal analysis, human and bovine viruses occurred more frequently in spring and winter seasons than during the fall and summer. Concentration, occurrence, and flux in the context of hydrologic condition, seasonality, and land use must be considered for each watershed individually to develop effective watershed management strategies for pathogen reduction.


Assuntos
Lagos , Estações do Ano , Animais , Bovinos , Monitoramento Ambiental , Humanos , Hidrologia , Rios
8.
Rev Sci Instrum ; 87(11): 11E511, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27910553

RESUMO

The Thomson scattering diagnostic on the Madison Symmetric Torus (MST) records excessive levels of stray Nd:YAG laser light. Stray light saturates the 1064 nm spectral channel in all polychromators, which prevents absolute electron density measurements via Rayleigh scattering calibration. Furthermore, stray light contaminates adjacent spectral channels for r/a ≥ 0.75, which renders the diagnostic unable to make electron temperature measurements at these radii. In situ measurements of stray light levels during a vacuum vessel vent are used to identify stray light sources and strategies for reduction of stray light levels. Numerical modeling using Zemax OpticStudio supports these measurements. The model of the vacuum vessel and diagnostic includes synthetic collection optics to enable direct comparison of measured and simulated stray light levels. Modeling produces qualitatively similar stray light distributions to MST measurements, and quantifies the mitigation effects of stray light mitigation strategies prior to implementation.

9.
Rev Sci Instrum ; 87(11): 11E547, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27910649

RESUMO

The Thomson scattering diagnostic on MST records both equilibrium and fluctuating electron temperature with a range capability of 10 eV-5 keV. Standard operation with two modified commercial Nd:YAG lasers allows measurements at rates of 1 kHz-25 kHz. Several subsystems of the diagnostic are being improved. The power supplies for the avalanche photodiode detectors (APDs) that record the scattered light are being replaced to improve usability, reliability, and maintainability. Each of the 144 APDs will have an individual rack mounted switching supply, with bias voltage adjustable to match the APD. Long-wavelength filters (1140 nm center, 80 nm bandwidth) have been added to the polychromators to improve capability to resolve non-Maxwellian distributions and to enable directed electron flow measurements. A supercontinuum (SC) pulsed white light source has replaced the tungsten halogen lamp previously used for spectral calibration of the polychromators. The SC source combines substantial brightness produced in nanosecond pulses with a spectrum that covers the entire range of the polychromators.

10.
Sportverletz Sportschaden ; 28(4): 204-10, 2014 Dec.
Artigo em Alemão | MEDLINE | ID: mdl-25423198

RESUMO

INTRODUCTION: The growing numbers of members aged over 35 years in the German Dancesport Association indicate that not only physical activity but also sporting success is gaining significance with increasing age. Investigations on health hazards are still lacking. Aim of this study is the analysis of dance sport-related health hazards in the classifications Seniors I-III. METHODS: A total of n = 124 (m: n = 67, f: n = 57) senior ballroom dancers participated in this retrospective cross-sectional investigation. RESULTS: There were 0.9 (m)/1.0 (f) traumatic injuries/year (m: 0.03/1000 h, f: 0.04/1000 h). Gender specific differences as to localisation, type and factors could be observed. The most common injury localisations were the spine (22.2 %), followed by upper and lower leg (15.9 %) in males with foot (incl. ankle joint) (35.6 %), followed by spine (25.4 %), hip (15.3 %) and knee joint (23.8 %) in females. Chronic sports damages/complaints were more common in dancers (m: 1.6, f: 1.9) than traumatic injuries (m: 0.05/1000 h, w: 007/1000 h). The number of traumatic injuries and chronic sports damages rose according to the extent of training and age in males, with females only according to age. Knee problems (arthrosis/gonalgia/meniscal damage) were the most common sports damages (m: 30 %, f: 19.4 %), followed by degenerative spine diseases (m: 9.1 %, f: 9.7 %) and complaints not yet diagnosed (m: 14.6 %, f: 16 %). Intrinsic factors predominated (m: 64.7 %, f: 53.6 %). CONCLUSION/DISCUSSION: The injury risk in competitive senior ballroom dancers is low. An increase of jeopardising with age and extent of training could only be observed in males. That raises the issue of an optimal and gender-specific amount of training and the preventive significance of dance sport in the elderly. Localisation and type of injury reflect the characteristic movement elements in dance sport. There is a need for additional qualitative and quantitative investigations in order to create differentiated suggestions as to the planning of training and injury prevention.


Assuntos
Traumatismos do Braço/epidemiologia , Traumatismos em Atletas/epidemiologia , Dança/lesões , Dança/estatística & dados numéricos , Traumatismos da Perna/epidemiologia , Osteoartrite do Joelho/epidemiologia , Adulto , Distribuição por Idade , Idoso , Comorbidade , Feminino , Alemanha/epidemiologia , Pesquisas sobre Atenção à Saúde , Humanos , Incidência , Degeneração do Disco Intervertebral/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Distribuição por Sexo
11.
J Dairy Sci ; 97(11): 7197-211, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25242431

RESUMO

Dairy producers frequently ask questions about the risks associated with applying dairy slurry to growing alfalfa (Medicago sativa L.). Our objectives were to determine the effects of applying dairy slurry on the subsequent nutritive value and fermentation characteristics of alfalfa balage. Dairy slurry was applied to 0.17-ha plots of alfalfa; applications were made to the second (HARV1) and third (HARV2) cuttings during June and July of 2012, respectively, at mean rates of 42,400 ± 5271 and 41,700 ± 2397 L/ha, respectively. Application strategies included (1) no slurry, (2) slurry applied directly to stubble immediately after the preceding harvest, (3) slurry applied after 1 wk of post-ensiled regrowth, or (4) slurry applied after 2 wk of regrowth. All harvested forage was packaged in large, rectangular bales that were ensiled as wrapped balage. Yields of DM harvested from HARV1 (2,477 kg/ha) and HARV2 (781 kg/ha) were not affected by slurry application treatment. By May 2013, all silages appeared to be well preserved, with no indication of undesirable odors characteristic of clostridial fermentations. Clostridium tyrobutyricum, which is known to negatively affect cheese production, was not detected in any forage on either a pre- or post-ensiled basis. On a pre-ensiled basis, counts for Clostridium cluster 1 were greater for slurry-applied plots than for those receiving no slurry, and this response was consistent for HARV1 (4.44 vs. 3.29 log10 genomic copies/g) and HARV2 (4.99 vs. 3.88 log10 genomic copies/g). Similar responses were observed on a post-ensiled basis; however, post-ensiled counts also were greater for HARV1 (5.51 vs. 5.17 log10 genomic copies/g) and HARV2 (5.84 vs. 5.28 log10 genomic copies/g) when slurry was applied to regrowth compared with stubble. For HARV2, counts also were greater following a 2-wk application delay compared with a 1-wk delay (6.23 vs. 5.45 log10 genomic copies/g). These results suggest that the risk of clostridial fermentations in alfalfa silages is greater following applications of slurry. Based on pre- and post-ensiled clostridial counts, applications of dairy slurry on stubble are preferred (and less risky) compared with delayed applications on growing alfalfa.


Assuntos
Fertilizantes/análise , Medicago sativa/metabolismo , Valor Nutritivo , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Fermentação , Fertilizantes/efeitos adversos , Esterco , Medicago sativa/química , Silagem/análise
12.
Sci Total Environ ; 490: 849-60, 2014 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-24908645

RESUMO

To examine the occurrence, hydrologic variability, and seasonal variability of human and bovine viruses in surface water, three stream locations were monitored in the Milwaukee River watershed in Wisconsin, USA, from February 2007 through June 2008. Monitoring sites included an urban subwatershed, a rural subwatershed, and the Milwaukee River at the mouth. To collect samples that characterize variability throughout changing hydrologic periods, a process control system was developed for unattended, large-volume (56-2800 L) filtration over extended durations. This system provided flow-weighted mean concentrations during runoff and extended (24-h) low-flow periods. Human viruses and bovine viruses were detected by real-time qPCR in 49% and 41% of samples (n=63), respectively. All human viruses analyzed were detected at least once including adenovirus (40% of samples), GI norovirus (10%), enterovirus (8%), rotavirus (6%), GII norovirus (1.6%) and hepatitis A virus (1.6%). Three of seven bovine viruses analyzed were detected including bovine polyomavirus (32%), bovine rotavirus (19%), and bovine viral diarrhea virus type 1 (5%). Human viruses were present in 63% of runoff samples resulting from precipitation and snowmelt, and 20% of low-flow samples. Maximum human virus concentrations exceeded 300 genomic copies/L. Bovine viruses were present in 46% of runoff samples resulting from precipitation and snowmelt and 14% of low-flow samples. The maximum bovine virus concentration was 11 genomic copies/L. Statistical modeling indicated that stream flow, precipitation, and season explained the variability of human viruses in the watershed, and hydrologic condition (runoff event or low-flow) and season explained the variability of the sum of human and bovine viruses; however, no model was identified that could explain the variability of bovine viruses alone. Understanding the factors that affect virus fate and transport in rivers will aid watershed management for minimizing human exposure and disease transmission.


Assuntos
Monitoramento Ambiental , Rios/virologia , Vírus/crescimento & desenvolvimento , Microbiologia da Água , Animais , Bovinos , Humanos , Vírus/classificação , Wisconsin
13.
Rev Sci Instrum ; 83(10): 10E324, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23126982

RESUMO

Calibration of the Madison Symmetric Torus Thomson scattering system has been refined to improve temperature fluctuation measurements. Multiple avalanche photodiodes have been directly calibrated for use as reference detectors during calibration, improving accuracy and ease of use. From the absolute calibration we calculate corrections to the gain for variation in detector operating temperature. We also measure the spatial uniformity of detector responsivity for several photodiodes, and present a method of accounting for non-uniformity in the calibration process. Finally, the gain and noise enhancement are measured at multiple wavelengths to improve temperature and uncertainty measurements.

14.
Water Res ; 46(13): 4281-91, 2012 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-22673345

RESUMO

Naturally-occurring inhibitory compounds are a major concern during qPCR and RT-qPCR analysis of environmental samples, particularly large volume water samples. Here, a standardized method for measuring and mitigating sample inhibition in environmental water concentrates is described. Specifically, the method 1) employs a commercially available standard RNA control; 2) defines inhibition by the change in the quantification cycle (C(q)) of the standard RNA control when added to the sample concentrate; and 3) calculates a dilution factor using a mathematical formula applied to the change in C(q) to indicate the specific volume of nuclease-free water necessary to dilute the effect of inhibitors. The standardized inhibition method was applied to 3,193 large-volume water (surface, groundwater, drinking water, agricultural runoff, sewage) concentrates of which 1,074 (34%) were inhibited. Inhibition level was not related to sample volume. Samples collected from the same locations over a one to two year period had widely variable inhibition levels. The proportion of samples that could have been reported as false negatives if inhibition had not been mitigated was between 0.3% and 71%, depending on water source. These findings emphasize the importance of measuring and mitigating inhibition when reporting qPCR results for viral pathogens in environmental waters to minimize the likelihood of reporting false negatives and under-quantifying virus concentration.


Assuntos
Vírus GB C/genética , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Microbiologia da Água , Água Potável/virologia , Monitoramento Ambiental/métodos , Vírus GB C/isolamento & purificação , Água Subterrânea/virologia , RNA Viral/isolamento & purificação , Reprodutibilidade dos Testes , Esgotos/virologia
15.
Rev Sci Instrum ; 81(10): 10D513, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21033868

RESUMO

Two standard commercial flashlamp-pumped Nd:YAG (YAG denotes yttrium aluminum garnet) lasers have been upgraded to "pulse-burst" capability. Each laser produces a burst of up to 15 2 J Q-switched pulses (1064 nm) at repetition rates of 1-12.5 kHz. Variable pulse-width drive (0.15-0.39 ms) of the flashlamps is accomplished by insulated gate bipolar transistor (IGBT) switching of electrolytic capacitor banks. Direct control of the laser Pockels cell drive enables optimal pulse energy extraction, and up to four 2 J laser pulses during one flashlamp pulse. These lasers are used in the Thomson scattering plasma diagnostic system on the MST reversed-field pinch to record the dynamic evolution of the electron temperature profile and temperature fluctuations. To further these investigations, a custom pulse-burst laser system with a maximum pulse repetition rate of 250 kHz is now being commissioned.

16.
J Virol Methods ; 163(2): 244-52, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19835913

RESUMO

Quantifying infectious viruses by cell culture depends on visualizing cytopathic effect, or for integrated cell culture-PCR, attaining confidence a PCR-positive signal is the result of virus growth and not inoculum carryover. This study developed mathematical methods to calculate infectious virus numbers based on viral growth kinetics in cell culture. Poliovirus was inoculated into BGM cell monolayers at 10 concentrations from 0.001 to 1000 PFU/ml. Copy numbers of negative-strand RNA, a marker of infectivity for single-stranded positive RNA viruses, were measured over time by qRT-PCR. Growth data were analyzed by two approaches. First, data were fit with a continuous function to estimate directly the initial virus number, expressed as genomic copies. Such estimates correlated with actual inoculum numbers across all concentrations (R(2)=0.62, n=17). Second, the length of lag phase appeared to vary inversely with inoculum titers; hence, standard curves to predict inoculum virus numbers were derived based on three definitions of lag time: (1) time of first detection of (-)RNA, (2) second derivative maximum of the fitted continuous function, and (3) time when the fitted curve crossed a threshold (-)RNA concentration. All three proxies yielded standard curves with R(2)=0.69-0.90 (n=17). The primary advantage of these growth kinetics approaches is being able to quantify virions that are unambiguously infectious, a particular advantage for viruses that do not produce CPE.


Assuntos
Microbiologia Ambiental , Modelos Teóricos , Reação em Cadeia da Polimerase/métodos , Carga Viral , Virologia/métodos , Vírus/crescimento & desenvolvimento , Animais , Técnicas de Cultura de Células , Chlorocebus aethiops , Dosagem de Genes , Vírus/genética
17.
J Appl Microbiol ; 107(4): 1089-97, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19486387

RESUMO

AIMS: To evaluate the effectiveness of continuous separation channel centrifugation for concentrating Toxoplasma gondii and Cyclospora cayetanensis from drinking water and environmental waters. METHODS AND RESULTS: Ready-to-seed vials with known quantities of T. gondii and C. cayetanensis oocysts were prepared by flow cytometry. Oocysts were seeded at densities ranging from 1 to 1000 oocysts l(-1) into 10 to 100 l test volumes of finished drinking water, water with manipulated turbidity, and the source waters from nine drinking water utilities. Oocysts were recovered using continuous separation channel centrifugation and counted on membrane filters using epifluorescent microscopy. Recovery efficiencies of both parasites were > or =84% in 10 l volumes of drinking water. In source waters, recoveries ranged from 64% to 100%, with the lowest recoveries in the most turbid waters. Method precision was between 10% and 20% coefficient of variation. CONCLUSION: Toxoplasma gondii and C. cayetanensis are effectively concentrated from various water matrices by continuous separation channel centrifugation. SIGNIFICANCE AND IMPACT OF THE STUDY: Waterborne transmission of T. gondii and C. cayetanensis presents another challenge in producing clean drinking water and protecting public health. Detection of these parasites relies on effectively concentrating oocysts from ambient water, otherwise false negatives may result. Validation data specific to T. gondii and C. cayetanensis concentration methods are limited. Continuous separation channel centrifugation recovers oocysts with high efficiency and precision, the method attributes required to accurately assess the risk of waterborne transmission.


Assuntos
Centrifugação/métodos , Cyclospora/isolamento & purificação , Água Doce/parasitologia , Toxoplasma/isolamento & purificação , Microbiologia da Água , Abastecimento de Água , Animais , Monitoramento Ambiental , Oocistos/parasitologia , Abastecimento de Água/normas
18.
Rev Sci Instrum ; 79(10): 10E733, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19044549

RESUMO

The multipoint Thomson scattering diagnostic on the Madison Symmetric Torus (MST) is now fully operational with 21 spatial points, which cover the entire minor radius. Four full electron temperature profiles can be obtained during each MST discharge, with a variable delay between each profile. This system overcomes challenges that arise from the unique machine design, location, and plasma characteristics of MST. The machine design limits the maximum porthole diameter to 11.4 cm, requiring a compact, re-entrant, seven element lens for scattered light collection. Limited space near MST necessitates a long beam path for the two Nd:YAG lasers requiring a remote beam line adjustment system to suppress drift in the beam position due to thermal expansion of the building. Due to the remote location of the laser head, substantial design effort was put into the creation of a set of safety interlocks for the laser system. The dynamic nature of MST plasmas and the wide range of operating space require a versatile scattered light detection system consisting of filter polychromators with temperature controlled avalanche photodiode detectors. We also implement an insertable integrating sphere, which travels along the laser beam path through the vacuum vessel, for the alignment of both the fiber optics and the lasers.

19.
Rev Sci Instrum ; 79(10): 10E734, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19044550

RESUMO

Detailed calibrations of the Madison Symmetric Torus polychromator Thomson scattering system have been made suitable for electron temperature fluctuation measurements. All calibrations have taken place focusing on accuracy, ease of use and repeatability, and in situ measurements wherever possible. Novel calibration processes have been made possible with an insertable integrating sphere (ISIS), using an avalanche photodiode (APD) as a reference detector and optical parametric oscillator (OPO). Discussed are a novel in situ spatial calibration with the use of the ISIS, the use of an APD as a reference detector to streamline the APD calibration process, a standard dc spectral calibration, and in situ pulsed spectral calibration made possible with a combination of an OPO as a light source, the ISIS, and an APD used as a reference detector. In addition a relative quantum efficiency curve for the APDs is obtained to aid in uncertainty analysis.

20.
Rev Sci Instrum ; 79(10): 10E735, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19044551

RESUMO

The Madison Symmetric Torus (MST) presents challenging conditions for Thomson scattering (TS) measurements. The MST plasmas are reversed-field pinches (RFPs) with electron density n(e)<3x10(13) cm(-3), typically 1x10(13) cm(-3). The TS system was designed to measure from 10 eV to 2 keV; however, six polychromators were upgraded from four to eight spectral channels to resolve to 10 keV. There is no diverter or vertical field, so wall interaction results in high background light both from ion and neutral bremsstrahlungs and from line radiation. Also during standard plasmas, the RFP exhibits regular reconnection sawteeth events during which the plasma current, density, and temperature profiles are flattened. These events are of interest both due to the reconnection physics and to their contribution to the MST equilibrium and confinement. These events occur over 100 microS and exhibit large changes in background light and fast changes in temperature. During improved confinement plasmas, there are no sawteeth; the background is low but the temperature can be over an order of magnitude higher. Data analysis of the system has been developed to accommodate both the large dynamic range of the temperature, the fast dynamics, and the fast changing, high amplitude background. Special attention has been paid to the sources of error, in particular, the contribution of the background. A response-function method reduces the measured uncertainty by a factor of 2. Numerical techniques have been developed which are extremely robust. Two methods are used, a conventional chi(2) minimization using a Levenberg-Marquardt algorithm coupled with Monte Carlo modeling for the error bar and a Bayesian statistics method. The Bayesian method computes the probability distribution for the measured photons and electron temperature and this information can be used to ensemble data and will allow future integrated data analysis efforts.

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