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Williams M.M.,Centers for Disease Control and Prevention | Taylor T.H.,Centers for Disease Control and Prevention | Warshauer D.M.,Wisconsin State Laboratory of Hygiene | Martin M.D.,Centers for Disease Control and Prevention | And 2 more authors.
Journal of Clinical Microbiology | Year: 2015

Real-time PCR (rt-PCR) is an important diagnostic tool for the identification of Bordetella pertussis, Bordetella holmesii, and Bordetella parapertussis. Most U.S. public health laboratories (USPHLs) target IS481, present in 218 to 238 copies in the B. pertussis genome and 32 to 65 copies in B. holmesii. The CDC developed a multitarget PCR assay to differentiate B. pertussis, B. holmesii, and B. parapertussis and provided protocols and training to 19 USPHLs. The 2012 performance exercise (PE) assessed the capability of USPHLs to detect these three Bordetella species in clinical samples. Laboratories were recruited by the Wisconsin State Proficiency Testing program through the Association of Public Health Laboratories, in partnership with the CDC. Spring and fall PE panels contained 12 samples each of viable Bordetella and non-Bordetella species in saline. Fifty and 53 USPHLs participated in the spring and fall PEs, respectively, using a variety of nucleic acid extraction methods, PCR platforms, and assays. Ninety-six percent and 94% of laboratories targeted IS481 in spring and fall, respectively, in either singleplex or multiplex assays. In spring and fall, respectively, 72% and 79% of USPHLs differentiated B. pertussis and B. holmesii and 68% and 72% identified B. parapertussis. IS481 cycle threshold (CT) values for B. pertussis samples had coefficients of variation (CV) ranging from 10% to 28%. Of the USPHLs that differentiated B. pertussis and B. holmesii, sensitivity was 96% and specificity was 95% for the combined panels. The 2012 PE demonstrated increased harmonization of rt-PCR Bordetella diagnostic protocols in USPHLs compared to that of the previous survey. Copyright © 2015, American Society for Microbiology. All Rights Reserved.


Park S.-S.,Chonnam National University | Park S.-S.,University of Wisconsin - Madison | Schauer J.J.,University of Wisconsin - Madison | Schauer J.J.,Wisconsin State Laboratory of Hygiene | Cho S.-Y.,Chonnam National University
Atmospheric Environment | Year: 2013

24-hPM2.5 samples were collected at a roadway site every 6th day for one year (September 2010 through August 2011) and analyzed for organic carbon (OC), elemental carbon (EC), water-soluble organic carbon (WSOC), hydrophilic and hydrophobic fractions of WSOC (WSOCHPI and WSOCHPO), and ionic species, to provide important seasonal quantitative information on the primary and secondary sources of two WSOC fractions. Five minute black carbon (BC) concentrations were also measured using a seven-channel wavelength aethalometer to investigate the relationship of biomass burning (BB)-derived BC data from a BC@880nm tracer method and WSOC. There has been increased interest in the light adsorption of WSOC and water-insoluble OC but most of the tools that have been used to understand these relationships have limited to extracts of filter-based samples. The impact of BB emissions on WSOC fractions was examined using the relationship between δBC (=BC@370nm-BC@880nm) and WSOC (or K+), and between BB tracers (WSOC and K+) and BB-derived BC (BCBB) estimated. The moderate correlation (R2=0.41) of WSOC and δBC during the cold months of November through April may support the contribution of BB emissions to the observed WSOC. Predicted BCBB correlated well with K+, WSOCHPI, and WSOCHPO concentrations (R2 of 0.65, 0.43, and 0.61, respectively), suggesting BB emissions may have an influence on the WSOC fractions observed. Contributions of non-BB, BB, and secondary OC (SOC) to both WSOCHPI and WSOCHPO were estimated using a multiple linear regression analysis. The monthly average contribution of non-BB emissions ranged from 12.6% to 29.4% of the WSOCHPI and from 21.5% to 44.1% of the WSOCHPO, with high contributions occurring during the cold months and low contributions occurred during the warm months. BB emissions contributed more to WSOCHPI (2.7%-13.1%) than WSOCHPO (0.2%-1.1%), and the SOC contribution to both WSOC fractions was significant. SOC accounted for 57.2%-79.7% of the WSOCHPI, and for 55.3%-78.0% of the WSOCHPO, with the higher fractions occurring in summer. During the summer the high SOC contributions were consistent with high OC/EC and WSOC/OC ratios. Results of this study suggest that non-BB ("fossil") emissions are likely an important source to WSOCHPI and WSOCHPO observed at this roadway site. © 2013 Elsevier Ltd.


Park S.-S.,Chonnam National University | Sim S.Y.,Chonnam National University | Bae M.-S.,Mokpo National University | Schauer J.J.,University of Wisconsin - Madison | Schauer J.J.,Wisconsin State Laboratory of Hygiene
Atmospheric Environment | Year: 2013

Size-resolved measurements of particulate matter (PM) emissions from 10 biomass materials (rice straw, soybean stem, green perilla stem, red pepper stem, pine needles, cherry leaves, cherry stem, maple leaves, gingko leaves and gingko stem) were conducted in a laboratory hood chamber environment using a 10-stage MOUDI. Samples were analyzed to determine the mass, water soluble organic carbon (WSOC), and water soluble inorganic species. This study examines how particle emissions and size distributions of chemical components vary with biomass materials. Mass fractions of water soluble organic mass (WSOM) (=1.6×WSOC) and ionic species to the PM1.8 emissions varied significantly depending on the biomass type burned. The percent mass of WSOM in PM1.8 emissions ranged from 19.8% (green perilla stem) to 41.9% (red pepper stem) for agricultural crop residues, while the tree category accounted for 9.6% (gingko leaves) to 44.0% (gingko stem) of the PM1.8 emissions. Total ionic species contents in the PM1.8 mass ranged from 7.4% (rice straw) to 26.9% (green perilla stem) for the agricultural waste category, and 5.8% (maple leaves) to 23.5% (gingko stem) for the tree category. The ionic species fraction of the PM1.8 emission was dominated by K+, Cl-, and SO4 2-, while Ca2+ was important in the coarse mode particles (>3.1μm). PM1.8 emissions of K+, Cl-, and SO4 2- were as high as 16.9%, 9.0%, and 5.8%, respectively, and were from the green perilla stem, red pepper stem, and gingko stem emissions. Normalized size distributions of mass, WSOC, K+, Cl-, SO4 2-, and oxalate in the biomass burning emissions showed a unimodal size distribution, peaking in the size ranges of 0.32-0.55μm and 0.55-1.0μm. Size-resolved PM mass fractions of WSOM, K+, Cl-, and SO4 2- showed fairly consistent distributions for each biomass type, with higher fractions in the ultrafine mode (<0.10μm) and lower fractions in the accumulation mode of 0.32-1.0μm. The size distributions of WSOC were strongly correlated (mostly R2>0.90) with those of K+ in the particle size range of <0.1μm and 0.1-1.8μm and the biomass types. Strong correlations between the concentrations of K+-Cl- and K+-SO4 2- were observed for the following size ranges; <0.1μm, 0.1-1.0μm, 1.0-1.8μm, and 1.8-3.1μm for most biomass burning emissions. Regression line slopes for K+/WSOC (i.e., mass of K+/mass of WSOC from biomass burning emissions) were not significantly changed for particle size and biomass type, but slopes for Cl-/K+ and SO4 2-/K+ varied significantly with the particle size (ultrafine, condensation, droplet, and coarse modes) and biomass type. © 2013 Elsevier Ltd.


Jones A.W.,National Board of Forensic Medicine | Harding P.,Wisconsin State Laboratory of Hygiene
Forensic Science International | Year: 2013

The aim of this study was to evaluate the characteristics of traffic offenders with unusually high blood-alcohol concentrations (BAC. >. 0.4. g%) when arrested. The BAC that kills one person might be easily tolerated by another, depending on, among other things, the person's age, pattern of drinking, and the development of tolerance. The archives of two forensic laboratories, one in Sweden and the other in Wisconsin (USA), were searched to find traffic offenders with BACs. >. 0.4. g%. The results were compared in relation to the person's age and gender, mean BAC and the weekday and time of day of the arrest. The mean age (±standard deviation) of N= 158 Swedish offenders was 45. ±. 9.0. y, which was not significantly different from the 43. ±. 9.4. y in N= 233 Wisconsin drivers (p>. 0.05). Overall there were more men (78%) than women (22%) arrested with BAC's. >. 0.4. g%, although the proportion of women in Wisconsin (35%) was higher than in Sweden (9%) (p<. 0.001). The mean (median) and highest BAC did not differ between jurisdictions; 0.429. g% (0.422) and 0.546. g% in Sweden and 0.428. g% (0.421. g%) and 0.526. g% in Wisconsin. In Sweden 40% of the arrests occurred on Fridays and Saturdays, whereas in Wisconsin the arrests of people with such high BAC's were more evenly distributed throughout the week. Forty eight percent of the arrests in Sweden were made between 12 noon and 6. pm compared with 37% in Wisconsin. Neither the mean age of offenders nor their mean BAC seemed to depend on the weekday or time of day of the arrest. Attempting to drive with a BAC above 0.4. g% verifies the development of an appreciable tolerance to ethanol-induced cognitive and psychomotor impairment. Reaching such a high BAC probably requires continuous heavy drinking over several days as opposed to an evening's binge drinking. © 2013 Elsevier Ireland Ltd.


Wells J.,University of Wisconsin - Madison | Rosenberg M.,University of Wisconsin - Madison | Hoffman G.,Wisconsin State Laboratory of Hygiene | Anstead M.,University of Kentucky | Farrell P.M.,University of Wisconsin - Madison
Pediatrics | Year: 2012

OBJECTIVES: Because cystic fibrosis can be difficult to diagnose and treat early, newborn screening programs have rapidly developed nationwide but methods vary widely. We therefore investigated the costs and consequences or specific outcomes of the 2 most commonly used methods. METHODS: With available data on screening and follow-up, we used a simulation approach with decision trees to compare immunoreactive trypsinogen (IRT) screening followed by a second IRT test against an IRT/DNA analysis. By using a Monte Carlo simulation program, variation in the model parameters for counts at various nodes of the decision trees, as well as for costs, are included and applied to fictional cohorts of 100 000 newborns. The outcome measures included the numbers of newborns given a diagnosis of cystic fibrosis and costs of screening strategy at each branch and cost per newborn. RESULTS: Simulations revealed a substantial number of potential missed diagnoses for the IRT/IRT system versus IRT/DNA. Although the IRT/IRT strategy with commonly used cutoff values offers an average overall cost savings of $2.30 per newborn, a breakdown of costs by societal segments demonstrated higher out-of-pocket costs for families. Two potential system failures causing delayed diagnoses were identified relating to the screening protocols and the follow-up system. CONCLUSIONS: The IRT/IRT screening algorithm reduces the costs to laboratories and insurance companies but has more system failures. IRT/DNA offers other advantages, including fewer delayed diagnoses and lower out-of-pocket costs to families. Copyright © 2012 by the American Academy of Pediatrics.


Corsi S.R.,U.S. Geological Survey | Graczyk D.J.,U.S. Geological Survey | Geis S.W.,Wisconsin State Laboratory of Hygiene | Booth N.L.,U.S. Geological Survey | Richards K.D.,U.S. Geological Survey
Environmental Science and Technology | Year: 2010

A new perspective on the severity of aquatic toxicity impact of road salt was gained by a focused research effort directed at winter runoff periods. Dramatic impacts were observed on local, regional, and national scales. Locally, samples from 7 of 13 Milwaukee, Wisconsin area streams exhibited toxicity in Ceriodaphnia dubia and Pimephales promelas bioassays during road-salt runoff. Another Milwaukee stream was sampled from 1996 to 2008 with 72% of 37 samples exhibiting toxicity in chronic bioassays and 43% in acute bioassays. The maximum chloride concentration was 7730 mg/L. Regionally, in southeast Wisconsin, continuous specific conductance was monitored as a chloride surrogate in 11 watersheds with urban land use from 6.0 to 100%. Elevated specific conductance was observed between November and April at all sites, with continuing effects between May and October at sites with the highest specific conductance. Specific conductance was measured as high as 30 800 μS/cm (Cl = 11 200 mg/L). Chloride concentrations exceeded U.S. Environmental Protection Agency (USEPA) acute (860 mg/L) and chronic (230 mg/L) water-quality criteria at 55 and 100% of monitored sites, respectively. Nationally, U.S. Geological Survey historical data were examined for 13 northern and 4 southern metropolitan areas. Chloride concentrations exceeded USEPA water-quality criteria at 55% (chronic) and 25% (acute) of the 168 monitoring locations in northern metropolitan areas from November to April. Only 16% (chronic) and 1% (acute) of sites exceeded criteria from May to October. At southern sites, very few samples exceeded chronic water-quality criteria, and no samples exceeded acute criteria. © 2010 American Chemical Society.


Stoiber T.L.,University of Wisconsin - Madison | Shafer M.M.,University of Wisconsin - Madison | Shafer M.M.,Wisconsin State Laboratory of Hygiene | Armstrong D.E.,University of Wisconsin - Madison
Environmental Toxicology | Year: 2013

The toxicity of metals to organisms is, in-part, related to the formation of reactive oxygen species (ROS) in cells and subsequent oxidative stress. ROS are by-products of normal respiration and photosynthesis processes in organisms, but environmental factors, like metal exposure, can stimulate excess production. Metals involved in several different mechanisms such as Haber-Weiss cycling and Fenton-type reactions can produce ROS. Some metals, such as Cd, may contribute to oxidative stress indirectly by depleting cellular antioxidants. We investigated the measurement of ROS as a sensitive biomarker of metal toxicity (that could possibly be implemented in a biotic ligand model for algae) and we compared ROS induction in response to several contrasting transition metals (Cu, V, Ni, Zn, and Cd). We also compared the ROS response to glutathione and growth toxicity endpoints measured in a previous study. The cell-permeable dye, 2′7′dichlorodihydrofluorescein diacetate, was used as a probe to detect formation of ROS in Chlamydomonas reinhardtii cells. Metal-exposed cells were incubated with the fluorescent dye in a 96-well plate and monitored over 5.5 h. A dose-response of ROS formation was observed with Cu exposure in the range of 20-500 nM. Cu produced more ROS compared with either Zn or Cd (both nonredox active metals). The redox-active metal V produced increased ROS with increased concentration. The measurement of ROS may be a useful indicator of Cu toxicity, but the signal to noise ratio was better for the glutathione endpoint assay. © 2011 Wiley Periodicals, Inc.


Stoiber T.L.,University of Wisconsin - Madison | Shafer M.M.,University of Wisconsin - Madison | Shafer M.M.,Wisconsin State Laboratory of Hygiene | Armstrong D.E.,University of Wisconsin - Madison
Environmental Toxicology and Chemistry | Year: 2010

The toxicity of cadmium to aquatic organisms is well known, but the mechanisms of toxicity are not as clearly understood. In the present study, Cd bioassay experiments incorporating both traditional endpoints and novel thiol-based endpoints were conducted with Chlamydomonas reinhardtii. The results were compared with results from previous bioassay experiments to probe the apparent contrasting biochemical mechanisms of toxicity of copper and cadmium as expressed in cellular glutathione and the glutathione cycle. Total glutathione and reduced to oxidized glutathione ratio (GSH/GSSG) measurements were remarkably different in Cd-compared with Cu-exposed cells. Whereas total glutathione in cells decreased with increasing Cu concentration, Cd caused dramatic increases. Total glutathione increased by 4.5-fold with 80 nM Cd treatment over concentrations in Cd-free controls. Glutathione reductase (GR) enzyme activity was positively correlated (r2Cu = 0.96, r 2Cd = 0.85) with glutathione concentrations for both metals. Measurements of mRNA for GR were increased 2-fold in response to Cd exposure (80 nM) and correlated well with GR enzyme activity. Glutathione concentrations and GR enzyme activity are useful endpoints for both Cu and Cd toxicity in algae, even though the metals elicit opposing responses. We conclude that Cu decreases glutathione concentrations by inhibiting GR enzyme activity. In contrast, Cd stimulates GR enzyme activity and increases glutathione concentrations as cells respond to Cd-induced stress by producing increased antioxidant capacity. The present study demonstrates that determining the glutathione response in cells is important for understanding the metalspecific mechanisms of toxicity and that these associated novel endpoints may be useful metrics for accurately predicting toxicity. © 2009 SETAC.


Stoiber T.L.,University of Wisconsin - Madison | Shafer M.M.,Wisconsin State Laboratory of Hygiene | Armstrong D.E.,University of Wisconsin - Madison
Environmental Toxicology and Chemistry | Year: 2012

In the present study, the adsorption and uptake of copper (Cu) and cadmium (Cd) in Chlamydomonas reinhardtii were examined to establish fundamental toxicity relationships to glutathione and cell-growth endpoints. Establishing these fundamental relationships of metal accumulation and toxicity metrics is necessary to subsequently implement an algal biotic ligand model. The glutathione response was similar to the response measured from growth endpoints for both internal and adsorbed Cu, indicating that glutathione may be a useful biomarker of toxicity. The glutathione response with Cd contrasted markedly with that observed with Cu and was therefore observed to be a metal-specific biomarker. The density of sites binding metals and the related stability constants for the algal cell surface were also determined. Short exposures to metals (2h) were conducted, and we determined 6.0×10 -6mol/g sites binding Cu and 2.0×10 -6mol/g sites binding Cd and conditional stability constants as log K′=7.2 and log K′=6.7 for Cu and Cd, respectively. Experiments were also conducted to determine the effect on toxicity endpoints of varying nitrate concentrations and different humic acids (HA) in the exposure media. Varying nitrate concentrations did not have an effect on cell growth over 24h. The surface-adsorbed Cu measurements from the experiments with HA depended on the type and concentration of HA. © 2011 SETAC.


Dequattro Z.A.,University of Wisconsin - Madison | Peissig E.J.,University of Wisconsin - Madison | Antkiewicz D.S.,Wisconsin State Laboratory of Hygiene | Lundgren E.J.,University of Wisconsin - Madison | And 3 more authors.
Environmental Toxicology and Chemistry | Year: 2012

High concentrations (375ng/L) of the steroid hormone progesterone (P4) were measured in snowmelt runoff associated with large livestock-feeding operations in Wisconsin. To gain insight into the potential endocrine-disrupting effects of P4 in fish, experiments were conducted to evaluate the effects of short-term exposure to environmentally relevant concentrations of P4 on reproduction and embryonic development in the fathead minnow (Pimephales promelas). For the reproduction assay, groups of reproductively mature fish were exposed for 21d to nominal concentrations of 0, 10, 100, and 1,000ng/L P4 in a flow-through system, and various key reproductive endpoints (e.g., egg number, fertilization success) were quantified throughout the exposure period. The embryonic development assay consisted of incubating fathead minnow eggs in static culture to quantify the effects of P4 on early development and hatching success. Progesterone caused dose-dependent decreases in fecundity and fertility and significantly reduced gonadosomatic index and vitellogenin gene expression in females. There were no effects of P4 on early embryonic development or hatching success. Progesterone may be a significant endocrine-disrupting chemical in fish. © 2012 SETAC.

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