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Abstract Background We sought to determine if cold temperature and snowfall are independently associated with stroke mortality, and whether effects differ between hemorrhagic and ischemic stroke. Materials and methods We conducted a case-crossover study of 13,201 stroke deaths utilizing weather records between the months of November and April for Quebec, Canada from 1981 to 2015. We compared exposure to cold temperature and snowfall with controls days when stroke death did not occur. We computed odds ratios (OR) and 95% confidence intervals (CI) for the association of minimum temperature and duration of snowfall with stroke, adjusted for change in barometric pressure and relative humidity. Results The likelihood of mortality the day following exposure to cold temperature was elevated for hemorrhagic stroke in men, independent of snowfall. Relative to 0 °C, a temperature of −20 °C was associated with 1.17 times the odds of hemorrhagic stroke death (95% CI 1.04–1.32). An independent effect of snowfall was also present in men, with 12 h of snowfall associated with 1.12 times the odds of hemorrhagic stroke death (95% CI 1.00–1.24) compared with no snowfall. There was no evidence of an increased risk in women. Cold temperature and snowfall were not associated with ischemic stroke death in either men or women. Conclusion Our results suggest that cold temperature and snowfall are independent risk factors for death from hemorrhagic stroke in men. These findings imply that interventions to prevent fatal hemorrhagic stroke during winter should include both cold temperature exposure and snowfall in men. Highlights • We studied the independent effects of cold temperature and snowfall on stroke. • We analyzed 13,201 stroke deaths using a case-crossover study design. • Cold temperature and snowfall were associated with hemorrhagic stroke in men. • Cold temperature and snowfall were not associated with ischemic stroke. • There was no association with the risk of stroke in women. [ABSTRACT FROM AUTHOR]

People experiencing homelessness have an increased risk of cold-related injuries. This study determined the rate of emergency department (ED) visits for cold-related injuries among homeless adults and low-income controls in Toronto, Canada. Homeless individuals were recruited at shelters and meal programs. Age- and sex-matched controls living in low-income neighborhoods were selected. ED utilization was ascertained over 4-years of follow-up (2005-9) using administrative databases. A total of 16 ED visits for cold-related injuries were observed among 587 homeless men and 296 homeless women. The rate of ED visits was 6.7 (95% CI, 4.2-12.4) per 1000 person-years of observation among homeless men and 0.9 (95% CI, 0.03-5.6) among homeless women. ED visit rates were significantly higher among homeless men compared to low-income men (P < 0.001) and significantly higher among homeless men compared to homeless women (P = 0.03). Targeted public health interventions are needed to reduce the risk of cold-related injuries among people experiencing homelessness. [ABSTRACT FROM AUTHOR]

Due to its high ductility, weldability and toughness at low temperature, CSA G40.21-350WT steel in Canada is primarily used in bridge construction, ship building, and seismic energy dissipation systems. This article presents uniaxial tensile tests and constant- and variable-amplitude cyclic testing performed on 350WT steel at room and subfreezing temperatures. The variable-amplitude tests include common step-loading patterns as well as tests under strain signals obtained from the brace response in building structures subjected to three different types of earthquakes. The ductility of 350WT steel from monotonic tensile tests is essentially same at room and low temperatures (−40 °C). The cyclic test results revealed that cold temperatures as low as −35 °C did not have adverse effects on the low cycle fatigue life of 350WT steel. The benchmark constant-amplitude tests were employed to predict fatigue life under different large-strain variable-amplitude loading patterns both for room and subfreezing temperature conditions. In addition to the common strain-life approach, the adequacy of two well-established energy-life models for predicting fatigue life under variable-amplitude loading was evaluated. The strain-life approach generally performed better than the energy-life methods, particularly for step-loading histories. Comparison between predictions and laboratory observations showed that the fatigue failure life under large strain seismic loading can be accurately estimated, especially at room temperature. [ABSTRACT FROM AUTHOR]

Background: West Nile virus (WNV) is the leading cause of mosquito-borne disease in humans in the United States. Since the introduction of the disease in 1999, incidence levels have stabilized in many regions, allowing for analysis of climate conditions that shape the spatial structure of disease incidence., Objectives: Our goal was to identify the seasonal climate variables that influence the spatial extent and magnitude of WNV incidence in humans., Methods: We developed a predictive model of contemporary mean annual WNV incidence using U.S. county-level case reports from 2005 to 2019 and seasonally averaged climate variables. We used a random forest model that had an out-of-sample model performance of R 2 = 0.61 ., Results: Our model accurately captured the V-shaped area of higher WNV incidence that extends from states on the Canadian border south through the middle of the Great Plains. It also captured a region of moderate WNV incidence in the southern Mississippi Valley. The highest levels of WNV incidence were in regions with dry and cold winters and wet and mild summers. The random forest model classified counties with average winter precipitation levels < 23.3 mm / month as having incidence levels over 11 times greater than those of counties that are wetter. Among the climate predictors, winter precipitation, fall precipitation, and winter temperature were the three most important predictive variables., Discussion: We consider which aspects of the WNV transmission cycle climate conditions may benefit the most and argued that dry and cold winters are climate conditions optimal for the mosquito species key to amplifying WNV transmission. Our statistical model may be useful in projecting shifts in WNV risk in response to climate change. https://doi.org/10.1289/EHP10986.

A mobile testing center was installed at a lagoon wastewater treatment plant (WWTP) at Terrebonne, Canada to investigate the rate of ammonia removal of attached growth treatment systems at 4°C and at low influent carbon concentrations. The testing center housed two laboratory-scale reactors, a pilot-scale BioStyr system (Veolia Water) and a pilot-scale moving bed bioreactor (MBBR) system (Veolia Water). Although the rates of laboratory-scale and the pilot-scale systems demonstrated that the exposure time to low temperature has a significant effect on the kinetics of the system, the ammonia removal rates of all the systems were shown to be significant at 4°C. A strong correlation was demonstrated between the rates of ammonia removal produced by the laboratory-scale reactors, the pilot BioStyr system and pilot MBBR system; thus verifying the scale-up capability of the laboratory-scale reactors and demonstrating that nitrifiers can achieve ammonia removal under cold temperature conditions for elapsed periods of time independent of the reactor design. Finally, the ammonia removal rates of the laboratory-scale systems, the BioStyr pilot system, and the MBBR pilot system were all accurately predicted by a recently proposed Theta model. [ABSTRACT FROM AUTHOR]

Insects that hatch in winter and early spring in desert and montane regions are likely to encounter extreme weather events, including precipitous drops in temperature. The susceptibility of insects to exposure to subzero temperatures is predicted to decrease with increasing latitude or elevation. Mormon crickets occur over a broad latitudinal range from southwestern United States to Canada and a broad elevational range from near sea-level to 3,000 m. Population declines have been attributed to late freezing events, but winter hatching suggests they may also be cold tolerant. Lower lethal temperature of high elevation populations in low latitude Arizona (AZ) and high latitude Wyoming (WY) was measured by exposing nymphs and adults to 6 h or 24 h of subzero temperature. From similar latitude, WY was compared with mid-elevation Idaho (ID) and low elevation Oregon (OR) populations. Contrary to the prediction, lethal temperature of third instar nymphs was lower in AZ than in the more northerly populations. Consistently, AZ was more tolerant of cold in early nymphal instars relative to populations from higher latitude. Early hatching at lower latitudes might increase the risk of early instars experiencing a severe cold snap relative to nymphs at high latitudes. Also, contrary to prediction, the lethal temperature of adults increased with elevation, whereas third instar nymphs from mid-elevation ID were the most susceptible to cold exposure. Cold tolerance in immature and mature stages is more likely to be uncoupled when life stages do not coincide, as with Mormon crickets., (Published by Oxford University Press on behalf of Entomological Society of America 2021.)

Higher transmissibility of SARS-CoV-2 in cold and dry weather conditions has been hypothesized since the onset of the COVID-19 pandemic but the level of epidemiological evidence remains low. During the first wave of the pandemic, Spain, Italy, France, Portugal, Canada and USA presented an early spread, a heavy COVID-19 burden, and low initial public health response until lockdowns. In a context when testing was limited, we calculated the basic reproduction number (R 0 ) in 63 regions from the growth in regional death counts. After adjusting for population density, early spread of the epidemic, and age structure, temperature and humidity were negatively associated with SARS-CoV-2 transmissibility. A reduction of mean absolute humidity by 1 g/m 3 was associated with a 0.15-unit increase of R 0 . Below 10 °C, a temperature reduction of 1 °C was associated with a 0.16-unit increase of R 0 . Our results confirm a dependency of SARS-CoV-2 transmissibility to weather conditions in the absence of control measures during the first wave. The transition from summer to winter, corresponding to drop in temperature associated with an overall decrease in absolute humidity, likely contributed to the intensification of the second wave in north-west hemisphere countries. Non-pharmaceutical interventions must be adjusted to account for increased transmissibility in winter conditions.

Background: Platelets are the most commonly discarded blood product in Canada, with the most common cause of in-date product loss being improper storage. Transport containers to maintain temperature and extend acceptable return time may represent a method to reduce wastage. The objective of this study was to evaluate the impact of a validated Platelet Transport Bag (PTB) on platelet wastage., Study Design and Methods: Thirty-six hospitals with the highest platelet discards were invited to participate in a before-after observational study. Hospitals were instructed to utilize a validated 4-h PTB for clinical situations where immediate transfusion was not planned. Five hospitals audited in-date platelet discards from July 2018 to November 2019 to characterize wastage causes. In-date platelet discard data 12 months before and after the start date for each site were analyzed to determine changes in wastage., Results: Of 36 hospital sites, 16 agreed to participate. Pre- and postdiscards were 277 and 301, respectively, for all sites combined. There were no significant before-after change in wastage rate (+0.05%, p = .51). Fifty discards were included in the detailed audit; the most common reasons were return to the blood bank after more than 60 min outside a PTB (n = 17, 34%) and return in a red cell cooler (n = 10, 20%)., Conclusion: Implementation of PTB did not improve wastage. Common causes of in-date discards were return after 1 h outside of a PTB and placement in a red cell cooler in error. Further research is required to investigate potential strategies to mitigate in-date platelet wastage., (© 2021 AABB.)

Context: A number of studies have shown that cold has an important impact on human health. However, almost no studies focused on cold warning systems to prevent those health effects. For Nordic regions, like the province of Quebec in Canada, winter is long and usually very cold with an observed increase in mortality and hospitalizations throughout the season. However, there is no existing system specifically designed to follow in real-time this mortality increase throughout the season and to alert public health authorities prior to cold waves., Objective: The aim is to establish a watch and warning system specifically for health impacts of cold, applied to different climatic regions of the province of Quebec., Methodology: A methodology previously used to establish the health-heat warning system in Quebec is adapted to cold. The approach identifies cold weather indicators and establishes thresholds related to extreme over-mortality or over-hospitalization events in the province of Quebec, Canada., Results and Conclusion: The final health-related thresholds proposed are between (-15 °C, -23 °C) and (-20 °C, -29 °C) according to the climatic region for excesses of mortality, and between (-13 °C, -23 °C) and (-17 °C, -30 °C) for excesses of hospitalization. These results suggest that the system model has a high sensitivity and an acceptable number of false alarms. This could lead to the establishment of a cold-health watch and warning system with valid indicators and thresholds for each climatic region of Quebec. It can be seen as a complementary system to the existing one for heat warnings, in order to help the public health authorities to be well prepared during an extreme cold event., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Aerated lagoons, typically used by small communities, often provide limited removal of wastewater nutrients. Given increasingly stringent wastewater standards, it is imperative that effective, but economical and easy-to-operate, treatment technologies be developed. The Submerged Attached Growth Reactor (SAGR®) is a treatment process developed to perform nitrification near freezing temperatures. Previous tests on full-scale installations have shown that SAGR could consistently remove ammonia to below current Canadian standards and provide additional total suspended solids and biochemical oxygen demand removal. In this study, we evaluated removal of polar chemicals of emerging concern (CECs), including pharmaceuticals, personal care products, and pesticides, at SAGR installations in two Manitoba First Nations communities (MCN and LPFN) under cold winter conditions. Both showed some removal of diclofenac, naproxen, clarithromycin, metoprolol, and trimethoprim, likely by biotransformation. Average naproxen removal was 21% (2.53 × 10 3  ng L -1 ) in MCN and 64% (1.58 × 10 3  ng L -1 ) in LPFN. Atenolol was well-removed by SAGR, by 80% on average (range of 64%-94%). Clarithromycin, metoprolol, and trimethoprim removal was similar within and between systems, ranging from 54% to 76% (30.8-3.07 × 10 2  ng L -1 removed). Carbamazepine was detected in nearly all samples, but was not well-removed, consistent with other treatment studies. Overall, results showed that SAGR technology could moderately remove CECs, while providing the designed treatment performance for other parameters. This work will help to improve our understanding of wastewater treatment in small and/or remote communities with limited infrastructure and challenging cold-weather conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Low temperatures limit the distribution and abundance of ectotherms. However, many insects can survive low temperatures by employing one of two cold tolerance strategies: freeze avoidance or freeze tolerance. Very few species can employ both strategies, but those that do provide a rare opportunity to study the mechanisms that differentiate freeze tolerance and freeze avoidance. We showed that overwintering pupae of the cabbage white butterfly Pieris rapae can be freeze tolerant or freeze avoidant. Pupae from a population of P. rapae in northeastern Russia (Yakutsk) froze at c. -9.3 °C and were freeze-tolerant in 2002-2003 when overwintered outside. However, P. rapae from both Yakutsk and southern Canada (London) acclimated to milder laboratory conditions in 2014 and 2017 froze at lower temperatures (< -20 °C) and were freeze-avoidant. Summer-collected P. rapae larvae (collected in Yakutsk in 2016) were partially freeze-tolerant, and decreased the temperature at which they froze in response to starvation at mild low temperatures (4 °C) and repeated partial freezing events. By comparing similarly-acclimated P. rapae pupae from both populations, we identified molecules that may facilitate low temperature tolerance, including the hemolymph ice-binding molecules and several potential low molecular weight cryoprotectants. Pieris rapae from Yakutsk exhibited high physiological plasticity, accumulating cryoprotectants and almost doubling their hemolymph osmolality when supercooled to -15 °C for two weeks, while the London P. rapae population exhibited minimal plasticity. We hypothesize that physiological plasticity is an important adaptation to extreme low temperatures (i.e. in Yakutsk) and may facilitate the transition between freeze avoidance and freeze tolerance., Competing Interests: Declaration of Competing Interest None. The authors have no competing interests to declare., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Winter is an understudied but key period for the socioecological systems of northeastern North American forests. A growing awareness of the importance of the winter season to forest ecosystems and surrounding communities has inspired several decades of research, both across the northern forest and at other mid- and high-latitude ecosystems around the globe. Despite these efforts, we lack a synthetic understanding of how winter climate change may impact hydrological and biogeochemical processes and the social and economic activities they support. Here, we take advantage of 100 years of meteorological observations across the northern forest region of the northeastern United States and eastern Canada to develop a suite of indicators that enable a cross-cutting understanding of (1) how winter temperatures and snow cover have been changing and (2) how these shifts may impact both ecosystems and surrounding human communities. We show that cold and snow covered conditions have generally decreased over the past 100 years. These trends suggest positive outcomes for tree health as related to reduced fine root mortality and nutrient loss associated with winter frost but negative outcomes as related to the northward advancement and proliferation of forest insect pests. In addition to effects on vegetation, reductions in cold temperatures and snow cover are likely to have negative impacts on the ecology of the northern forest through impacts on water, soils, and wildlife. The overall loss of coldness and snow cover may also have negative consequences for logging and forest products, vector-borne diseases, and human health, recreation, and tourism, and cultural practices, which together represent important social and economic dimensions for the northern forest region. These findings advance our understanding of how our changing winters may transform the socioecological system of a region that has been defined by the contrasting rhythm of the seasons. Our research also identifies a trajectory of change that informs our expectations for the future as the climate continues to warm., (© 2019 The Authors. Ecological Applications published by Wiley Periodicals, Inc. on behalf of Ecological Society of America.)

Background: Dietary Reference Intakes are used to guide the energy intake of the Canadian Armed Forces (CAF) field rations provided to military personnel deployed for training or operations. However, the high energy expenditures likely to occur under harsh environmental/metabolically challenging deployment conditions may not be adequately considered. This study examined the Ad libitum energy and nutrient intakes of CAF personnel (n = 18) consuming field rations in a resting thermoneutral environment and during a day of standardized strenuous infantry activities at varying environmental temperatures., Methods: Dietary intake was assessed using a measured food intake/food waste method during the experimental treatment and for 6 h after treatment. Four treatments were administered in a randomized counterbalanced design: exercise (as standardized infantry activities) in the heat (30 °C), exercise in the cold (- 10 °C), exercise in temperate thermoneutral (21 °C) air temperatures and a resting (sedentary) trial (21 °C)., Results: The average Ad libitum consumption of field rations was 70% of the provided total energy (2776 ± 99 kcal/8 h) during all treatments. Even with an acute challenge of increased energy expenditure and temperature stress in the simulated field conditions, participants' energy intakes (1985 ± 747 kcal/8 h) under hot, cold and temperate treatments did not differ from energy intake during the sedentary condition (1920 ± 640 kcal/8 h). Participants' energy intakes (1009 ± 527 kcal/6 h) did not increase during the 6 h posttreatment period when the stresses of the strenuous physical activities and the harsh environmental temperatures had subsided., Conclusion: These results should be considered when planning the provision of field rations for CAF personnel expected to be engaged in strenuous physical activities with prolonged exposure to temperature extremes.

As a result of anthropization and industrialization, northern remote communities face issues of soil contamination by mixtures of organic and inorganic contaminants. Soil bioremediation in cold environments is particularly challenging because of slower degradation rates, slower production of biomass for phytoextraction of trace elements (TEs), and remoteness, which can complicate logistics and inflate costs. This study evaluated a decontamination approach integrating indigenous willows, fungi and compost in a northern community. The site was a waste oil pit and its soil was initially contaminated with petroleum hydrocarbons (PHC) exceeding 200 g kg -1 and TEs including As, Cd, Co, Cr, Cu, Pb and Zn. In under five years, 65 and 75% of PHC (C6-C50 and >C50) were degraded, compared to 27 and 13% for the untreated control soil. We found contrasting TE translocation patterns to the aboveground biomass for the willow species used (Salix planifolia and Salix alaxensis), as well as distinctive rooting strategies. Hazard quotients were calculated to assess the risk plant material could pose to local wildlife. The highest TE concentration measured was Zn in S. planifolia, which exceeded Canadian soil guidelines. Results indicate toxicity risks to animals linked to TEs in Salix spp. leaves is generally unlikely. The fungus Trametes versicolor inoculated into the soil did not fruit, however fruiting bodies of Psathyrella sp. were observed consistently (four out of five years). Biological tests indicated that in five growing seasons soil toxicity significantly decreased compared to the untreated soil used as control. This was demonstrated by vegetation cover (137 vs 11% cover), toxicity assays on earthworms (Eisenia andrei) (0 vs 33% mortality) and barley seed germination (Hordeum vulgare) (86 vs 62% germination). The proposed decontamination approach, without the use of synthetic fertilizers, is promising for the PHC remediation of mixed-contaminants on cold climate sites., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Treeline advance has occurred throughout the twentieth century in mountainous regions around the world; however, local variation and temporal lags in responses to climate warming indicate that the upper limits of some treelines are not necessarily in climatic equilibrium. These observations suggest that factors other than climate are constraining tree establishment beyond existing treelines. Using a seed addition experiment, we tested the effects of seed availability, predation and microsite limitation on the establishment of two subalpine tree species (Picea engelmannii and Abies lasiocarpa) across four treelines in the Canadian Rocky Mountains. The effect of vegetation removal on seedling growth was also determined, and microclimate conditions were monitored. Establishment limitations observed in the field were placed in context with the effects of soil properties observed in a parallel experiment. The seed addition experiment revealed reduced establishment with increasing elevation, suggesting that although establishment within the treeline ecotone is at least partially seed limited, other constraints are more important beyond the current treeline. The effects of herbivory and microsite availability significantly reduced seedling establishment but were less influential beyond the treeline. Microclimate monitoring revealed that establishment was negatively related to growing season temperatures and positively related to the duration of winter snow cover, counter to the conventional expectation that establishment is limited by low temperatures. Overall, it appears that seedling establishment beyond treeline is predominantly constrained by a combination of high soil surface temperatures during the growing season, reduced winter snowpack and unfavourable soil properties. Our study supports the assertion that seedling establishment in alpine treeline ecotones is simultaneously limited by various climatic and nonclimatic drivers. Together, these factors may limit future treeline advance in the Canadian Rocky Mountains and should be considered when assessing the potential for treeline advance in alpine systems elsewhere., (© 2018 John Wiley & Sons Ltd.)

The aim of this study was to determine the course of effects on the microflora on beef carcasses of a commercial dry chilling process in which carcasses were dry chilled for 3 days. Groups of 25 carcasses selected at random were sampled when the chilling process commenced and after the carcasses were chilled for 1, 2, 4, 6, 8, 24, and 67 h for determination of the numbers of aerobes, coliforms, and Escherichia coli. The temperatures of the surfaces and the thickest part of the hip (deep leg) of carcasses, as well as the ambient air conditions, including air temperature, velocity, and relative humidity (RH), were monitored throughout the chilling process. The chiller was operated at 0°C with an off-coil RH of 88%. The air velocity was 1.65 m/s when the chiller was loaded. The initial RH levels of the air in the vicinity of carcasses varied with the locations of carcasses in the chiller and decreased rapidly during the first hour of chilling. The average times for shoulder surfaces, rump surfaces, and the deep leg of carcasses to reach 7°C were 13.6 ± 3.1, 16.0 ± 2.4 and 32.4 ± 3.2 h, respectively. The numbers of aerobes, coliforms, and E. coli on carcasses before chilling were 5.33 ± 0.42, 1.95 ± 0.77, 1.42 ± 0.78 log CFU/4,000 cm(2), respectively. The number of aerobes on carcasses was reduced by 1 log unit each in the first hour of chilling and in the subsequent 23 h of chilling. There was no significant difference (P > 0.05) between the numbers of aerobes recovered from carcasses after 24 and 67 h of chilling. The total numbers (log CFU/100,000 cm(2)) on carcasses before chilling and after the first hour of chilling were 3.86 and 2.24 for coliforms and 3.30 and 2.04 for E. coli. The subsequent 23 h of chilling reduced the numbers of both groups of organisms by a further log unit. No coliforms or E. coli were recovered after 67 h of chilling. The findings show that the chilling regime investigated in this study resulted in significant reductions of all three groups of indicator organisms.

Objectives: To test the hypothesis that challenges to community participation posed by winter weather are greater for individuals who use scooters, manual and power wheelchairs (wheeled mobility devices [WMDs]) than for the general ambulatory population, and to determine what WMD users identify as the most salient environmental barriers to community participation during the winter., Design: Cross-sectional survey organized around 5 environmental domains: technological, natural, physical, social/attitudinal, and policy., Setting: Urban community in Canada., Participants: Convenience sample of WMD users or their proxy (N=99)., Interventions: Not applicable., Main Outcome Measures: Not applicable., Results: Forty-two percent identified reduced outing frequency in winter months, associated with increased age (χ(3)=6.4, P=.04), lack of access to family/friends for transportation (χ(2)=8.1, P=.04), and primary type of WMD used in the winter (scooter χ(2)=8.8, P=.003). Most reported tires/casters becoming stuck in the snow (95%) or slipping on the ice (91%), difficulty ascending inclines/ramps (92%), and cold hands while using controls or pushing rims (85%); fewer identified frozen wheelchair/scooter batteries, seat cushions/backrests, or electronics. Sidewalks/roads were reported to be problematic by 99%. Eighty percent reported needing additional help in the winter. Limited community access in winter led to a sense of loneliness/isolation, and fear/anxiety related to safety. Respondents identified policies that limited participation during winter., Conclusions: People who use WMDs decrease their community participation in cold weather because of multiple environmental barriers. Clinicians, researchers, and policymakers can take a multidimensional approach to mitigate these barriers in order to enhance community participation by WMD users in winter., (Copyright © 2015 American Congress of Rehabilitation Medicine. Published by Elsevier Inc. All rights reserved.)

Pupae of the blueberry maggot, Rhagoletis mendax Curran (Diptera: Tephritidae), were subjected to -20 degrees C for 2, 4, 7, 10, and 15 d (control = 0 d at 4 degrees C) in fall 2011 and 2012 and were allowed to overwinter for at least 181 d at 4 degrees C. Mean adult emergence in the control was 64% in 2012 and 39% in 2013. No adults emerged in May 2012 from 500 pupae treated for > 2 d at -20 degrees C in fall 2011. Two adults emerged in May 2013 from 100 pupae treated for 2 d at -20 degrees C in fall 2012. No adults emerged in May 2013 from 400 pupae subjected to -20 degrees C for durations of 4, 7, 10, and 15 d. No adults emerged from larvae concealed in berries subjected to -20 degrees C for > 2 d. Exposure of larvae and pupae to cold temperatures (i.e., -20 degrees C) for > 2 d can be a valuable nonpesticidal method for killing R. mendax larvae and pupae associated with reusable containers.

Permafrost-affected soils are among the most obvious ecosystems in which current microbial controls on organic matter decomposition are changing as a result of global warming. Warmer conditions in polygonal tundra will lead to a deepening of the seasonal active layer, provoking changes in microbial processes and possibly resulting in exacerbated carbon degradation under increasing anoxic conditions. To identify current microbial assemblages in carbon rich, water saturated permafrost environments, four polygonal tundra sites were investigated on Herschel Island and the Yukon Coast, Western Canadian Arctic. Ion Torrent sequencing of bacterial and archaeal 16S rRNA amplicons revealed the presence of all major microbial soil groups and indicated a local, vertical heterogeneity of the polygonal tundra soil community with increasing depth. Microbial diversity was found to be highest in the surface layers, decreasing towards the permafrost table. Quantitative PCR analysis of functional genes involved in carbon and nitrogen-cycling revealed a high functional potential in the surface layers, decreasing with increasing active layer depth. We observed that soil properties driving microbial diversity and functional potential varied in each study site. These results highlight the small-scale heterogeneity of geomorphologically comparable sites, greatly restricting generalizations about the fate of permafrost-affected environments in a warming Arctic.

M1476I, a French Canadian founder mutation of Na⁺ channel Nav1.4, causes potassium-aggravated myotonia, with cold-induced myotonia as the most distinctive clinical feature. Mexiletine, a class 1B local anaesthetic, relieves the myotonic symptoms of patients carrying the M1476I mutation. We used the patch-clamp method to investigate the functional characteristics of this mutation by heterologous expression in tsA201 cells. The M1476I mutation caused an increased persistent Na⁺ current, a 2- to 3-fold slower fast inactivation, a 6.4 mV depolarizing shift in the midpoint of steady-state inactivation, and an accelerated recovery from fast inactivation compared to the wild-type (WT) channel. Cooling slowed the kinetics of both channel types and increased the amplitude of the persistent current in M1476I channels.Mexiletine suppressed the persistent Na⁺ current generated by the M1476I mutation and blocked both WT and M1476I channels in a use- dependent manner. The inactivation-deficient M1476I channels were less susceptible to mexiletine during repetitive pulses. The decreased use-dependent block of M1476I channels might have resulted from the slower onset of mexiletine block, and/or the faster recovery from mexiletine block, given that the affinity of mexiletine for the inactivated state of the WT and mutant channels was similar. Increased extracellular concentrations of potassium had no effect on either M1476I or WT currents. These results indicated that cooling can augment the disruption of the voltage dependence of fast inactivation by M1476I channels.

Attached growth biological treatment systems are a promising solution to ammonia removal in cold-temperature climates. Environmental scanning electron microscopy (ESEM) and confocal laser scanning microscopy in combination with fluorescent in situ hybridization (FISH) was used to investigate the effects of 4 months of exposure to 4 degrees C on nitrifying biofilm and biomass. These molecular and microscopic methods were modified to minimize loss of mass and distortion of in situ perspectives. Environmental scanning electron microscopy revealed that nitrifying biofilm did not exhibit significant changes in volume with exposure to 4 degrees C. Confocal laser scanning microscopy in combination with FISH showed that the number of ammonia-oxidizing bacteria (AOB) cells present in the biofilm was statistically consistent during exposure to 4 degrees C. The RNA content of AOB cells remained sufficient for FISH enumeration. The number of nitrite-oxidizing bacteria cells remained steady during exposure to 4 degrees C; however, the RNA content of the cells appeared to decrease with exposure to 4 degrees C, thereby preventing their enumeration using FISH.

The temperature of packaged lettuce was recorded throughout a retail supply chain in Canada during the various stages of storage and shipping from the processor to retail. Temperatures were monitored in 27 cases of lettuce destined for three stores in three replicate trials conducted during the winter. A dynamic model that predicts the effect of temperature on the growth or die-off of Escherichia coli O157:H7 in packaged fresh-cut lettuce was applied to simulate the behavior of E. coli O157:H7 in the system. Simulations were carried out using distributions to account for variation in the temperature parameter and the die-off coefficient of the dynamic growth/death model. The results indicate that there was a predicted overall mean decline in cell numbers of 0.983 log cfu g⁻¹ and that the extent of cell death was proportional to the total time spent in the cold chain. Slight growth was predicted in a few instances when the dynamic temperature was above the permissive temperature of 5°C. These results suggest that generally there would be little or no growth of E. coli O157:H7 in product maintained at the proper temperature in the chain. Moreover, the predicted decline in cell numbers at refrigeration temperatures suggests that storage at 5°C or below prior to consumption would reduce populations of the pathogen in fresh-cut lettuce.

Fusarium graminearum Schwabe is the primary cause of Fusarium head blight (FHB) in North America. Chemically distinct F. graminearum sub-populations can be identified based on the type or composition of deoxynivalenol (DON) mycotoxin derivatives, including 3-acetyl (3-ADON) and 15-acetyl (15-ADON). The evaluation of randomly selected 3-ADON and 15-ADON isolates, collected from spring wheat throughout Canada, was performed using thin layer chromatography (TLC), high-performance liquid chromatography (HPLC), ice-nucleation activity (INA), and heat and cold tolerance tests conducted within a temperature range of -70°C to 65°C. The results indicated that the 3-ADON sub-population, which is responsible for the highest disease severity and has rapidly displaced the 15-ADON sub-population, produces more DON and zearalenone (ZEA) than the 15-ADON sub-population when exposed to heat and cold. Following exposures (1 and 2 h) to extremely high or low temperatures, 3-ADON isolates exhibited faster mycelial growth than 15-ADON isolates. In addition, the warmest temperature at which INA activity occurred was in 3-ADON (-3.6°C) vs. 15-ADON (-5.1°C). Taken together, these features suggest that the newly emerging 3-ADON sub-population is more resilient than the resident 15-ADON sub-population. Overall, the differences between the two sub-populations could provide new insights into FHB epidemiology and if validated under field conditions, may provide important information for predicting future FHB epidemics.

Objective: Several traffic safety research techniques require researchers to separate crash-involved drivers into culpable and nonculpable. Nonculpable drivers are assumed to be randomly involved in crashes by external factors and to approximate a noncollision control population. If this is true, factors that increase crash risk should be found more often in culpable than in nonculpable drivers. Though a culpability scoring tool has been developed for research purposes, that tool does not adequately address winter driving conditions (Robertson and Drummer 1994). Moreover, traditional culpability scoring requires assessors to read and score individual collision reports. The purpose of this study is to develop and validate an automated, rule-based Canadian culpability scoring tool that is capable of rapidly scoring police crash reports from large administrative datasets., Methods: We used an iterative approach to develop and validate our tool. First, the Robertson-Drummer culpability scoring tool was modified to include the extensive police report data collected in the British Columbia Traffic Accident System (TAS) and to account for winter driving conditions. This was done in consultation with traffic safety experts. The scoring tool was automated, employing a rule-based decision model that avoids interpretation of free-text reports. The scoring tool was applied to 73 collisions (134 drivers). Two experts also reviewed these collisions and determined the culpability of each driver. Discrepant cases were discussed to understand why the scoring tool differed from the expert assessment and the scoring tool was modified accordingly. The final tool was compared with expert assessment on another sample of 96 crashes. The tool was also applied to a sample of 2086 crash-involved drivers with known blood alcohol concentrations (BACs) and the adjusted odds of culpability were calculated for several BAC ranges., Results: The final scoring tool included 7 factors and had content validity for traffic safety experts. It had excellent agreement with expert scoring on the first set of collisions (kappa = 0.83, 95% confidence interval [CI]: 0.75-0.91) and on the second set (kappa = 0.84, 95% CI: 0.77-0.92). When applied to crash-involved drivers with known BAC levels, the scoring tool exhibited predictive validity: the odds of culpability increased with higher BACs, consistent with the known dose effect of BAC on crash risk., Conclusions: We have developed an automated culpability scoring tool contextualized to Canadian driving conditions. This tool will allow road safety researchers to assess collision responsibility in large administrative data sets derived from police reports.

During the winter in Western Canada, broilers are routinely transported in ambient temperatures ranging from 0°C to -40°C, yet there is little research in this area. This study examined the physiology and behavior of broilers undergoing simulated transport at typical Western Canadian winter temperatures. Groups of 15 broilers aged 32 to 33 d were exposed to an air stream regulated to -5, -10, or -15°C. Birds were placed into a typical transport drawer. Following baseline observations, the drawer was placed into a test chamber where cold air was drawn past the birds for 3 h. Three replications were conducted at each temperature. The birds adjusted their position within the drawer based upon the temperature distribution within the drawer. In comparison to the baseline period, exposing the birds to a cold air stream caused them to avoid the front plane (P = 0.003) which was the coldest area within the drawer. The birds did not adjust their usage of the middle (P = 0.308) and rear (P = 0.640) planes, because these were the warmer areas within the drawer. The total amount of space the birds occupied within the drawer did not decrease when exposed to the test chamber (P = 0.669). The core body temperature (CBT) did not vary and was within the known normal range during the normal (P = 0.528), pre-chamber (P = 0.060), and post-chamber (P = 0.285) periods. The CBT of the birds significantly decreased during the in-chamber period (P < 0.001) and then increased during the lairage period (P < 0.001). The shrink loss (P = 0.981) and amount of time to resume feed consumption (P = 0.357) were not affected by exposing the birds to temperatures of -5°C and colder. Exposing birds to temperatures of -5°C and colder had a negative effect on the CBT of the birds. However, the birds demonstrated behaviors which mitigated the negative effect that cold exposure could have on their CBT.

Subglacial sediments sampled from beneath Robertson Glacier (RG), Alberta, Canada, were shown to harbor diverse assemblages of potential nitrifiers, nitrate reducers, and diazotrophs, as assessed by amoA, narG, and nifH gene biomarker diversity. Although archaeal amoA genes were detected, they were less abundant and less diverse than bacterial amoA, suggesting that bacteria are the predominant nitrifiers in RG sediments. Maximum nitrification and nitrate reduction rates in microcosms incubated at 4°C were 280 and 18.5 nmol of N per g of dry weight sediment per day, respectively, indicating the potential for these processes to occur in situ. Geochemical analyses of subglacial sediment pore waters and bulk subglacial meltwaters revealed low concentrations of inorganic and organic nitrogen compounds. These data, when coupled with a C/N atomic ratio of dissolved organic matter in subglacial pore waters of ~210, indicate that the sediment communities are N limited. This may reflect the combined biological activities of organic N mineralization, nitrification, and nitrate reduction. Despite evidence of N limitation and the detection of nifH, we were unable to detect biological nitrogen fixation activity in subglacial sediments. Collectively, the results presented here suggest a role for nitrification and nitrate reduction in sustaining microbial life in subglacial environments. Considering that ice currently covers 11% of the terrestrial landmass and has covered significantly greater portions of Earth at times in the past, the demonstration of nitrification and nitrate reduction in subglacial environments furthers our understanding of the potential for these environments to contribute to global biogeochemical cycles on glacial-interglacial timescales.

The hemlock looper, Lambdina fiscellaria, is an economically important insect pest of Canadian forests which overwinters as eggs. Although the hemlock looper causes extensive damages, no information on the mechanisms related to its cold tolerance is known. The objective of this study was to determine the effect of temperature and exposure duration on hemlock looper winter survival but also to identify seasonal supercooling capacity and cryoprotectant levels of three populations along a latitudinal gradient. As host plant may contribute to offspring overwintering success, cold tolerance of hemlock looper eggs from parents whose larvae were fed on three different tree species was also measured. Mean supercooling point (SCP) of hemlock looper eggs was lower than -30 °C from October through the following spring with values being as low as -47 °C in February. Trehalose was the most abundant sugar found in hemlock looper eggs with a peak concentration of 0.3 μg mg⁻¹ DW⁻¹. Glycerol, a polyol, was more often absent in eggs of the different populations and tree species tested in the study. When exposed to different temperature regimes for various periods of time, significant mortality of hemlock looper eggs occurred at higher temperatures than the mean SCP. Thus, hemlock looper could be considered as a chill tolerant species. No clear pattern of population and host plant effects on SCP and cryoprotectants was detected in this study. However, when exposed to different winter temperatures and exposure duration, hemlock looper from higher latitudes survived better (survival rates ranging between 0 and 89% at -20 °C) than those from lower latitudes (survival rates ranging between 0 and 56% at -20 °C). Our results may contribute to a better understanding of hemlock looper winter biology and thus facilitate predictions of outbreaks and range expansion., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Background: Previous studies have suggested that the presence of the vascular component of hand-arm vibration syndrome (HAVS) in the hands increases the risk of cold-induced vasospasm in the feet., Aims: To determine if objectively measured cold-induced vasospasm in the hands is a risk factor for objectively measured cold-induced vasospasm in the feet in workers being assessed for HAVS., Methods: The subjects were 191 male construction workers who had a standardized assessment for HAVS including cold provocation digital photocell plethysmography of the hands and feet to measure cold-induced vasospasm. Bivariate analysis and multinomial logistic regression were used to examine the association between plethysmographic findings in the feet and predictor variables including years worked in construction, occupation, current smoking, cold intolerance in the feet, the Stockholm vascular stage and plethysmographic findings in the hands., Results: Sixty-one (32%) subjects had non-severe vasospasm and 59 (31%) had severe vasospasm in the right foot with the corresponding values being 57(30%) and 62 (32%) in the left foot. Multinomial logistic regression indicated that the only statistically significant predictor of severe vasospasm in the feet was the presence of severe vasospasm in the hands (OR: 4.11, 95% CI: 1.60-10.6, P < 0.01 on the right side and OR: 4.97, 95% CI: 1.82-13.53, P < 0.01 on the left side). Multinomial logistic regression analysis did not indicate any statistically significant predictors of non-severe vasospasm in the feet., Conclusions: Workers assessed for HAVS frequently have cold-induced vasospasm of their feet. The main predictor of severe vasospastic foot abnormalities is severe cold-induced vasospasm in the hands.

We report the first microbiological characterization of a terrestrial methane seep in a cryo-environment in the form of an Arctic hypersaline (∼24% salinity), subzero (-5 °C), perennial spring, arising through thick permafrost in an area with an average annual air temperature of -15 °C. Bacterial and archaeal 16S rRNA gene clone libraries indicated a relatively low diversity of phylotypes within the spring sediment (Shannon index values of 1.65 and 1.39, respectively). Bacterial phylotypes were related to microorganisms such as Loktanella, Gillisia, Halomonas and Marinobacter spp. previously recovered from cold, saline habitats. A proportion of the bacterial phylotypes were cultured, including Marinobacter and Halomonas, with all isolates capable of growth at the in situ temperature (-5 °C). Archaeal phylotypes were related to signatures from hypersaline deep-sea methane-seep sediments and were dominated by the anaerobic methane group 1a (ANME-1a) clade of anaerobic methane oxidizing archaea. CARD-FISH analyses indicated that cells within the spring sediment consisted of ∼84.0% bacterial and 3.8% archaeal cells with ANME-1 cells accounting for most of the archaeal cells. The major gas discharging from the spring was methane (∼50%) with the low CH(4)/C(2+) ratio and hydrogen and carbon isotope signatures consistent with a thermogenic origin of the methane. Overall, this hypersaline, subzero environment supports a viable microbial community capable of activity at in situ temperature and where methane may behave as an energy and carbon source for sustaining anaerobic oxidation of methane-based microbial metabolism. This site also provides a model of how a methane seep can form in a cryo-environment as well as a mechanism for the hypothesized Martian methane plumes.

Microbial consortia may harbour an array of resistance mechanisms that facilitate survival under harsh conditions, including antifreeze and ice-nucleation proteins. Antifreeze proteins lower freezing points as well as inhibit the growth of large, potentially damaging ice crystals from small ice embryos. In contrast, ice-nucleation proteins prevent supercooling and allow ice formation at high, sub-zero temperatures. Psychrophiles and psychrotolerant microbes are typically sought in extremely cold environments. However, given that geography is unlikely to present an insurmountable barrier to microbial dispersal, we reasoned that species with low-temperature adaptations should also be present, although rare, in more temperate environments. In consequence, the challenge then becomes one of selecting for rare microbes present in a larger community. Following the introductory commentary, we demonstrate that both freeze-thaw survival and ice-affinity selection can be used to identify microbes, which demonstrate low-temperature resistance, from enrichments derived from temperate environments. Selection resulted in a drastic decrease in cell abundance and diversity, allowing the isolation of a subset of resistant microbes. Depending on the origin of the consortia, these resistant microbes demonstrated cross-tolerance to osmotic stress, or a high proportion of antifreeze and/or ice-nucleation protein activities. Both types of ice-associating proteins presumably facilitate microbial survival at low temperatures. These proteins, as well as molecules that maintain osmotic balance, are also of commercial interest, with applications in the food, energy and medical industries. In addition, the resistant phenotypes described here provide a glimpse into the breadth of strategies microbes use to survive and thrive at low temperatures.

Few simple labeling methods exist for simultaneous fluorescence and electron microscopy of bacteria and biofilms. Here we describe the synthesis, characterization, and application of fluorescent nanoparticle quantum dot (QD) conjugates to target microbial species, including difficult to label Gram-negative strains. These QD conjugates impart contrast for both environmental scanning electron microscopy (ESEM) and fluorescence microscopy, permitting observation of living and fixed bacteria and biofilms. We apply these probes for studying biofilms extracted from perennial cold springs in the Canadian High Arctic, which is a particularly challenging system. In these biofilms, sulfur-metabolizing bacteria live in close association with unusual sulfur mineral formations. Following simple labeling protocols with the QD conjugates, we are able to image these organisms in fully-hydrated samples and visualize their relationship to the sulfur minerals using both ESEM and fluorescence microscopy. We then use scanning transmission electron microscopy to observe precipitated sulfur around individual cells and within the biofilm lattice. All combined, this information sheds light on the possible mechanisms of biofilm and mineral structure formation. These new QD conjugates and techniques are highly transferable to many other microbiological applications, especially those involving Gram-negative bacteria, and can be used for correlated fluorescence and electron microscopy.

Perennially cold habitats are diminishing as a result of climate change; however, little is known of the diversity or biogeography of microbes that thrive in such environments. Here we use targeted 16S rRNA gene surveys to evaluate the global affinities of cold-dwelling cyanobacteria from lake, stream and ice communities living at the northern limit of High Arctic Canada. Pigment signature analysis by HPLC confirmed the dominance of cyanobacteria in the phototrophic communities of these High Arctic microbial mats, with associated populations of chlorophytes and chromophytes. Microscopic analysis of the cyanobacteria revealed a diverse assemblage of morphospecies grouping into orders Oscillatoriales, Nostocales and Chroococcales. The 16S rRNA gene sequences from six clone libraries grouped into a total of 24 ribotypes, with a diversity in each mat ranging from five ribotypes in ice-based communities to 14 in land-based pond communities. However, no significant differences in composition were observed between these two microbial mat systems. Based on clone-library and phylogenetic analysis, several of the High Arctic ribotypes were found to be >99% similar to Antarctic and alpine sequences, including to taxa previously considered endemic to Antarctica. Among the latter, one High Arctic sequence was found 99.8% similar to Leptolyngbya antarctica sequenced from the Larsemann Hills, Antarctica. More than 68% of all identified ribotypes at each site matched only cyanobacterial sequences from perennially cold terrestrial ecosystems, and were <97.5% similar to sequences from warmer environments. These results imply the global distribution of low-temperature cyanobacterial ecotypes throughout the cold terrestrial biosphere.

Vertical pipes comprised of loose silica and lined by chert nodules have been observed in the abandoned Mafeking Quarry in central Manitoba, Canada. Discovery of microfossils within these features of the same age as the carbonate host rock indicates that they are a dissolution/replacement structure rather than infill of karst features by younger sediments. These features occur on the low thermal maturity edge of the intercratonic Williston Basin, are not associated with any known tectonic or hydrothermal activity, and show no sign of localized discharge of high-temperature fluids. Modern low-temperature brine springs with silica-filled discharge channels occur nearby, which suggests the silica chimneys are relic spring channels. Geochemical models have shown that dissolution/replacement reactions would be expected due to mixing of brine spring water with shallow groundwater in the region. Results indicate that silica pipe features in the rock record cannot be assumed to be indicative of hydrothermal activity. At the same time, results increase the astrobiological significance of low-temperature siliceous deposits.

Ecological concerns about antifouling paints containing non-green tin and copper compounds have highlighted the need for environmentally friendly alternatives. We report here a field test conducted in estuarine waters over two months designed to evaluate the efficiency of a number of active natural and man-made chemical ingredients added into a silicon-polyurethane marine paint. Early steps of biofouling in cold seawater of the St. Lawrence Estuary (Canada) were observed. Analyses, including dry biomass, flow cytometry and spectrofluorimetry, demonstrated a short-term antibacterial action of chitosan-based paints although no significant anti-algal action was observed. Cuprous oxide paints were efficient against bacteria and algae invasion in the first two weeks, especially those with added organic biocides such as isothiazolone and copper pyrithione. However, the overall dry biomass and chlorophyll a content were similar for all chitosan-and copper-based paints after 63 days. Microscopic observations revealed variation in the highly diverse benthic diatom population including species Navicula, Melosira, Cocconeis, Nitshzcia, Fragilaria and Amphora. Results suggest no real long-term efficiency for tested antifouling paints and highlight a particular need for green antifouling ingredients that are active under northern estuarine conditions.

A strictly anaerobic, Gram-positive, psychrotolerant, endospore-forming bacterium (strain A121(T)) was isolated from a permafrost sample collected in the Canadian High Arctic. Phylogenetic analysis of the 16S rRNA gene sequence of strain A121(T) showed its affiliation with the group of psychrophilic and psychrotolerant members of cluster I of the genus Clostridium, Clostridium bowmanii DSM 14206(T) being the closest relative (sequence similarity 98.5 %). Levels of DNA-DNA relatedness between strain A121(T) and the type strains of phylogenetically related species ranged from 33 to 52 %. Strain A121(T) grew in PY broth at temperatures between 4 and 28 degrees C (optimum 15-20 degrees C), at pH 6.0-8.0 (optimum pH 6.5-7.2) and in NaCl concentrations of 0-10.0 % (optimum 0-2.0 %). The strain utilized a narrow range of carbohydrates as sources of carbon and energy, including glucose, fructose, trehalose, maltose and starch; it also hydrolysed gelatin. Predominant fatty acids were C(16 : 1) cis9, C(16 : 1) cis9 DMA, C(16 : 0) and C(14 : 0). The DNA G+C content was 31.5 mol%. On the basis of its overall genotypic and phenotypic characteristics, strain A121(T) is classified within a novel species of the genus Clostridium, Clostridium tagluense sp. nov. The type strain is A121(T) (=VKM B-2369(T) =DSM 17763(T)).

Isolates of 'Pseudomonas borealis' were recovered after ice-affinity selection of summer-collected soils. 'P. borealis' DL7 was further characterized and shown to have ice nucleation activity (INA), a property that allows the crystallization of ice at temperatures close to the melting point, effectively preventing the supercooling of water. INA was optimally detected after culturing at temperatures consistent with psychrophilic growth. The sequence encoding the 'P. borealis' ice nucleation protein (INP) was obtained using both PCR and chromosome walking. When expressed in Escherichia coli, the resulting inaPb recombinants had INA. The 'P. borealis' sequence, dubbed inaPb, is clearly related to previously cloned INP genes, but it shows greater divergence. Sequence analysis suggests that there are two opposite flat surfaces, one relatively hydrophobic that likely serves as an ice template, and the other that could function as a complementary face to facilitate interprotein interaction for ice-step formation.

A Gram-negative and obligately anaerobic marine bacterium, strain HAW-EB21(T), was isolated in a previous study from marine sediment from the Atlantic Ocean, near Halifax Harbor, Canada, and found to have the potential to degrade both hexahydro-1,3,5-trinitro-1,3,5-triazine and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine. In the present study, phylogenetic analyses showed that strain HAW-EB21(T) was only distantly related to the genera Propionigenium and Ilyobacter with 6.6-7.5 % and 8.2-10.5 % dissimilarity as measured by 16S rRNA and 23S rRNA gene sequence analyses, respectively. Strain HAW-EB21(T) displayed unique properties in being psychrotrophic (18.5 degrees C optimum) and unable to utilize any of the carbon substrates (succinate, l-tartrate, 3-hydroxybutyrate, quinate or shikimate) used for isolating members of the genera Propionigenium and Ilyobacter. Strain HAW-EB21(T) utilized glucose, fructose, maltose, N-acetyl-d-glucosamine, citrate, pyruvate, fumarate and Casitone as carbon sources and produced H(2) and acetate as the major fermentation products. Cells grown at 10 degrees C produced C(15 : 1) (30 %), C(16 : 1)omega7 (15 %) and C(16 : 0) (16 %) as major membrane fatty acids. The novel strain had a genomic DNA G+C content of 28.1 mol%, lower than the values of the genera Ilyobacter and Propionigenium. Based on the present results, the novel isolate is suggested to be a member of a new genus for which the name Psychrilyobacter atlanticus gen. nov., sp. nov. is proposed. The type strain of the type species is HAW-EB21(T) (=DSM 19335(T)=JCM 14977(T)).

Fields from which oil is produced by injection of sulfate-bearing water often exhibit an increase in sulfide concentration with time (souring). Nitrate added to the injection water lowers the sulfide concentration by the action of sulfide-oxidizing, nitrate-reducing bacteria (SO-NRB). However, the injected nitrate can also be reduced with oil organics by heterotrophic NRB (hNRB). Aqueous volatile fatty acids (VFAs; a mixture of acetate, propionate, and butyrate) are considered important electron donors in this regard. Injection and produced waters from a western Canadian oil field with a low in situ reservoir temperature (30 degrees C) had only 0.1-0.2 mM VFAs. Amendment of these waters with nitrate gave therefore only partial reduction. More nitrate was reduced when 2% (v/v) oil was added, with light oil giving more reduction than heavy oil. GC-MS analysis of in vitro degraded oils and electron balance considerations indicated that toluene served as the primary electron donor for nitrate reduction. The differences in the extent of nitrate reduction were thus related to the toluene content of the light and heavy oil (30 and 5 mM, respectively). Reduction of nitrate with sulfide by SO-NRB always preceded that with oil organics by hNRB, even though microbially catalyzed kinetics with either electron donor were similar. Inhibition of hNRB by sulfide is responsible for this phenomenon. Injected nitrate will thus initially be reduced with sulfide through the action of SO-NRB. However, once sulfide has been eliminated from the near-injection wellbore region, oil organics will be targeted by the action of hNRB. Hence, despite the kinetic advantage of SO-NRB, the nitrate dose required to eliminate sulfide from a reservoir depends on the concentration of hNRB-degradable oil organics, with toluene being the most important in the field under study. Because the toluene concentration is lower in heavy oilthan in light oil, nitrate injection into a heavy-oil-producing field of low temperature is more likely to succeed in containing souring.

The saline springs of Gypsum Hill in the Canadian high Arctic are a rare example of cold springs originating from deep groundwater and rising to the surface through thick permafrost. The heterotrophic bacteria and autotrophic sulfur-oxidizing bacteria (up to 40% of the total microbial community) isolated from the spring waters and sediments were classified into four phyla (Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria) based on 16S rRNA gene analysis; heterotrophic isolates were primarily psychrotolerant, salt-tolerant, facultative anaerobes. Some of the isolates contained genes for thiosulfate oxidation (soxB) and anoxygenic photosynthesis (pufM), possibly enabling the strains to better compete in these sulfur-rich environments subject to long periods of illumination in the Arctic summer. Although leucine uptake by the spring water microbial community was low, CO(2) uptake was relatively high under dark incubation, reinforcing the idea that primary production by chemoautotrophs is an important process in the springs. The small amounts of hydrocarbons in gases exsolving from the springs (0.38 to 0.51% CH(4)) were compositionally and isotopically consistent with microbial methanogenesis and possible methanotrophy. Anaerobic heterotrophic sulfur oxidation and aerobic autotrophic sulfur oxidation activities were demonstrated in sediment slurries. Overall, our results describe an active microbial community capable of sustainability in an extreme environment that experiences prolonged periods of continuous light or darkness, low temperatures, and moderate salinity, where life seems to rely on chemolithoautotrophy.

The springs at Gypsum Hill and Colour Peak on Axel Heiberg Island in the Canadian Arctic originate from deep salt aquifers and are among the few known examples of cold springs in thick permafrost on Earth. The springs discharge cold anoxic brines (7.5 to 15.8% salts), with a mean oxidoreduction potential of -325 mV, and contain high concentrations of sulfate and sulfide. We surveyed the microbial diversity in the sediments of seven springs by denaturing gradient gel electrophoresis (DGGE) and analyzing clone libraries of 16S rRNA genes amplified with Bacteria and Archaea-specific primers. Dendrogram analysis of the DGGE banding patterns divided the springs into two clusters based on their geographic origin. Bacterial 16S rRNA clone sequences from the Gypsum Hill library (spring GH-4) were classified into seven phyla (Actinobacteria, Bacteroidetes, Firmicutes, Gemmatimonadetes, Proteobacteria, Spirochaetes, and Verrucomicrobia); Deltaproteobacteria and Gammaproteobacteria sequences represented half of the clone library. Sequences related to Proteobacteria (82%), Firmicutes (9%), and Bacteroidetes (6%) constituted 97% of the bacterial clone library from Colour Peak (spring CP-1). Most GH-4 archaeal clone sequences (79%) were related to the Crenarchaeota while half of the CP-1 sequences were related to orders Halobacteriales and Methanosarcinales of the Euryarchaeota. Sequences related to the sulfur-oxidizing bacterium Thiomicrospira psychrophila dominated both the GH-4 (19%) and CP-1 (45%) bacterial libraries, and 56 to 76% of the bacterial sequences were from potential sulfur-metabolizing bacteria. These results suggest that the utilization and cycling of sulfur compounds may play a major role in the energy production and maintenance of microbial communities in these unique, cold environments.

Several native poplar species meet at the margins of their natural distributions in southern Alberta, Canada. In this semi-arid area, poplars are obligate riparian species but they occupy several intergrading ecoregions. Populus deltoides Bartr. ex Marsh predominates in the warmest and driest eastern prairie ecoregions; P. balsamifera L. occupies the cooler and wetter western parkland and montane ecoregions; and P. angustifolia James and hybrids between the species occur in the intermediate grassland ecoregions. We investigated stomatal characteristics of these poplars in 51 genotypes collected across the range of ecoregions and grown in a semi-arid common garden. Stomatal length differed among genotypes within species but did not differ among species, ranging from 19 to 22 microm. Total stomatal densities (adaxial plus abaxial) differed among genotypes within species but were similar among species (290-420 stomata mm(-2)). Single-surface stomatal densities differed among species and consequently, the ratio of adaxial:abaxial stomatal density also differed, ranging from 0.94 for P. deltoides to 0.27 for P. balsamifera, with intermediate stomatal density ratios in P. angustifolia and hybrids. In a subsequent study of a subset of the same genotypes, stomatal density was correlated with stomatal conductance (r2 = 0.75) and the conductance ratios differed among species in the same manner as the stomatal density ratios. We conclude that: (1) diverse poplar genotypes respond similarly to a semi-arid environment by producing comparatively small and dense stomata; (2) differences in stomatal density underlie differences in stomatal conductance and differences among species in stomatal density ratio or conductance ratio may reflect adaptation to climatic differences among ecoregions; and (3) there is substantial variation in stomatal characteristics within and among species and hybrids in this area that could be useful for the selection and breeding of poplars adapted to different climatic conditions.

Identification of the pollution sources and understanding the processes related to runoff generation and pollution transportation is effective for the water quality management and selection of the Best Management Practices. The ANNualized AGricultural Non-Point Source (AnnAGNPS) model was applied to a watershed in Southern Ontario to evaluate the hydrology and sediment component from the non-point sources. The model was run for two years (1998 to 1999); one year's data was used to calibrate and the second year's data was used for validation purposes. The model has under predicted runoff amount and over predicted the sediment yield. However, the simulated runoff and sediment yield compared fairly well with the observed data indicating that the model had an acceptable performance in simulation of runoff and sediment. The study is still in progress to assess its performance for estimation of TMDL and improvements needed for the model to use under Ontario conditions.

Algae from cold water (Canada) and warm water (China) were analyzed for their total lipid content, and for their fatty acid (FA) composition and content. The major findings are that FA from Canadian algae are generally richer in polyunsaturated FA (PUFA), with a higher n-3/n-6 FA ratio, and a higher degree of total unsaturation. The 18 C, 4 double bonds FA (18 : 4 stearidonic acid, morotic acid as synonym) was detected in greater amounts in cold water samples. The high levels of total PUFA, and especially of n-3 FA in Canadian algae, suggests their possible utilizations for nutritional purposes.

The relationship between photosynthesis and accumulated cold degree days (CDD) over the late growing season was examined at the shoot, ecosystem and landscape scales in a boreal cutover in eastern Canada predominated by black spruce (Picea mariana Mill. BSP), lowbush blueberry (Vaccinium angustifolium Ait.) and sheep laurel (Kalmia angustifolia L.). We calculated CDD as the sum of minimum daily temperatures below a 5 degrees C threshold. Light-saturated photosynthesis at the shoot level (A(max)) of black spruce and V. angustifolium decreased steadily with increasing CDD once temperatures below the CDD threshold value became frequent in mid-September, whereas K. angustifolia showed a more irregular pattern. Tissue acclimation played an important role in the decrease in A(max) as the season progressed, but only V. angustifolium showed decreasing foliar nitrogen concentrations. Based on eddy covariance flux tower data, maximum daily gross primary productivity (GPP(max)-tower) at the ecosystem level was more strongly related to CDD (r(2) = 0.59) than was maximum daily net ecosystem exchange (r(2) = 0.32). The GPP(max) was likely influenced by both tissue acclimation and the direct effects of changing temperatures and irradiances on physiological rates. Mean daily GPP, calculated for consecutive 8-day periods for a 25 km(2) area around the tower by the MODIS MOD17A2 Collection 4 satellite algorithm (GPP- MODIS), decreased more rapidly with increasing CDD than did GPP(max)-tower. Although GPP-MODIS was closely correlated with mean daily GPP from the tower (GPP(daily)-tower, r(2) = 0.95) over the late growing season, the former was about twice as high. Although MODIS estimates of air temperature closely tracked the ground data, the maximum light-use efficiency parameter used by the MODIS algorithm was much higher than that indicated by the tower measurements. There was a 3% decline in GPP(max)-tower with an increase of 10 CDD, corresponding to the percent decline in branch-level A(max) of black spruce and V. angustifolium.

Independent studies were performed in Canada and in Denmark to assess the survival of parasitic wasps (Hymenoptera: Pteromalidae) wintering in puparia of house fly, Musca domestica Linnaeus (Diptera: Muscidae). Data in Canada were collected for Muscidifurax raptorGirault & Saunders, M. raptorellus Kogan & Legner, M. zaraptor Kogan & Legner, Nasonia vitripennis(Walker), Spalangia cameroni Perkins, Trichomalopsis sarcophagae (Gahan) and Urolepis rufipes (Ashmead) in three microsites at an outdoor cattle facility in southern Alberta. Survival was highest for N. vitripennis, T. sarcophagae and U. rufipes, ranging from near zero to c. 7%. No survival was observed for S. cameroni. Daily mean values for ambient air temperature (DMAT) averaged about -3.5 degrees C during exposure periods. Data for Denmark were collected for M. raptor, S. cameroni and U. rufipes in a dairy barn and in a swine barn. Survival of M. raptor and U. rufipes was higher than that of S. cameroni in the dairy barn (DMAT = 8.6 degrees C), with the three species having similar survival in the swine barn (DMAT = 15.4 degrees C). In both studies, parasitoids in egg stages were least likely to survive. These results identify the potential for T. sarcophagae and U. rufipes to be commercialized for use in northern climates as biocontrol agents for nuisance flies, compare directly the cold-hardiness of commercialized species (i.e. all of the above species excluding T. sarcophagae and U. rufipes), and document the importance of microsite on winter survival.

Objectives: To determine the long-term efficacy of cooled thermotherapy in the treatment of lower urinary tract symptoms of clinical benign prostatic hyperplasia., Methods: A total of 541 men underwent cooled thermotherapy treatment in six multicenter studies in the United States, England, and Canada. Both fixed and random effects models were used to pool the data across the six studies. The treatment response was measured as the difference between the urinary tract symptoms at baseline versus those at 3, 12, 24, 36, and 48 months after therapy. The treatment response included changes in the American Urological Association Symptom Score (AUA symptom score), peak urinary flow rate in milliliters per second (Qmax), and quality of life (QOL)., Results: The baseline measures were comparable across the studies. At 3 months, the AUA symptom score had improved by a mean of 11.6 (55%), Qmax by a mean of 4.0 (51%), and QOL by a mean of 2.3 (53%). These changes persisted with only slight attenuation through 48 months (corresponding mean changes of 43%, 35%, and 50%). These changes were highly statistically significant (P <0.0001 to 0.01). An improvement of at least 25% was achieved for the AUA symptom score and QOL by more than 85% of men and by more than 65% of men for Qmax., Conclusions: This pooled analysis of six multicenter studies of cooled thermotherapy, involving 541 men, found highly significant improvements in AUA symptom score, Qmax, and QOL. The results were highly consistent across the studies. The improvements reflected changes from baseline values of 45% to 50% for AUA symptom score and QOL and 35% to 40% for Qmax at a follow-up duration up to 48 months after therapy. The level of improvement for all three measures remained high at 48 months, indicating that the response is durable.

The impact of temperature on nitrification in biological granular activated carbon (GAC) filters was evaluated in order to improve the understanding of the nitrification process in drinking water treatment. The study was conducted in a northern climate where very cold water temperatures (below 2 degrees C) prevail for extended periods and rapid shifts of temperature are frequent in the spring and fall. Ammonia removals were monitored and the fixed nitrifying biomass was measured using a method of potential nitrifying activity. The impact of temperature was evaluated on two different filter media: an opened superstructure wood-based activated carbon and a closed superstructure activated carbon-based on bituminous coal. The study was conducted at two levels: pilot scale (first-stage filters) and full-scale (second-stage filters) and the results indicate a strong temperature impact on nitrification activity. Ammonia removal capacities ranged from 40 to 90% in pilot filters, at temperatures above 10 degrees C, while more than 90% ammonia was removed in the full-scale filters for the same temperature range. At moderate temperatures (4-10 degrees C), the first stage pilot filters removed 10-40% of incoming ammonia for both media (opened and closed superstructure). In the full-scale filters, a difference between the two media in nitrification performances was observed at moderate temperatures: the ammonia removal rate in the opened superstructure support (more than 90%) was higher than in the closed superstructure support (45%). At low temperatures (below 4 degrees C) both media performed poorly. Ammonia removal capacities were below 30% in both pilot- and full-scale filters.

Four psychrotrophic strains, which grew at 4 degrees C but not at 37 degrees C, were isolated from Japanese oil-reservoir water (strains SIB1, SIC1, SIS1) and Canadian oil sands (strain CAB1). Strains SIB1, SIS1, and CAB1 had a maximum growth rate at 20 degrees C and grew to the highest cell densities at the cultivation temperature of 0-4 degrees C. Strain SIS1 was capable of growing even at -5 degrees C. The growth profile of strain SIC1 was rather similar to that of a mesophilic bacterium. Strains SIB1, SIC1, and SIS1 were identified as members of the genus Shewanella, and strain CAB1 was a member of the genus Arthrobacter. All these strains exhibited weak degradation ability against catechol, a hydroxylated aromatic hydrocarbon, and tributyrin. These strains are expected to be of potential use in the in situ bioremediation technology of hazardous hydrocarbons and esters under low-temperature conditions.