Deciphering the Role of WWTPs in Cold Environments as Hotspots for the Dissemination of Antibiotic Resistance Genes.

Cold environments are the most widespread extreme habitats in the world. However, the role of wastewater treatment plants (WWTPs) in the cryosphere as hotspots in antibiotic resistance dissemination has not been well established. Hence, a snapshot of the resistomes of WWTPs in cold environments, below 5 °C, was provided to elucidate their role in disseminating antibiotic resistance genes (ARGs) to the receiving waterbodies. The resistomes of two natural environments from the cold biosphere were also determined. Quantitative PCR analysis of the aadA, aadB, ampC, blaSHV, blaTEM, dfrA1, ermB, fosA, mecA, qnrS, and tetA(A) genes indicated strong prevalences of these genetic determinants in the selected environments, except for the mecA gene, which was not found in any of the samples. Notably, high abundances of the aadA, ermB, and tetA(A) genes were found in the influents and activated sludge, highlighting that WWTPs of the cryosphere are critical hotspots for disseminating ARGs, potentially worsening the resistance of bacteria to some of the most commonly prescribed antibiotics. Besides, the samples from non-disturbed cold environments had large quantities of ARGs, although their ARG profiles were highly dissimilar. Hence, the high prevalences of ARGs lend support to the fact that antibiotic resistance is a common issue worldwide, including environmentally fragile cold ecosystems.

Exposure to hot and cold environments activates neurons projecting from the paraventricular thalamic nucleus to brain regions related to approach and avoidance behaviors.

Although cool- and warm-seeking behaviors for behavioral thermoregulation are considered to be appetitive/approach and aversive/avoidance behaviors, the neuronal circuits mediating such behaviors remain to be elucidated. A growing body of evidence suggests that the paraventricular thalamic nucleus (PVT) is a key brain region in a neuronal circuit that mediates appetitive/approach and aversive/avoidance behaviors. In this study, to elucidate the neuronal circuits mediating behavioral thermoregulatory responses, we examined whether neuronal pathways from the PVT to the nucleus accumbens (NAc), bed nucleus of the stria terminalis (BNST), and central nucleus of the amygdala (CeA), which are brain regions implicated in mediating appetitive/approach and aversive/avoidance behaviors, are activated during exposure to hot (38°C) and cold (8°C) environments using c-Fos immunostaining and retrograde tracing. Our results showed activation of neuronal pathways from the PVT to the NAc, BNST, and CeA during exposure to hot and cold environments, suggesting that activation of these pathways may be involved in avoidance behaviors from hot and cold environments for behavioral thermoregulation.

Going to sleep with a full belly: Thermal substitution by specific dynamic action in shorebirds.

Many bird species occupy habitats where environmental temperatures fall well below their thermoneutral zone (TNZ), so they must deal with high energy costs of thermoregulation to keep in heat balance. In such circumstances, specific dynamic action (SDA) - also referred to as heat increment of feeding - could be used to substitute for these high thermoregulatory costs. If birds ingest food before going to roost in cold environments, the SDA will be beneficial as an energy-conserving mechanism by thermal substitution. We investigated the magnitude and duration of SDA in a small-sized shorebird, the dunlin Calidris alpina, while feeding on living prey. We simulated in the aviary the food availability of a semidiurnal tidal cycle, and calculated the thermal substitution by SDA below their TNZ at the beginning of the "high tide" (resting period), after feeding ad libitum during the "low tide" (feeding period). Within TNZ (25 °C), dunlins consumed 12% (2.15 kJ) of the gross energy intake in excess by the SDA, with a duration of ~95 min. At 10 °C, i.e. below the lower critical limit of TNZ, SDA magnitude and duration were reduced by 29% and 31%, respectively. The amount of food ingested significantly affected the duration and magnitude of SDA, as well as the dunlin's body temperature. Thermal substitution by SDA saved 11% of the dunlin's theoretical daily energy requirement during winter. This thermal substitution could be commonly used by birds going to roost in cold climates. Interacting with other different behavioral and/or physiological strategies would help to maintain lower energetic costs and enhance survival in cold environments.

Metagenomic Analysis of Subtidal Sediments from Polar and Subpolar Coastal Environments Highlights the Relevance of Anaerobic Hydrocarbon Degradation Processes.

In this work, we analyzed the community structure and metabolic potential of sediment microbial communities in high-latitude coastal environments subjected to low to moderate levels of chronic pollution. Subtidal sediments from four low-energy inlets located in polar and subpolar regions from both Hemispheres were analyzed using large-scale 16S rRNA gene and metagenomic sequencing. Communities showed high diversity (Shannon's index 6.8 to 10.2), with distinct phylogenetic structures (<40% shared taxa at the Phylum level among regions) but similar metabolic potential in terms of sequences assigned to KOs. Environmental factors (mainly salinity, temperature, and in less extent organic pollution) were drivers of both phylogenetic and functional traits. Bacterial taxa correlating with hydrocarbon pollution included families of anaerobic or facultative anaerobic lifestyle, such as Desulfuromonadaceae, Geobacteraceae, and Rhodocyclaceae. In accordance, biomarker genes for anaerobic hydrocarbon degradation (bamA, ebdA, bcrA, and bssA) were prevalent, only outnumbered by alkB, and their sequences were taxonomically binned to the same bacterial groups. BssA-assigned metagenomic sequences showed an extremely wide diversity distributed all along the phylogeny known for this gene, including bssA sensu stricto, nmsA, assA, and other clusters from poorly or not yet described variants. This work increases our understanding of microbial community patterns in cold coastal sediments, and highlights the relevance of anaerobic hydrocarbon degradation processes in subtidal environments.

Subtercola vilae sp. nov., a novel actinobacterium from an extremely high-altitude cold volcano lake in Chile.

A novel actinobacterium, strain DB165T, was isolated from cold waters of Llullaillaco Volcano Lake (6170 m asl) in Chile. Phylogenetic analysis based on 16S rRNA gene sequences identified strain DB165T as belonging to the genus Subtercola in the family Microbacteriaceae, sharing 97.4% of sequence similarity with Subtercola frigoramans DSM 13057T, 96.7% with Subtercola lobariae DSM 103962T, and 96.1% with Subtercola boreus DSM 13056T. The cells were observed to be Gram-positive, form rods with irregular morphology, and to grow best at 10-15 °C, pH 7 and in the absence of NaCl. The cross-linkage between the amino acids in its peptidoglycan is type B2γ; 2,4-diaminobutyric acid is the diagnostic diamino acid; the major respiratory quinones are MK-9 and MK-10; and the polar lipids consist of phosphatidylglycerol, diphosphatidylglycerol, 5 glycolipids, 2 phospholipids and 5 additional polar lipids. The fatty acid profile of DB165T (5% >) contains iso-C14:0, iso-C16:0, anteiso-C15:0, anteiso-C17:0, and the dimethylacetal iso-C16:0 DMA. The genomic DNA G+C content of strain DB165T was determined to be 65 mol%. Based on the phylogenetic, phenotypic, and chemotaxonomic analyses presented in this study, strain DB165T (= DSM 105013T = JCM 32044T) represents a new species in the genus Subtercola, for which the name Subtercola vilae sp. nov. is proposed.

Description of Dioszegia patagonica sp. nov., a novel carotenogenic yeast isolated from cold environments.

During a survey of carotenogenic yeasts from cold and oligotrophic environments in Patagonia, several yeasts of the genus Dioszegia (Tremellales, Agaricomycotina) were detected, including three strains that could not be assigned to any known taxa. Analyses of internal transcribed spacer and D1/D2 regions of the large subunit rRNA gene showed these strains are conspecific with several other strains found in the Italian Alps and in Antarctica soil. Phylogenetic analyses showed that 19 of these strains represent a novel yeast species of the genus Dioszegia. The name Dioszegia patagonica sp. nov. is proposed to accommodate these strains and CRUB 1147T (UFMG 195T=CBMAI 1564T=DBVPG 10618T=CBS 14901T; MycoBank MB 819782) was designated as the type strain. This Dioszegia species accumulates biotechnologically valuable compounds such as carotenoid pigments and mycosporines.

Designing suitable shoes to prevent foot frostbite through optimization of the geometric dimensions of the shoe and sock model.

To prevent frostbite in cold environments, proper dimensions and materials for different parts of shoes along with the optimal design of shoe geometry were investigated. Furthermore, the optimal geometry of shoes was computed using an optimization algorithm to provide maximum thermal protection for the foot while having the lowest weight. The results showed that the length of the shoe sole and the thickness of the sock are the most effective parameters in foot protection against frostbite. Using thicker socks, which only increased the weight by roughly 11%, enhanced the minimum foot temperature by more than 2.3 times. HIGHLIGHTSOptimal design of shoe geometry is used to prevent frostbite in cold environments.A model of a biothermal nonlinear model is developed for the barefoot.Length of the shoe sole and the thickness of the sock are the most effective parameters in protecting the foot against frostbite.For the selected weather conditions, the toes are most likely to have frostbite.The best shoe for the selected weather conditions is the shoe that has the highest amount of thermal insulation in the toe area.

Adaptation strategies of giant viruses to low-temperature marine ecosystems.

Microbes in marine ecosystems have evolved their gene content to thrive successfully in the cold. Although this process has been reasonably well studied in bacteria and selected eukaryotes, less is known about the impact of cold environments on the genomes of viruses that infect eukaryotes. Here, we analyzed cold adaptations in giant viruses (Nucleocytoviricota and Mirusviricota) from austral marine environments and compared them with their Arctic and temperate counterparts. We recovered giant virus metagenome-assembled genomes (98 Nucleocytoviricota and 12 Mirusviricota MAGs) from 61 newly sequenced metagenomes and metaviromes from sub-Antarctic Patagonian fjords and Antarctic seawater samples. When analyzing our data set alongside Antarctic and Arctic giant viruses MAGs already deposited in the Global Ocean Eukaryotic Viral (GOEV) database, we found that Antarctic and Arctic giant viruses predominantly inhabit sub-10°C environments, featuring a high proportion of unique phylotypes in each ecosystem. In contrast, giant viruses in Patagonian fjords were subject to broader temperature ranges and showed a lower degree of endemicity. However, despite differences in their distribution, giant viruses inhabiting low-temperature marine ecosystems evolved genomic cold-adaptation strategies that led to changes in genetic functions and amino acid frequencies that ultimately affect both gene content and protein structure. Such changes seem to be absent in their mesophilic counterparts. The uniqueness of these cold-adapted marine giant viruses may now be threatened by climate change, leading to a potential reduction in their biodiversity.

Northern Pulmonary Hypertension: A Forgotten Kind of Pulmonary Circulation Pathology.

Northern pulmonary hypertension (NPH) is a medical condition that is still enigmatic in non-Russian-speaking countries. The extant previous literature is mostly available in the Russian language and, therefore, not accessible to the rest of the world. The recent increased interest in climate changes and environmental effects on pulmonary circulation prompted us to summarize the knowledge from the past about the effects of cold on pulmonary vasculature. In this review, we, for the first time, describe, in detail, the pathological attributes of human NPH, a medical disorder that occurs in people living in extremely cold regions, in the English language. Briefly, NPH is characterized by the hyper-muscularization of the pulmonary arteries and de novo muscularization of the arterioles with the ultimate development of right ventricular hypertrophy. However, the profound molecular mechanisms of the NPH pathology are to be revealed in future comprehensive studies.

Optimum design of the geometry of boots and socks with the aim of minimum weight and preventing frostbite.

In order to design shoes suitable for cold environments, knowledge of the thermal conditions inside the shoes and the variables affecting those conditions is necessary. A two-dimensional model of a boot and sock was developed to investigate the effect of the materials and dimensions of various parts of shoes and to design geometry for them to prevent foot frostbite. The optimization algorithm was used to optimize the dimensions of the boots to maximize the minimum foot temperature with the lowest boot weight. Two types of shoe soles and two kinds of shoe uppers were used to design suitable shoes. The results show the following: (1) In the design boots, the thermal insulation of the toe area plays an essential role in preventing frostbite. Two variables of the thickness of the toe cap and the length of the shoe sole had the greatest impact on the design of shoes with the least weight and the most protection against frostbite. So that to increase minimum foot temperature from 7°C to 15°C, 16°C, or 17°C, only the amounts of these variables should increase. (2) In designing the suitable boot, choosing the proper shoe sole had a significant effect on increasing the thermal insulation in the shoe and reducing its weight. So, for the boot with a minimum foot temperature of 20°C, by changing the shoe sole from EVA08 to EVA12, the weight is reduced by 42%. (3) To maximize the minimum foot temperature, it is necessary to use thick socks.

Fundamentals of Monitoring Condensation and Frost/Ice Formation in Cold Environments Using Thin-Film Surface-Acoustic-Wave Technology.

Moisture condensation, fogging, and frost or ice formation on structural surfaces cause severe hazards in many industrial components such as aircraft wings, electric power lines, and wind-turbine blades. Surface-acoustic-wave (SAW) technology, which is based on generating and monitoring acoustic waves propagating along structural surfaces, is one of the most promising techniques for monitoring, predicting, and also eliminating these hazards occurring on these surfaces in a cold environment. Monitoring condensation and frost/ice formation using SAW devices is challenging in practical scenarios including sleet, snow, cold rain, strong wind, and low pressure, and such a detection in various ambient conditions can be complex and requires consideration of various key influencing factors. Herein, the influences of various individual factors such as temperature, humidity, and water vapor pressure, as well as combined or multienvironmental dynamic factors, are investigated, all of which lead to either adsorption of water molecules, condensation, and/or frost/ice in a cold environment on the SAW devices. The influences of these parameters on the frequency shifts of the resonant SAW devices are systematically analyzed. Complemented with experimental studies and data from the literature, relationships among the frequency shifts and changes of temperature and other key factors influencing the dynamic phase transitions of water vapor on SAW devices are investigated to provide important guidance for icing detection and monitoring.

"Freezing" Thermophiles: From One Temperature Extreme to Another.

New detections of thermophiles in psychrobiotic (i.e., bearing cold-tolerant life forms) marine and terrestrial habitats including Arctic marine sediments, Antarctic accretion ice, permafrost, and elsewhere are continually being reported. These microorganisms present great opportunities for microbial ecologists to examine biogeographical processes for spore-formers and non-spore-formers alike, including dispersal histories connecting warm and cold biospheres. In this review, we examine different examples of thermophiles in cryobiotic locations, and highlight exploration of thermophiles at cold temperatures under laboratory conditions. The survival of thermophiles in psychrobiotic environments provokes novel considerations of physiological and molecular mechanisms underlying natural cryopreservation of microorganisms. Cultures of thermophiles maintained at low temperature may serve as a non-sporulating laboratory model for further exploration of metabolic potential of thermophiles at psychrobiotic temperatures, as well as for elucidating molecular mechanisms behind natural preservation and adaptation to psychrobiotic environments. These investigations are highly relevant for the search for life on other cold and icy planets in the Solar System, such as Mars, Europa and Enceladus.

Ubiquity and Diversity of Cold Adapted Denitrifying Bacteria Isolated From Diverse Antarctic Ecosystems.

Nitrogen cycle has been poorly investigated in Antarctic ecosystems. In particular, how extreme conditions of low temperature, dryness, and high radiation select the microorganisms involved in the cycle is not yet understood. Denitrification is an important step in the nitrogen cycle in which nitrate is reduced stepwise to the gases NO, N2O, and N2. Denitrification is carried out by a wide group of microorganisms spread in the phylogenetic tree. The aim of this work was to isolate and characterize denitrifying bacteria present in different cold environments from Antarctica. Bacterial isolates were obtained from lake, meltwater, sea, glacier ice, ornithogenic soil, and penguin feces samples from King George Island, Fildes peninsula in the Antarctic. Samples were taken during the deicing season in five sampling campaigns. From all the samples we were able to isolate denitrifying strains. A total of 199 bacterial isolates with the capacity to grow in anaerobic mineral media reducing nitrate at 4°C were obtained. The characterization of the isolates by 16S rRNA gene sequence analysis showed a high predominance of the genus Pseudomonas, followed by Janthinobacterium, Flavobacterium, Psychrobacter, and Yersinia. Other minor genera detected were Cryobacterium, Iodobacter, Kaistella, and Carnobacterium. The capacity to denitrify was not previously described for most of the bacteria related to our isolates and in many of them denitrifying genes were not present suggesting the presence of new genes in this extreme environment. Our work demonstrates the ubiquity of denitrification in the Maritime Antarctica and gives important information linking denitrification at cold temperature with taxa in an unequivocal way.

Low Ambient Temperature Exposition Impairs the Accuracy of a Non-invasive Heat-Flux Thermometer.

Background: Indirect core body temperature (CBT) monitoring from skin sensors is gaining attention for in-field applications thanks to non-invasivity, portability, and easy probe positioning. Among skin sensors, heat-flux devices, such as the so-called Double Sensor (DS), have demonstrated reliability under various experimental and clinical conditions. Still, their accuracy at low ambient temperatures is unknown. In this randomized cross-over trial, we tested the effects of cold temperature exposition on DS performance in tracking CBT. Methods: Twenty-one participants were exposed to a warm (23.2 ± 0.4°C) and cold (-18.7 ± 1.0°C) room condition for 10 min, following a randomized cross-over design. The accuracy of the DS to estimate CBT in both settings was assessed by quantitative comparison with esophageal (reference) and tympanic (comparator) thermometers, using Bland-Altman and correlation analyses (Pearson's correlation coefficient, r, and Lin's concordance correlation coefficient, CCC). Results: In the warm room setting, the DS showed a moderate agreement with the esophageal sensor [bias = 0.09 (-1.51; 1.69) °C, r = 0.40 (p = 0.069), CCC = 0.22 (-0.006; 0.43)] and tympanic sensor [bias = 2.74 (1.13; 4.35) °C, r = 0.54 (p < 0.05), CCC = 0.09 (0.008; 0.16)]. DS accuracy significantly deteriorated in the cold room setting, where DS temperature overestimated esophageal temperature [bias = 2.16 (-0.89; 5.22) °C, r = 0.02 (0.94), CCC = 0.002 (-0.05; 0.06)]. Previous exposition to the cold influenced temperature values measured by the DS in the warm room setting, where significant differences (p < 0.00001) in DS temperature were observed between randomization groups. Conclusion: DS accuracy is influenced by environmental conditions and previous exposure to cold settings. These results suggest the present inadequacy of the DS device for in-field applications in low-temperature environments and advocate further technological advancements and proper sensor insulation to improve performance in these conditions.

Ecology and potential functions of plant-associated microbial communities in cold environments.

Complex microbial communities are associated with plants and can improve their resilience under harsh environmental conditions. In particular, plants and their associated communities have developed complex adaptation strategies against cold stress. Although changes in plant-associated microbial community structure have been analysed in different cold regions, scarce information is available on possible common taxonomic and functional features of microbial communities across cold environments. In this review, we discuss recent advances in taxonomic and functional characterization of plant-associated microbial communities in three main cold regions, such as alpine, Arctic and Antarctica environments. Culture-independent and culture-dependent approaches are analysed, in order to highlight the main factors affecting the taxonomic structure of plant-associated communities in cold environments. Moreover, biotechnological applications of plant-associated microorganisms from cold environments are proposed for agriculture, industry and medicine, according to biological functions and cold adaptation strategies of bacteria and fungi. Although further functional studies may improve our knowledge, the existing literature suggest that plants growing in cold environments harbor complex, host-specific and cold-adapted microbial communities, which may play key functional roles in plant growth and survival under cold conditions.

Phytophthora species isolated from alpine and sub-alpine regions of Australia, including the description of two new species; Phytophthora cacuminis sp. nov and Phytophthora oreophila sp. nov.

Plant deaths had been observed in the sub-alpine and alpine areas of Australia. Although no detailed aetiology was established, patches of dying vegetation and progressive thinning of canopy suggested the involvement of root pathogens. Baiting of roots and associated rhizosphere soil from surveys conducted in mountainous regions New South Wales and Tasmania resulted in the isolation of eight Phytophthora species; Phytophthora cactorum, Phytophthora cryptogea, Phytophthora fallax, Phytophthora gonapodyides, Phytophthora gregata, Phytophthora pseudocryptogea, and two new species, Phytophthora cacuminis sp. nov and Phytophthora oreophila sp. nov, described here. P. cacuminis sp. nov is closely related to P. fallax, and was isolated from asymptomatic Eucalyptus coccifera and species from the family Proteaceae in Mount Field NP in Tasmania. P. oreophila sp. nov, was isolated from a disturbed alpine herbfield in Kosciuzsko National Park. The low cardinal temperature for growth of the new species suggest they are well adapted to survive under these conditions, and should be regarded as potential threats to the diverse flora of sub-alpine/alpine ecosystems. P. gregata and P. cryptogea have already been implicated in poor plant health. Tests on a range of alpine/subalpine plant species are now needed to determine their pathogenicity, host range and invasive potential.

Reproductive strategies in males of the world's southernmost lizards.

Reproductive and life history patterns in reptiles are tightly related to the environmental conditions, so male reproductive cycles have been historically characterized as continuous, for tropical lizards, or seasonal, for temperate lizards. However, males of Liolaemus and Phymaturus lizards (Liolaemidae), from cold temperate climates of high altitudes or latitudes in Argentina and Chile, have developed a variety of reproductive cycles to coordinate with the short female reproductive season and to deal with the low frequency of reproductive females in the population. Using gonadal histology and morphological analysis, we describe the male reproductive biology, fat storage and sexual dimorphism of the viviparous lizards Liolaemus sarmientoi and Liolaemus magellanicus that inhabit an austral grass steppe at 51°S, in the southern limit of the American continent. Males of L. sarmientoi and L. magellanicus are reproductively available during the entire activity season of approximately 5 months. In addition, males of both species exhibit greater body sizes than females in morphological variables relevant in sexual selection. Meanwhile, females of both species exhibit larger inter-limb length than conspecific males, which suggests fecundity selection to increase space for a larger litter size. The continuous sperm production throughout the activity season allows these liolaemids to mate at any time when females ovulate, representing a selective advantage to deal with the short activity season and the adversities of the cold environment they inhabit.

Atmospheric methane oxidizers are present and active in Canadian high Arctic soils.

The melting of permafrost and the associated potential for methane emissions to the atmosphere are major concerns in the context of global warming. However, soils can also represent a significant sink for methane through the activity of methane-oxidizing bacteria (MOB). In this study, we looked at the activity, diversity, and community structure of MOB at two sampling depths within the active layer in three soils from the Canadian high Arctic. These soils had the capacity to oxidize methane at low (15 ppm) and high (1000 ppm) methane concentrations, but rates differed greatly depending on the sampling date, depth, and site. The pmoA gene sequences related to two genotypes of uncultured MOB involved in atmospheric methane oxidation, the 'upland soil cluster gamma' and the 'upland soil cluster alpha', were detected in soils with near neutral and acidic pH, respectively. Other groups of MOB, including Type I methanotrophs and the 'Cluster 1' genotype, were also detected, indicating a broader diversity of MOB than previously reported for Arctic soils. Overall, the results reported here showed that methane oxidation at both low and high methane concentrations occurs in high Arctic soils and revealed that different groups of atmospheric MOB inhabit these soils.

Enhancing nitrification at low temperature with zeolite in a mining operations retention pond.

Ammonium nitrate explosives are used in mining operations at Diavik Diamond Mines Inc. in the Northwest Territories, Canada. Residual nitrogen is washed into the mine pit and piped to a nearby retention pond where its removal is accomplished by microbial activity prior to a final water treatment step and release into the sub-Arctic lake, Lac de Gras. Microbial removal of ammonium in the retention pond is rapid during the brief ice-free summer, but often slows under ice cover that persists up to 9 months of the year. The aluminosilicate mineral zeolite was tested as an additive to retention pond water to increase rates of ammonium removal at 4°C. Water samples were collected across the length of the retention pond monthly over a year. The structure of the microbial community (bacteria, archaea, and eukarya), as determined by denaturing gradient gel electrophoresis of PCR-amplified small subunit ribosomal RNA genes, was more stable during cold months than during July-September, when there was a marked phytoplankton bloom. Of the ammonia-oxidizing community, only bacterial amoA genes were consistently detected. Zeolite (10 g) was added to retention pond water (100 mL) amended with 5 mM ammonium and incubated at 12°C to encourage development of a nitrifying biofilm. The biofilm community was composed of different amoA phylotypes from those identified in gene clone libraries of native water samples. Zeolite biofilm was added to fresh water samples collected at different times of the year, resulting in a significant increase in laboratory measurements of potential nitrification activity at 4°C. A significant positive correlation between the amount of zeolite biofilm and potential nitrification activity was observed; rates were unaffected in incubations containing 1-20 mM ammonium. Addition of zeolite to retention ponds in cold environments could effectively increase nitrification rates year-round by concentrating active nitrifying biomass.