Characterization of gelatin-oxidized riclin cryogels and their applications as reusable ice cubes in shrimp preservation.

Traditional ice is usually employed to preserve food freshness and extend shelf life. However, ice cannot bear repeated freeze - thaw cycles during the transportation and retailing process, resulting in microbial cross-contamination and spoilage of foods. Herein, succinoglycan riclin was oxidated (RO) and crosslinked with gelatin (Ge), the Ge-RO cryogels were prepared via Schiff base reaction and three freeze - thaw cycles. The Ge-RO cryogels showed improved storage modulus (G') and thermal stability compared with pure gelatin hydrogel. The polymer framework of Ge-RO gels exhibited stable properties against ice crystals destructions during nine freeze - thaw treatments. During the storage and repeated freeze - thaw treatments of shrimps, Ge-RO cryogels exhibited a remarkable preservation effect on shrimps, and their freshness was evaluated using an electronic nose technique equipped with ten sensors. The results demonstrated that the shrimp muscle preserved in ice generated off-odors and resulted in high sensor responses. The sensor responses were reduced sharply of shrimps preserved in cryogels. Moreover, 1H NMR-based metabolomics analysis revealed that shrimps in Ge-RO cryogels group reversed the metabolic perturbations compared with the traditional ice group, the metabolic pathways were related to energy metabolism, nucleotide metabolism, and amino acid metabolism, which provide new clues to the freshness of shrimps. Furthermore, RO exhibited superior antimicrobial activity against E. coli and S. aureus microorganisms. Thus, the crosslinked cryogels are potentially applicable to food preservation, offering sustainable and reusable solutions against traditional ice.

Compromised heat loss leads to a delayed ice slurry induced reduction in heat storage.

Compromised heat loss due to limited convection and evaporation can increase thermal strain. We aimed to determine the effectiveness of ice slurry ingestion to reduce thermal strain following hyperthermia in a state of compromised heat loss. Twelve healthy males (age: 25 ± 4y) underwent hot water immersion to elevate rectal temperature (Trec) by 1.82 ± 0.08°C on four occasions. In the subsequent 60-min of seated recovery, participants ingested either 6.8 g·kg-1 of ice slurry (-0.6°C) or control drink (37°C) in ambient conditions (21 ± 1°C, 39 ± 10% relative humidity), wearing either t-shirt and shorts (2 trials: ICE and CON) or a whole-body sweat suit (2 trials: ICE-SS and CON-SS). Trec and mean skin temperature (Tsk) were recorded and a two-compartment thermometry model of heat storage was calculated. Heat storage was lower in ICE compared with CON at 20-40min (p ≤ 0.044, d ≥ 0.88) and for ICE-SS compared with CON-SS at 40-60 min (p ≤ 0.012, d ≥ 0.93). Trec was lower in ICE compared with CON from 30-60min (p ≤ 0.034, d ≥ 0.65), with a trend for a reduced Trec in ICE-SS compared with CON-SS at 40min (p = 0.079, d = 0.60). A greater Tsk was found in ICE-SS and CON-SS compared with ICE and CON (p < 0.001, d ≥ 3.37). A trend for a lower Tsk for ICE compared with CON was found at 20-40min (p ≤ 0.099, d ≥ 0.53), no differences were found for ICE-SS vs CON-SS (p ≥ 0.554, d ≤ 0.43). Ice slurry ingestion can effectively reduce heat storage when heat loss through convection and evaporation is compromised, relevant to those wearing personal protective equipment or those with compromised sweat loss. Compromised heat loss delays the reduction in heat storage, possibly related to ice slurry ingestion not lowering Tsk.

Ice formation and its elimination in cryopreservation of oocytes.

Damage from ice and potential toxicity of ice-inhibiting cryoprotective agents (CPAs) are key issues in assisted reproduction of humans, domestic and research animals, and endangered species using cryopreserved oocytes and embryos. The nature of ice formed in bovine oocytes (similar in size to oocytes of humans and most other mammals) after rapid cooling and during rapid warming was examined using synchrotron-based time-resolved x-ray diffraction. Using cooling rates, warming rates and CPA concentrations of current practice, oocytes show no ice after cooling but always develop large ice fractions-consistent with crystallization of most free water-during warming, so most ice-related damage must occur during warming. The detailed behavior of ice at warming depended on the nature of ice formed during cooling. Increasing cooling rates allows oocytes soaked as in current practice to remain essentially ice free during both cooling and warming. Much larger convective warming rates are demonstrated and will allow routine ice-free cryopreservation with smaller CPA concentrations. These results clarify the roles of cooling, warming, and CPA concentration in generating ice in oocytes and establish the structure and grain size of ice formed. Ice formation can be eliminated as a factor affecting post-warming oocyte viability and development in many species, improving outcomes and allowing other deleterious effects of the cryopreservation cycle to be independently studied.

The water and oridonin sources in the ice ribbons of Isodon rubescens.

To study the source and content change of oridonin in the ice ribbons, the contents of oridonin in the ice ribbons and bleeding sap of Isodon rubescens at different times were determined with RP-HPLC. The paraffin sectioning and electron microscopy imaging were performed to study the transport channel of oridonin in the stem. The results showed that there were abundant xylem rays and perfect pit pairs in the secondary xylem of I. rubescens stems. The oridonin content in the ice ribbons of I. rubescens stems was lower than that in the stem of I. rubescens and even decreased over time. The contents of oridonin in the bleeding sap of I. rubescens stems was equal to that in second-day ice ribbons and was lower than that in first-day ice ribbons. The water in the ice ribbons of I. rubescens stems originated from water absorbed by the roots from soil. This water was transported from the roots of I. rubescens to the stem and then transferred through efficient lateral conducting tissues to the surface of the stem. The oridonin in the phloem and cortex of I. rubescens stems dissolves in water originating from the soil and freezes in the form of ice ribbons below 0 °C.

Microbial ice-binding structures: A review of their applications.

Microorganisms' ice-binding structures (IBS) are macromolecules with potential commercial value in agriculture, food technology, material technology, cryobiology, and medicine. Microbial ice-structuring or microbial ice-binding particles, with their multi-applications, are simple to use, effective in low amounts, non-toxic, and environmentally friendly. Due to their source and composition diversities, microbial ice-binding structures are gaining attention because they are useable in various conditions. Some microorganisms also produce structures with dual ice-nucleating and anti-freezing properties. Structures that promote ice formation (ice nucleating particles- INPs) act as ice nuclei, lowering the energy barrier between supercooled liquid and ice, causing ice crystals to form. In contrast, anti-freeze particles (AFPs) prevent ice formation and recrystallization through several mechanisms, including disturbing the formation of string hydrogen bonds amongst water molecules, melting already formed ice crystals, and preventing crystal formation by binding to specific sites. Knowledge of the type and function of microbial ice-binding structures lends fundamental insight for possible scaling the production of cheap, functional, and advanced microbial structure-inspired mimics and by-products. This review focuses on microbial ice-binding structures and their potential uses in the food, medicinal, environmental, and agricultural sectors.

Nourishing Physical Productivity and Performance On a Warming Planet - Challenges and Nutritional Strategies to Mitigate Exertional Heat Stress.

PURPOSE OF REVIEW: Climate change is predicted to increase the frequency and severity of exposure to hot environments. This can impair health, physical performance, and productivity for active individuals in occupational and athletic settings. This review summarizes current knowledge and recent advancements in nutritional strategies to minimize the impact of exertional-heat stress (EHS). RECENT FINDINGS: Hydration strategies limiting body mass loss to < 3% during EHS are performance-beneficial in weight-supported activities, although evidence regarding smaller fluid deficits (< 2% body mass loss) and weight-dependent activities is less clear due to a lack of well-designed studies with adequate blinding. Sodium replacement requirements during EHS depends on both sweat losses and the extent of fluid replacement, with quantified sodium replacement only necessary once fluid replacement > 60-80% of losses. Ice ingestion lowers core temperature and may improve thermal comfort and performance outcomes when consumed before, but less so during activity. Prevention and management of gastrointestinal disturbances during EHS should focus on high carbohydrate but low FODMAP availability before and during exercise, frequent provision of carbohydrate and/or protein during exercise, adequate hydration, and body temperature regulation. Evidence for these approaches is lacking in occupational settings. Acute kidney injury is a potential concern resulting from inadequate fluid replacement during and post-EHS, and emerging evidence suggests that repeated exposures may increase the risk of developing chronic kidney disease. Nutritional strategies can help regulate hydration, body temperature, and gastrointestinal status during EHS. Doing so minimizes the impact of EHS on health and safety and optimizes productivity and performance outcomes on a warming planet.

Radiolytic Effects on Biological and Abiotic Amino Acids in Shallow Subsurface Ices on Europa and Enceladus.

Europa and Enceladus are key targets to search for evidence of life in our solar system. However, the surface and shallow subsurface of both airless icy moons are constantly bombarded by ionizing radiation that could degrade chemical biosignatures. Therefore, sampling of icy surfaces in future life detection missions to Europa and Enceladus requires a clear understanding of the necessary ice depth where unaltered organic biomolecules might be present. We conducted radiolysis experiments by exposing individual amino acids in ices and amino acids from dead microorganisms in ices to gamma radiation to simulate conditions on these icy worlds. In the pure amino acid samples, glycine did not show a detectable decrease in abundance, whereas the abundance of isovaline decreased by 40% after 4 MGy of exposure. Amino acids in dead Escherichia coli (E. coli) organic matter exhibited a gradual decline in abundances with the increase of exposure dosage, although at much slower rates than individual amino acids. The majority of amino acids in dead A. woodii samples demonstrated a step function decline as opposed to a gradual decline. After the initial drop in abundance with 1 MGy of exposure, those amino acids did not display further decreases in abundance after exposure up to 4 MGy. New radiolysis constants for isolated amino acids and amino acids in dead E. coli material for Europa/Enceladus-like conditions have been derived. Slow rates of amino acid destruction in biological samples under Europa and Enceladus-like surface conditions bolster the case for future life detection measurements by Europa and Enceladus lander missions. Based on our measurements, the "safe" sampling depth on Europa is ∼20 cm at high latitudes of the trailing hemisphere in the area of little impact gardening. Subsurface sampling is not required for the detection of amino acids on Enceladus-these molecules will survive radiolysis at any location on the Enceladus surface. If the stability of amino acids observed in A. woodii organic materials is confirmed in other microorganisms, then the survival of amino acids from a potential biosphere in Europa ice would be significantly increased.

High-speed cryo-microscopy reveals that ice-nucleating proteins of Pseudomonas syringae trigger freezing at hydrophobic interfaces.

Ice-nucleating proteins (INpro) trigger the freezing of supercooled water droplets relevant to atmospheric, biological, and technological applications. The high ice nucleation activity of INpro isolated from the bacteria Pseudomonas syringae could be linked to the aggregation of proteins at the bacterial membrane or at the air-water interface (AWI) of droplets. Here, we imaged freezing onsets, providing direct evidence of these proposed mechanisms. High-speed cryo-microscopy identified the onset location of freezing in droplets between two protein-repellent glass slides. INpro from sterilized P. syringae (Snomax) statistically favored nucleation at the AWI of the droplets. Removing cellular fragments by filtration or adding surfactants increased the frequency of nucleation events at the AWI. On the other hand, cultivated intact bacteria cells or lipid-free droplets nucleated ice without an affinity to the AWI. Overall, we provide visual evidence that INpro from P. syringae trigger freezing at hydrophobic interfaces, such as the AWI or the bacterial membrane, with important mechanistic implications for applications of INpro.

Furcellaran: Impact of Concentration, Rheological Properties, and Structure on Ice Recrystallization Inhibition Activity.

Recrystallization is considered the main damaging mechanism during the frozen storage of biologic materials. In this study, furcellaran, a polysaccharide related to κ-carrageenan, was studied for its concentration-dependent effect on ice crystal growth and recrystallization. The structure and sulfate content of the utilized furcellaran was analyzed by 1H nuclear magnetic resonance spectroscopy, ion chromatography, and high-performance size-exclusion chromatography. Additionally, the rheological properties of furcellaran solutions were investigated. Our findings demonstrate that furcellaran inhibits ice growth as effectively as κ-carrageenan. Furthermore, the rheological properties change with increasing furcellaran concentration, resulting in a gel-like consistency at 5 g/L, which coincides with decreased recrystallization inhibition activity and larger crystals. This suggests that gel formation or a gel-like consistency has to be avoided for optimal recrystallization inhibition activity.

Bacterial diversity and geomicrobiology of Winter Wonderland ice cave, Utah, USA.

The Winter Wonderland ice cave, located at an elevation of 3140 m above sea level in the Uinta Mountains of northern Utah, USA, maintains a constant sub-zero temperature. Seasonal snowmelt and rain enter the cave, freeze on the surface of the existing ice, and contribute to a 3-m-thick layered ice mass. This ice mass contains organic matter and cryogenic cave carbonates (CCCs) that date back centuries. In this study, samples of ice, liquid water, and exposed CCCs were collected to examine the bacterial communities within the cave and to determine if these communities vary spatially and between sample types. Flow cytometry showed that cell counts are an order of magnitude higher in liquid water samples than in ice. Epifluorescence microscopy and scanning electron microscopy imaging revealed potential coccoid and bacillus microbial morphologies in water samples and putative cells or calcite spherules in the CCCs. The diversity of bacteria associated with soil, identified through sequence-based analysis, supports the hypothesis that water enters the cave by filtering through soil and bedrock. A differential abundance of bacterial taxa was observed between sample types, with the greatest diversity found in CCCs. This supports a geomicrobiological framework where microbes aggregate in the water, sink into a concentrated layer, and precipitate out of the ice with the CCCs, thereby reducing the cell counts in the ice. These CCCs may provide essential nutrients for the bacteria or could themselves be products of biomineralization.

In situ investigation of ice fractions and water states during partial freezing of pork loins and shrimps.

Ice fractions and water states in partially frozen muscle foods greatly affect their quality. In the study, a variable temperature nuclear magnetic resonance (VT-NMR) with a liquid nitrogen temperature control system was employed to in situ investigate the relationship between ice fractions and temperatures and changes in water states during partial freezing and thawing of pork and shrimp. Results indicated that changes in ice fractions ranging from -2 âˆ¼ -20 °C could be divided into 3 stages including slow increase, random leap and remarkable leap. More serious damages to the structures related to immobile water occurred in shrimp than in pork, and partial freezing also caused deterioration in muscle fibres related to free water. Additionally, -2 âˆ¼ -3 °C and - 3.5 °C were the appropriate partial freezing temperatures for pork and shrimp, respectively. Therefore, the VT-NMR method possessed great potential for fundamental studies and applications of partial freezing of muscle foods.

Freezing rate's impact on starch retrogradation, ice recrystallization, and quality of water-added and water-free quick-frozen rice noodles.

The study evaluated the effect of freezing rate on the quality of water-added quick-frozen rice noodles and water-free quick-frozen rice noodles. Results indicated that the retrogradation enthalpy, relative crystallinity, freezable water content, and cooking loss of water-added quick-frozen rice noodles were higher than those of water-free quick-frozen rice noodles with increasing storage time. Furthermore, ice recrystallization accelerated the deterioration of the quality of the rice noodles, resulting in the enlargement of the pores within the rice noodles and the formation of many pores on the surface. This phenomenon was particularly evident in the rice noodles of Y-40 °C (freezing with water at -40 °C) and Y-60 °C (freezing with water at -60 °C). After 28 days of frozen storage, the hardness increased by 83.83 % for rice noodles of Y-20 °C (freezing with water at -20 °C), while the hardness decreased by 51.68 % and 45.80 %, respectively, for rice noodles of Y-40 °C and Y-60 °C. Consequently, the impact of the freezing rate on the quality of water-added quick-frozen rice noodles is more pronounced than that of water-free quick-frozen rice noodles. Moreover, a higher freezing rate can delay the deterioration of the quality of frozen rice noodles by postponing starch retrogradation and inhibiting ice recrystallization.

Application of RiTiCE in understanding hydro-meteorological controls on ice break-up patterns in River Tornionjoki.

The Arctic region experiences significant annual hydrologic events, with the spring flood and ice break-up being the most prominent. River ice break-up, in particular, poses high socioeconomic and ecological expenses, including morphological changes and damage to riverine structures. This study aims to investigate the spatiotemporal patterns of river ice in the River Tornionjoki, including the timing of ice break-up at different latitudes. We utilized observation data and remote sensing techniques to track changes in ice patterns overtime on the River Tornionjoki. The study indicates that the ice break-up in the River Tornionjoki basin typically occurs during Apr-Jun based on the reach location in different latitudes; therefore, different stations behave according to their latitudinal location. We observed significant spatial variations in ice break-up timing across the basin, with an earlier break-up in the lower latitudes compared to the upper latitudes. The average ice break-up day in lower latitude stations ranges between 200-205, while in higher latitude stations the average ice break-up day ranges between 215-228.

Analysis of changes in the occurrence of ice phenomena in upland and mountain rivers of Poland.

The ice phenomena are an inherent component of rivers in temperate, continental, and polar climate zones. Evident progress in global warming leads to a decrease in snow cover on land and ice phenomena in water bodies, disrupting the stability of the hydrological cycle and aquatic ecosystems. Although common observations indicate the disappearance of ice phenomena in rivers over recent decades, detailed quantitative research is lacking in many regions, especially in the temperate zone. In this paper, ice phenomena were analyzed on the rivers of southern Poland, located in the upland and mountain areas of the country, as no such studies have been conducted so far. The temporal changes in the annual number of days with ice (NDI) phenomena were studied in locations where ice phenomena were observed every year for at least 30 years between 1951 and 2021. Using straightforward but commonly accepted procedures, such as the Mann-Kendall test, statistically significant decreasing trends in the annual NDI were revealed for the majority of gauging stations. The Theil-Sen (TS) slope mean values were -1.66 (ranging from -3.72 to -0.56), -1.41 (from -3.22 to -0.29), and -1.33 (from -2.85 to -0.29) for the datasets representing the periods 1992-2020, 1987-2020, and 1982-2020, respectively. The results for the annual NDI were additionally presented within the context of meteorological characteristics such as annual and winter (Nov-Apr) air temperature, precipitation, and water temperature. Correlation and regression analyses revealed that the main factor triggering the decrease in NDI is the increase in the average winter air temperature. An increase in temperature by 1°C results in a decrease in NDI by up to twenty days. If these negative trends continue, ice phenomena may disappear completely from southern Polish rivers within few decades.

Ice in the intertidal: patterns and processes of freeze tolerance in intertidal invertebrates.

Many intertidal invertebrates are freeze tolerant, meaning that they can survive ice formation within their body cavity. Freeze tolerance is a fascinating trait, and understanding its mechanisms is important for predicting the survival of intertidal animals during extreme cold weather events. In this Review, we bring together current research on the ecology, biochemistry and physiology of this group of freeze-tolerant organisms. We first introduce the ecology of the intertidal zone, then highlight the strong geographic and taxonomic biases within the current body of literature on this topic. Next, we detail current knowledge on the mechanisms of freeze tolerance used by intertidal invertebrates. Although the mechanisms of freeze tolerance in terrestrial arthropods have been well-explored, marine invertebrate freeze tolerance is less well understood and does not appear to work similarly because of the osmotic differences that come with living in seawater. Freeze tolerance mechanisms thought to be utilized by intertidal invertebrates include: (1) low molecular weight cryoprotectants, such as compatible osmolytes and anaerobic by-products; (2) high molecular weight cryoprotectants, such as ice-binding proteins; as well as (3) other molecular mechanisms involving heat shock proteins and aquaporins. Lastly, we describe untested hypotheses, methods and approaches that researchers can use to fill current knowledge gaps. Understanding the mechanisms and consequences of freeze tolerance in the intertidal zone has many important ecological implications, but also provides an opportunity to broaden our understanding of the mechanisms of freeze tolerance more generally.

Frost fighters: unveiling the potential of microbial antifreeze proteins in biotech innovation.

Polar environments pose extreme challenges for life due to low temperatures, limited water, high radiation, and frozen landscapes. Despite these harsh conditions, numerous macro and microorganisms have developed adaptive strategies to reduce the detrimental effects of extreme cold. A primary survival tactic involves avoiding or tolerating intra and extracellular freezing. Many organisms achieve this by maintaining a supercooled state by producing small organic compounds like sugars, glycerol, and amino acids, or through increasing solute concentration. Another approach is the synthesis of ice-binding proteins, specifically antifreeze proteins (AFPs), which hinder ice crystal growth below the melting point. This adaptation is crucial for preventing intracellular ice formation, which could be lethal, and ensuring the presence of liquid water around cells. AFPs have independently evolved in different species, exhibiting distinct thermal hysteresis and ice structuring properties. Beyond their ecological role, AFPs have garnered significant attention in biotechnology for potential applications in the food, agriculture, and pharmaceutical industries. This review aims to offer a thorough insight into the activity and impacts of AFPs on water, examining their significance in cold-adapted organisms, and exploring the diversity of microbial AFPs. Using a meta-analysis from cultivation-based and cultivation-independent data, we evaluate the correlation between AFP-producing microorganisms and cold environments. We also explore small and large-scale biotechnological applications of AFPs, providing a perspective for future research.

Frost-free zone on leaves revisited.

Particular frost patterns on natural leaves had prompted Yao et al. [Y. Yao et al., Proc. Natl. Acad. Sci. U.S.A. 117, 6323-6329 (2020)] to investigate the underlying physics. Their work revealed why on corrugated surfaces ice forms on crests and dries out adjacent grooves. In the absence of frost, in contrast, grooves tend to constitute niches on a leaf where microorganisms are less limited by moisture than in other locations. Here, we show that microorganisms able to nucleate ice before it forms on crests can modify the frosting pattern to their advantage. This ability might drive in cold arid environments the association between certain microorganisms and plants.

Ice nucleation active bacteria metabolites as antibiofilm agent to control Aeromonas hydrophila and Streptococcus agalactiae infections in Aquaculture.

OBJECTIVES: The aim of this study was to quantify and identify metabolites of Ice Nucleation Active (INA) bacteria as an anti-biofilm agent against biofilms of fish pathogens such as Aeromonas hydrophila and Streptococcus agalactiae. RESULTS: Ice nucleation active bacteria, which have the ability to catalyze ice nucleation, isolated from rainwater in previous studies, were used. All INA isolates were tested in several assays, including the antimicrobial test, which uses streptomycin as the positive control and none of the isolates were found positive in the antimicrobial test. As for the quorum quenching assay, it was found that four out of ten isolates were able to disturb the communication system in Chromobacterium violaceum wild type, which was used as the indicator bacteria. On the next assay, all ten isolates were tested for Biofilm Inhibition and Destruction and showed anti-biofilm activity with the highest percentage inhibition of 33.49% by isolate A40 against A. hydrophila and 77.26% by isolate A19 against S. agalactiae. C1 performed the highest destruction against A. hydrophila and S. agalactiae, with percentages of 32.11% and 51.88%, respectively. As for the GC-MS analysis, supernatants of INA bacteria contain bioactive compounds such as sarcosine and fatty acids, which are known to have antibiofilm activity against several biofilm-forming bacteria. Through 16s rRNA sequencing, identified bacteria are from the Pantoea, Enterobacter, and Acinetobacter genera. As for the conclusion, ice nucleation active bacteria metabolites tested showed positive results against pathogenic bacteria Aeromonas hydrophila and Streptococcus agalactiae in destructing and inhibiting biofilm growth.

On the engulfment of antifreeze proteins by ice.

Antifreeze proteins (AFPs) are remarkable biomolecules that suppress ice formation at trace concentrations. To inhibit ice growth, AFPs must not only bind to ice crystals, but also resist engulfment by ice. The highest supercooling, [Formula: see text], for which AFPs are able to resist engulfment is widely believed to scale as the inverse of the separation, [Formula: see text], between bound AFPs, whereas its dependence on the molecular characteristics of the AFP remains poorly understood. By using specialized molecular simulations and interfacial thermodynamics, here, we show that in contrast with conventional wisdom, [Formula: see text] scales as [Formula: see text] and not as [Formula: see text]. We further show that [Formula: see text] is proportional to AFP size and that diverse naturally occurring AFPs are optimal at resisting engulfment by ice. By facilitating the development of AFP structure-function relationships, we hope that our findings will pave the way for the rational design of AFPs.