Concentrations Of Mineral Salts Research Articles

Analysis of Total Plate Count Microba in Fresh (Rastrelliger sp.) Mackerel fish` of Sorong City West Papua

Fish is one of the fishery food ingredients that are needed by humans, because in the fish meat there are compounds needed by the body. Such as the content of protein compounds, fats, carbohydrates, vitamins and mineral salts. Sorong in the city of mackerel is one of the fish that are of interest to the community, and its availability is abundant at the fish auction place in the city of Sorong. But in general the consumption of mackerel has not been tested either chemically or microbiologically based on the background of the purpose of this study is to determine the total microbial plate count in fresh mackerel (Rastelliger sp.) In Sorong City, West Papua. The method in this research was descriptive method, which is drawing the number of microbes through the test of the total number of microbial plates (ALT). Based on the results of the study which stated the average of the five samples approved were in sample A and sample B the amount of ALT was 3.7 x 104 cFU/mL, sample C and sample D were 1.02 x 105 cFU/mL, and sample E as much as 2.7 x 105 cFU/mL. The total number of microbial total plates (ALT) or the number of microbial total plates (TPC) in fresh mackerel (Rastrelliger sp.) in the Sorong TPI city does not exceed the maximum Indonesian national standard (SNI), and it is concluded that the fish can be consumed.

Evaluation of biotechnological potential of thermotolerant microorganisms for oil degradation

The problem of pollution of natural ecosystems with oil and oil products is extremely acute in manycountries of the world, including Kazakhstan. A significant part of oil-contaminated sites is located inregions with a hot climate, and in such countries studies are underway to isolate thermotolerant oildegrading microorganisms with a view to their further use for remediation of oil-contaminated territories.The circle of such microorganisms is quite wide and diverse.The effectiveness of the method of bioremediation of oil-polluted ecosystems largely depends onbiotic and abiotic factors, for example, salinity, temperature, ability to produce surfactants, pH, etc.Earlier from the soils of Western Kazakhstan, we isolated and identified thermotolerant oil-oxidizing microorganisms capable of oxidizing oil hydrocarbons in a wide temperature range. The aim of this workwas to study the ability of active thermotolerant oil-oxidizing microorganisms to degrade oil at different concentrations of mineral salts (Cl-, SO42-) and to determine the emulsifying activity to assess theirbiotechnological potential.It was established that all selected strains are halotolerant. Moreover, the temperature factor influenced the degree of oil degradation in the presence of mineral salts. The highest percentage of oildegradation was at 35°C. At this temperature, strains K-3, 4/5, P1-35-14 showed high activity at allstudied NaCl concentrations (3, 5 and 10%) – 73.6-85.9%, 62.7-91.1 % and 73.6-75.7%, respectively.At 40°C, the strain IP-40-4 showed the highest activity – 41.4-51.2%. At 50°C, the destruction abilityof the studied cultures was approximately at the same level at all NaCl concentrations. The cultivationof selected strains with a high content of MgSO4 in the medium showed that the K-3 strain was the moststable, which remained highly active at all temperatures.It was shown that active strains of thermotolerant oil-oxidizing microorganisms had emulsifying activity. High values of emulsifying activity are observed at a temperature of 35°C, and with a subsequentincrease in temperature, the formation of surfactants decreased. At 35°C, strain P2-35-9 had the greatestemulsifying activity, and at 40°C and 50°C – strain K-3.Key words: bioremediation, thermotolerant hydrocarbon-oxidizing bacteria, salinity, halotolerance,emulsifying activity.

Tissue damage to wheat seedlings (<i>Triticum aestivum</i>) under salt exposure

High content of mineral salts reduces crop yields. Agricultural areas are being reduced due to the secondary salinization of land. Wheat is one of the main crops, however, wheat is a type of plant that is sensitive to the toxic effects of mineral salts. Therefore, the identification of cytological and biochemical markers for determining resistant and sensitive wheat varieties to abiotic stress caused by the toxic effects of mineral salts is one of the important tasks of agriculture. Methods of fluorescence, cytophotometry, electrophoresis, and light-optical microscopy produced suitable cytological markers for creating a test system for the early diagnosis of wheat sensitivity to the tested salts. Different wheat genotypes of Triticum aestivum Host. (varieties Agata, Uchitel’, and Zhemchuzhina Povolzh’ya) underwent treatment to investigate their reaction to the toxic effects of sodium chloride and sodium sulfate. The varieties responded differently to the presence of the salts; these were reflected in changes in biometric data, the passage of the cell cycle, features of ROS accumulation, and a quantitative assessment of the viability of seedling cells. In the genotypes sensitive to sodium chloride and sodium sulfate (varieties Zhemchuzhina Povolzh’ya and Uchitel’), tissue damage was observed in root cells under toxic effects, while in the resistant genotype (variety Agata) cell damage was minimal. The developed complex of intravital markers of various wheat genotypes in the early development stages can be effectively used to characterize wheat with different tolerance to chloride and sulfate salinization.

Assessment of Usability of Thermal Bicarbonate Mineral Water from the “Odra-5/I” Intake in Grabin Opole Silesia (SW Poland) for Therapeutic and Prophylactic Purposes

Thermal bicarbonate mineral water belong to sought-after balneological resources. This kind of water are a balneological product and valuable raw material used in health resorts for the wider health strategy - prophylaxis and recreation. The discovery of thermal bicarbonate mineral water in Grabin near Opole in 1983 was purely incidental when a intake was made for the recognition of the geological structure of the crystalline basement of Mesozoic and Cenozoic sediments. Thermal bicarbonate mineral water existing in Grabin can be found in the Fore-Sudetic block and is a unique phenomenon. Its chemical composition, and especially high total mineral salt content of the waters from Grabin, indicate a type of mineral water unparalleled in Poland. The content of solids is much higher than in all other waters existing in e.g. Lower Silesian health resorts. Its special feature is also the source performance, which is 150-200m3/h and is unprecedented when compared to other sources in the region. The discovery of thermal bicarbonate mineral water in Grabin also puts a new light on the presence of perspective areas for drawing waters useful in medical care and prevention.

On the Validation of Satellite Microwave Remote Sensing Data under Soil Salinity Conditions

The results of a comparative analysis of the underlying surface brightness temperature obtained from the SMOS satellite and calculated with the use of ground-based measurements of moisture and the laboratory measurements of dielectric characteristics of samples of saline soils and saline water from test sites of the Kulunda Steppe are presented. The values of the underlying surface brightness temperatures calculated with the use of ground investigations and the laboratory studies correlate satisfactorily with the values of brightness temperatures measured from the SMOS satellite. The deviations can be caused by the seasonal drying of highly mineralized shallow lakes, which results in a significant change in the areas occupied by lakes and the appearance (instead of a water surface) of wetlands with a high content of mineral salts, whose radio emission characteristics differ from the characteristics of the water surface and the soil cover.

Use of Sieving As a Valuable Technology to Produce Enriched Buckwheat Flours: A Preliminary Study.

Fractionation processes based on physical separation are a good strategy to produce enriched cereal flours. Therefore, the aim of this work is to evaluate the suitability of sieving of buckwheat flours to produce protein and phenolic (especially rutin) enriched fractions. Because of that, dehulled whole buckwheat flour (GSTQ) was sieved obtaining fractions with a particle size of 215 µm, 160 µm, 85 µm, and 45 µm (GS215, GS160, GS85, and GS45). For that purpose, the determination of protein, ash, and total starch content and free and bound phenolic compounds was carried out. The highest content of total phenolic compounds was obtained in GS215 (3118.84 mg kg−1 d.w.), followed by GS160 (2499.11 mg kg−1 d.w.), GS85 (989.46 mg kg−1 d.w.), GSTQ (983.15 mg kg−1 d.w.), and GS45 (481.31 mg kg−1 d.w.). Therefore, the phenolic content decreased with the particle size decrease from 215 µm to 45 µm. Besides, there were no significant differences between the total phenolic content in GS85 and GSTQ. The fraction with 215 µm reported the highest protein and mineral salt content and presented rutin amounts four times higher than GSTQ.

Degradation of complex casein polymer: Production and optimization of a novel serine metalloprotease from Aspergillus niger KIBGE-IB36

Abstract Serine metalloproteases have substantial scientific interest due to their unique physicochemical properties and capability to degrade complex casein polymer. In this contemporary study, the production of protease was enhanced by optimizing various physicochemical parameters and one-variable at a time approach used under submerged fermentation conditions. Among four different previously isolated Aspergillus species, Aspergillus niger KIBGE-IB36 represented high protease titer along with casein hydrolyzing zone on casein agar plate. Maximum protease production was achieved at 30 °C after 5 days of cultivation period at pH-6.0. Glucose (2.5 g L-1) and casein (2.5 g L-1) augmented protease production within the growth medium. Further increment in protease production was attained when different combinations of nitrogen and mineral salts concentrations used. Peptone (20.0 g L-1), yeast extract (0.50 g L-1), CaCl2 (0.10 g L-1), MgSO4 (0.10 g L-1) and K2HPO4 (0.30 g L-1) boosted the protease secretion. The current study is imperative for the large scale production of serine metalloprotease that also possess extraordinary prominence in food processing industries.

DEVELOPMENT OF A PROCESS CONTROL SYSTEM FOR DYNAMIC SEDIMENT OF OIL EMULSION


 
 
 Using heat balance and geometric features of horizontal cylindrical sedimentation, a new method and algorithm for controlling the dynamic sediment of emulsified water drops of oil emulsion have been developed. The mechanism of oil emulsion dynamic sediment on the proposed method is that the redistribution of the flow contributes to the cyclic change in the flow rate of the oil emulsion in the settling apparatus and the oscillatory motion (compression and expansion) of the intermediate emulsion layers, leading to the destruction of armoring casings, coalescence of emulsified water drops and transfer mechanical impurities into water cushion of the settling apparatus, as a result of which the quality of commercial oil increases (the content of water and mineral salts decrease in the prepared oil) and the risk of flooding the settling apparatus decreases. In the settling apparatus, where the volume of the oil emulsion is greater than the average value, the intermediate emulsion layer expands, the kinetic energy increases, and it increases the efficiency of collisions between the drops, leading to the destruction of the armoring casings and coalescence of the drops. When the volume is less than the average value, the intermediate emulsion layer is compressed, the distance between the drops decreases, leading to a coalescence of the drops and an increase in the efficiency of oil preparation.
 
 

Removal of metronidazole antibiotic pharmaceutical from aqueous solution using TiO2/Fe2O3/GO photocatalyst: Experimental study on the effects of mineral salts

A TiO2/Fe2O3/GO photocatalyst is synthesized via the sol-gel method and characterized by X-Ray Diffraction (XRD), X-Ray Fluorescence (XRF), FT-IR, Brunauer-Emmett-Teller (BET), and Ultraviolet-Visible Diffuse Reflection Spectroscopy (UV-Vis DRS) analyses. Metronidazole (MET) concentration (10-20 mg/L), photocatalyst concentration (0.5-1.5 g/L), irradiation time (60-120 min), and initial pH (4-6) are investigated through response surface methodology (RSM), and the optimal process conditions are determined. The removal efficiency of MET with the TiO2/Fe2O3/GO photocatalyst is 97% under optimal conditions: a pollutant concentration of 10 mg/L, the irradiation time of 120 min, photocatalyst concentration of 1 g/L, and pH of 5. The influence of mineral salts concentrations (50-800 mg/L), including NaCl, Na2SO4, NaHCO3, KCl, MgSO4, and CaCl2, are examined at the initial pH of 5, photocatalyst concentration of 1 g/L, and pollutant concentration of 20 mg/L. According to the results, the reaction rate constant decreases with an increase in mineral salts concentrations up to 800 mg/L, especially with Na2SO4 (42.43% deactivation) and also with MgSO4 (38.08%) and NaHCO3 (37.73%), under the same operational conditions. The effects of mineral salts such as NaCl and KCl on the reaction rate constant for the contaminant removal efficiency have a downward trend until these salts reach a 200 mg/L concentration, and then they experience an upward trend.

MINERAL SALT CONTENT IN THE WATER OF THE VOGHJI RIVER CATCHMENT BASIN (ARMENIA): ASSESSMENT OF POTENTIAL EFFECTS ON IRRIGATED SOILS AND CROPS

Agricultural risks of mineral salt (Ca2+, Mg2+, K+, Na+, SO42- , Cl- , CO32- , HCO3- ) content in the water of the Voghji River catchment basin were assessed. Water sampling and hydrochemical analyses were implemented in July and September 2017. The results of the study showed that anthropogenic salt level in the Voghji River water site located downstream from Kapan Town and in the area of the Norashenik River mouth may have posed serious health risks to irrigated soils and crops.

Mineral salt intake effects on faecal-N concentration and the volume and composition of beef cattle urine.

The effect of mineral salts on water ingestion and urine volume in cattle has been extensively studied. However, recently, this effect has been investigated as a potential mitigator of environmental aspects related to the nitrogen (N) cycle, such as nitrate (NO3-) lixiviation, ammonia (NH3) volatilisation, and nitrous oxide (N2O) emissions. The effect of mineral salts, particularly sodium chloride (NaCl), on urine-N concentration, urine volume, the proportion of N compounds in the urine, and faecal-N concentration has not yet been explored in field conditions with respect to environmental aspects of beef cattle production. The present study investigated the effect of dietary mineral salt rates on these parameters. A Latin square (5 × 5) experimental design was utilised with five concentrations of mineral salts in the diet: 0.0, 2.0, 4.0, 6.0, and 8.0g based on dry matter (DM) ingestion (g/kg DM). The nitrogen concentration in the urine and urine volume increased linearly. The total N excreted (g/day) via urine did not vary with increasing mineral salt concentrations. When evaluated, the N compounds of urine (urea-N, allantoin-N, and hippuric acid-N) also reacted to the increased mineral salt concentrations, while creatinine-N did not. Urea-N, allantoin-N, and hippuric acid-N linearly increased their proportions in total N-urine. The N concentration in faeces was not affected by mineral salt concentrations. The urine volume, concentration of N, and proportion of N compounds in the urine affected N2O emissions and NH3 volatilisation. Therefore, mineral salt utilisation may be an option for mitigating N pollution from beef cattle, especially for grasslands in tropical countries.

Uranium concentrations in sediment pore waters of Lake Neusiedl, Austria

The goal of the present investigation was to measure 234U/238U activity ratios in pore waters of Lake Neusiedl, Austria, in order to learn more about uranium in groundwaters of the Lake Neusiedl/Seewinkel region. Pore waters of waterlogged sediments (at 1m depth) in the littoral zone of Lake Neusiedl were analyzed. The pore water samples were collected in the National Park Lake Neusiedl/Seewinkel from pristine sites that were not influenced by neighboring fertilized fields or vineyards. Uranium isotopes were extracted from 1.5L of sediment pore water and measured by α-particle spectrometry. Uranium concentrations were found to be unexpectedly high (up to 853μgL−1) especially in pore waters of salt-rich locations. 234U/238U activity ratios were between 0.91 and 1.09 for all pore water samples, irrespective of their origin from the east or west littoral zones of the lake. Uranium and mineral salts concentrations were strongly correlated. 222Rn concentrations were low (between 22 and 42BqL−1). The results provide insight into the high degree of mobility of U(VI) in sedimentary environments, in the presence of migrating Na2SO4-type saline waters.

In vitro cultivation of purple basil Ocimum basilicum L. ‘red rubin’ at different levels of salts, charcoal, sucrose and potassium iodine

Ocimum basilicum'red rubin' is highly valued for its essential oil, and is widely used in folk medicine, but few studies examining in vitro propagation techniques for this basil variety are available. We examined the development of red rubin basil seedlings grown in vitro in different concentrations of mineral salts, charcoal combined with sucrose, and potassium iodine in three series of tests. In the first series, five concentrations (0, 25, 50, 75 and 100%) of Murashige and Skoog salts (MS culture medium) were used to germinate sterilized seeds of O. basilicum. In the second test, to evaluate the combined effects of charcoal and sucrose on basil growth, three concentrations of charcoal (0, 3.0, and 4.5 g/L) and two of sucrose (30 and 60 g/L) were used. In the third test, the seeds were sown onto culture medium enriched with five different concentrations of potassium iodine (0, 25, 50, 75 and 100 M); 0.5 M benzyl aminopurine (BAP) and 1.0 M naphthalene acetic acid (NAA) were added to all germination media. A completely randomized experimental design was used in all tests, with five replicates, four vials per test, and 4 seeds per vial. In Test 2, seedling oxidation (%) and the numbers of leaves and shoots were submitted to non-parametric analysis using the Kruskal Wallis test (p =0.05). The other data were submitted to analysis of variance (p=0.05). The qualitative means were evaluated by the Tukey test (p=0.05), and the quantitative means by polynomial regression (p=0.05). The results showed that 100% MS culture medium was most indicated for 'red rubin' seed germination and aerial portion production, while 70-80% MS was indicated for root production. A sucrose level of 30 g/L (standard) combined with 4.5 g/L of activated charcoal allowed the best seedlings development and inhibited callus formation. Activated charcoal limited copper and zinc uptake by O. basilicum seedlings, but not enough to hamper their development. Increasing concentrations of potassium iodine reduced all of the growth variables analyzed in this basil variety.

A consortium of immobilized rhodococci for oilfield wastewater treatment in a column bioreactor

The possible use of a consortium of actinobacteria from the genus Rhodococcus immobilized on a polymeric carrier has been investigated for oilfield wastewater treatment in a bioreactor. It has been found that Rhodococcus opacus IEGM 263 and Rhodococcus ruber IEGM 231 cells remain viable at high concentrations of mineral salts in water and are able to oxidize oil hydrocarbons up to 62–81%. It has been shown that the consortium of rhodococci was more efficient in the elimination of hydrocarbons from wastewater than monocultures.

<b>Developing an <i>in vitro</i> optimized protocol to sweet potato landraces conservation

This study aimed to develop a protocol for the in vitro conservation of 30 accessions of sweet potato collected in the northern region of Rio de Janeiro State, Brazil, aiming to extend the period between subcultures for this species. An in vitro minimal growth experiment was established with accession UENF 1931 in a factorial arrangement consisting of four concentrations of mineral salts in MS medium and four concentrations of sucrose, at two temperatures. The others accessions were grown in medium containing 100% MS salts and 2% sucrose, which was the medium that allowed the in vitro conservation of accession UENF 1931 for a longer time period. For this second experiment, 30 accessions of sweet potato and two temperatures were used. Plant height, number of leaves, and survival rate were measured every 30 days for 12 months, in both experiments. In the last stage, the sweet potato plantlets originating from in vitro minimal growth were acclimatized and exhibited 100% survival. The recommended in vitro minimal growth conditions for the studied accessions are MS medium with 100% concentration of mineral salts and 2% sucrose, at a temperature of 27±2°C, with subculture performed every 180 days.

The influence of humic acids on desalination process with the use of electrodialysis

With every year amount of drinking water in the world becomes smaller. One of the possible way to solve the problem with water scarcity is desalination. The aim of this paper is to evaluate the use of electrodialysis for salt removal in the presence of organic matter. During the tests installation PCCell BED-1-System was applied. Desalination was conducted with the use of standard ion-exchange membranes. The treated solutions contained 0.5 and 1 g NaCl/dm3. The solution poured into a diluate chamber was also supplemented with humic acids (concentration amounted to 5, 10 and 15 mg/dm3). The current intensity was equal to 0.11 A (current density equal to 1.72 mA/cm2). The process was terminated when voltage reached 24 V. During the tests conductivity, colour and voltage was monitored. Also specific electrical energy demand (EC) was calculated. It has been found that electrodialysis can be used as an effective method for salt removal. In most cases conductivity was reduced by approximately 90%. The main factor which influenced EC and process duration was mineral salt content. Besides, it has been observed that organic matter had a slight impact on the process course.

De novo assembly and analysis of changes in the protein-coding transcriptome of the freshwater shrimp Paratya australiensis (Decapoda: Atyidae) in response to acid sulfate drainage water.

BackgroundThe atyid shrimp Paratya australiensis occurs in surface freshwater habitats throughout eastern Australia and has been used to study the ecotoxicology of contaminants such as pesticides and metals. The acidification of surface water that can occur after acid sulfate material in soils and sediments is oxidised and subsequently re-wetted is a serious environmental issue in coastal regions and inland riverine floodplains worldwide. Solubilisation of soil-associated minerals can result in high waterborne concentrations of mineral salts and dissolved metals, which together with low pH represent a potential threat to aquatic ecosystems in affected regions. The aims of the present study were to gain insight into stress responses induced by exposure to acid drainage water (ADW) in P. australiensis by determining changes in the abundance of protein-coding transcripts and to generate a comprehensive transcriptomic resource to facilitate further research into gene regulation or protein structure and function in this species. Adult P. australiensis were exposed for 24 h to undiluted ADW, 50 % ADW diluted in river water, or to river water as control, and high-throughput mRNA sequencing (RNA-Seq) conducted on whole-body tissues. A reference transcriptome was generated using de novo assembly and putative protein-coding regions were identified and annotated. Changes in transcript abundance in response to ADW exposure were determined by aligning reads to the reference transcriptome and quantifying coverage.ResultsA high proportion of arthropod benchmarking universal single-copy orthologues were present in the reference transcriptome. Functions associated with cuticle biosynthesis and oxidative stress were significantly enriched in the lists of transcripts exhibiting differential abundance in either direction after exposure to 50 % or 100 % ADW. Transcripts involved in osmoregulation exhibited decreased abundance following exposure to ADW. The transcriptome contained full-length coding sequences for numerous proteins known to be involved in environmental response pathways, including two putative metallothioneins, four glutathione peroxidases and 19 nuclear receptors.ConclusionsThe results of the present study provide insight into stress response pathways induced in crustaceans by short-term exposure to multiple stressors present in ADW such as low pH, high salinity and dissolved metals, and represent a resource for future toxicogenomics and protein functional studies in P. australiensis.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3208-y) contains supplementary material, which is available to authorized users.

In vitro propagation of the endangered tree fern Cibotium barometz through formation of green globular bodies

Cibotium barometz is an endangered tree fern, used both as ornamental plant and traditional Chinese medicinal plant. In this study, an effective in vitro propagation protocol was obtained through formation of green globular bodies (GGBs) from in vitro juvenile sporophytes. The effect of plant growth regulators (PGRs) on GGB induction and multiplication, as well as mineral salt concentration and active charcoal (AC) on plantlet regeneration from GGBs was evaluated. Thidiazuron (TDZ; 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea) had a significant effect on GGB induction and multiplication (P 0.05). GGB induction rate was above 80 % on 1/2 Murashige and Skoog (MS) media supplemented with TDZ (1.0 mg L− 1) and NAA (0.1, 0.3 or 0.5 mg L− 1). The same media were also optimal for GGB multiplication. GGBs cultured on 1/4 MS media supplemented with 0.1 or 0.2 % (w/v) AC showed a high rate of GGB development into plantlets above 90 %. 1/2 MS media supplemented with 0.1 or 0.2 % AC were the most effective for plantlet growth. Regenerated plantlets were successfully acclimatized (80 %) in greenhouse conditions. Morphological and histological analysis revealed that C. barometz GGBs was a yellow-green globular structure composed of the single GGB with meristems and hair-like structures, and new single GGBs were initiated from the epidermal cells of meristem zone.

Suitability of Beer As An Alternative to Classical Fitness Drinks

During longer physical strains, our body loses water and salts through sweating. This deprivation leads to states of exhaustion and convulsions. A fitness drink should replenish both, water and mineral nutrients (mainly sodium). Different beers, alcoholic, alcohol-free and yeast-clouded, alcohol-free beer were analyzed by HPLC and Ion chromatography to determine the content of mineral salts and carbohydrates. Osmolality as a degree of tonicity was calculated based on the dissolved components. Data was compared to fitness drinks declared as mineral nutrient containing and isotonic. Subsequently, different sodium salts were added to alcohol-free, yeast-clouded beer to reach the EU recommended sodium concentrations of 500 mg L-1. These spiked beers were blind tested for flavor impairments. This study shows that the fitness drinks came close to fulfill the EU direction. Generally, almost all beers are mineral nutrient containing, as defined by Austrian law. Due to the alcohol concentration, none of alcoholic beers are isotonic, as they are hypertonic. Most of the alcohol free beers are within the isotonic range, especially considering dissolved carbon dioxide. After ingestion, it is degassed by the acidic conditions in the stomach, changing many beers to hypotonic in the end. Each beer fulfills the EU recommended carbohydrate content but none comes even close to reaching the required sodium content. The blind tasting of the spiked beers showed minor flavor impairment with sodium carbonate causing the least negative effects on taste and odor. In conclusion, yeast clouded, alcohol-free beer might be labeled as mineral nutrient containing and isotonic, implying to be an ideal sport drink. Due to the lack of sodium, the real benefit of such a beverage could diminish unless mixed with beverages containing high levels of sodium or sodium carbonate.

Integration of tissue culture and cryopreservation methods for propagation and conservation of the fern Osmunda regalis L.

A simple and reliable technique for in vitro multiplication and long-term preservation using liquid nitrogen was developed for gametophytes of Osmunda regalis. The effect of Knop’s medium and various concentrations (1/2, 1/4, 1/8) of mineral salts provided in Murashige and Skoog (MS) basal medium, together in the presence or absence of both ammonium nitrate and a full complement of vitamins on gametophyte proliferation and sporophyte production was determined. Moreover, the effectiveness of gametophyte cryopreservation by vitrification and encapsulation–vitrification techniques was assessed. Maximum gametophyte proliferation (89 %) occurred on the ammonium nitrate- and vitamin-free [(-NH4NO3)(-vit)] MS medium with 1/2 or 1/4 strength mineral salts. The maximum production of sporophytes (30 plantlets per gametophyte clump) required 1/8MS (-NH4NO3)(-vit) medium. The flow cytometric analysis revealed that the sporophytes contained twofold more pg DNA than gametophytes. This confirmed that the sporophytes were obtained by sexual reproduction. The vitrification protocol and PVS2 solution were ineffective for cryopreservation. The greatest survival rate (81.6 %) following cryo-exposure occurred following treatment of encapsulated gametophytes with PVS3 solution for 3 h. This protocol allowed the recovery of gametophyte cultures following 6 weeks after rewarming. Finally, 100 % of sporophytes produced in vitro were successfully acclimated to ex vitro conditions. Application of the in vitro and cryopreservation methods made it possible to improve the number and time of O. regalis sporophyte production. Whole system of micropropagation can be completed in approximately 1 year. The protocols open new avenues for the mass propagation, germplasm conservation and resource management of the species.