Rehydration Media Research Articles

Вплив трифторперазину та децилсульфату натрію на осмотичний шок еритроцитів людини та кролика

The effects of trifluoroperazine and sodium decyl sulfate on posthypertonic shock of human and rabbit erythrocytes were studied. For this purpose, the level of hemolysis in posthypertonic shock and the percentage of potassium ions released from erythrocytes in dehydration and rehydration solutions in the presence of trifluoroperazine and sodium decyl sulfate were determined. It is shown that the protective effect of amphiphilic compounds is manifested at 0°C, but not at 37°C. There is a decrease in the level of hemolysis in a certain concentration range of each of the studied substances. It was found that human erythrocytes are more sensitive to the protective action of the studied amphiphilic compounds and are characterized by a wider range of protective concentrations. This could be explained by the different lipid composition of the erythrocyte membranes of the studied mammals. Measurement of the leak of potassium ions in dehydration and rehydration media in the presence of amphiphilic compounds suggested that the latter do not affect the permeability of the membrane of mammalian erythrocytes for potassium in posthypertonic shock. Based on the obtained results and literature data, it is assumed that the protective effects of trifluoroperazine and sodium decyl sulfate occur independenly of the formation of temporary defects in permeability for potassium ions, but involve an increase in the cell surface area due to the incorporation of amphiphilic molecules. This results to an increase in the critical hemolytic volume of erythrocytes and reduction in the level of damage during change from hypertonic conditions to isotonic ones.

Inorganic Additive Improves the Survival of the Probiotic Lacticaseibacillus rhamnosus CRL1505 During Spray Drying, Rehydration, and Storage.

In previous in vitro studies, an inorganic additive (MCM3) showed a thermo-protective effect on the cell viability of Lacticaseibacillus rhamnosus CRL1505 (Lr-CRL1505). In this work, cultures of this probiotic strain were spray dried at lab scale using two carriers: maltodextrin (powder MA) and maltodextrin plus MCM3 (powder MA/MCM3). The cell survival was higher in powder MA/MCM3 (72.8%) than in powder MA (42.8%). Different rehydration media, including the additive MCM3, and two temperatures (37°C and 45°C) were evaluated. The best results were obtained in cells rehydrated at 37°C in MCM3. During the storage of the powders, the highest cell counts were observed in the MA/MCM3 powder. Our results demonstrated that the presence of MCM3 in the carrier and in reconstitution media benefits the spray drying process and the recovery of dehydrated cells. Thus, the use of this additive of inorganic nature and low cost represents a promising technological alternative.

Effect of lyophilization on food grade liposomes loaded with conjugated linoleic acid

The effect of lyophilization and rehydration medium on a liposome system for conjugated linoleic acid (CLA) delivery was studied. Liposomes were prepared by ethanol injection method employing soy phosphatidylcholine and CLA isomers 9c, 11t and 10t, 12c. Two rehydration media were assessed: a food simulant (distilled water) and the original medium used for liposome preparation (ethanol: water 1:9). Rehydrated liposomes were characterized in size, morphology, CLA encapsulation efficiency and membrane fluidity, at 3 and 30 days of cold storage. CLA loaded liposomes presented better stability and lower size than control liposomes without CLA through storage time, regardless of the rehydration medium. These facts could be related with spin label EPR measurements: CLA disordered the outer part of the lipid bilayer increasing fluidity, and ordered the inner part of the bilayer decreasing fluidity, which indicates a more packed membrane in the hydrophobic region. In all cases Transmission Electronic Microscopy images showed nanometric sized and oligo-lamellar vesicles. Liposomes preserved CLA isomers with high encapsulation efficiency: no differences were noticed between fresh and lyophilized samples. No influence of rehydration medium was noticed for the majority of the studied parameters. We have successfully developed efficient liposomal systems for bioactive compounds delivery in food applications.

Impact of the CaCl2 content in the rehydration media on the microcapsule formation out of spray dried capsule precursors for the immobilization of probiotic bacteria

The aim of this work was to develop a capsule formation process at moderate temperatures with a high encapsulation yield (EY) of probiotic bacteria. Capsule precursors (CP) obtained by spray drying were rehydrated at 40 °C in aqueous solution to form water-insoluble hydrogel capsules. The CaCl2 concentration (0–2 mm) in the rehydration solution was varied. The rehydration step was evaluated regarding matrix stability and capsule size. The results show that only low amounts of non-aggregated casein remain in the rehydration media at or above 1.3 mm CaCl2. Additionally, under this condition the particle size distribution shows no particles with the size of non-aggregated caseins and bacteria. Both results indicate a completely, stable matrix structure. To gain a deeper understanding the EY and capsule morphology by scanning electron microscope (SEM) images were assessed. The SEM images show that the bacterial cells are fully embedded in a dense casein network structure. The determined EY was ∼85%.

The influence of various rehydration protocols on biomechanical properties of different acellular tissue matrices.

This study evaluated the influence of different rehydration media and time periods on biomechanical and structural properties of different acellular collagen matrices (ACMs). Specimens of three ACMs (mucoderm®, Mucograft®, Dynamatrix®) were rehydrated in saline solution (SS) or human blood for different time periods (5-60min). ACMs under dry condition served as controls. Biomechanical properties of the ACMs after different rehydration periods were determined by means of tensile testing. ACMs' properties were further characterized using Fourier-transform-infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). At dry conditions, mucoderm® presented the highest tensile strength (TS) and Dynamatrix® showed the maximum elastic modulus (EM; p each ≤0.036). Rehydration in SS and blood resulted in significant TS changes of mucoderm® (p each ≤0.05). Concering EM, mucograft® showed significantly decreased values after rehydration in SS compared to Dynamatrix® and mucoderm® after 10min (p each ≤0.024). mucoderm® hydrated for 5min in blood displayed nearly double TS and a significantly increased EM after 60min (p = 0.043) compared to rehydration in SS. TS and EM values of Dynamatrix® and Mucograft® were not altered following rehydration in blood versus SS (p each ≥0.053). FTIR analysis confirmed the recovery of the graft protein backbone with increased rehydration in all samples. DSC measurements revealed that tissue hydration decreased thermal stability of the investigated ACMs. Our findings demonstrated that the rehydration protocol affects the biomechanical properties of ACMs. Clinicians should be aware of altered handling and mechanical properties of ACMs following different rehydration protocols.

Effect of growth phase, protective agents, rehydration media and stress pretreatments on viability of Oenococcus oeni subjected to freeze-drying

Malolactic fermentation (MLF) is an important process in wine production. To achieve successful MLF, expanding interest in ready-to-use Oenococcus oeni starter cultures has drawn greater emphasis on developing starter production and preservation methods. Growth phase, protective agents, rehydration media and stress pretreatments were the main factors to affect the survival of O. oeni SD-2a after freeze-drying. It was found thatO. oeni SD-2a cells in the early stationary phase survived better than those in the mid-log phase after freeze-drying. Protective agents play an important role in the conservation of viability. Sodium glutamate (2.5%) was the best protectant, giving the cell viability 69.5%. The addition of polysaccharides and disaccharides in suspension media also improved the cell viability signii¬cantly. Rehydration is a critical step in recovery after freeze-drying. When freeze-dried O. oeni SD-2a was rehydrated in acidic tomato broth (ATB) medium, the highest viability was obtained. Rehydration in the tested disaccharide solutions decreased the cell viability obviously. During stress treatments, O. oeni SD-2a treated with 8% ethanol resulted in the highest freeze-drying survival rate (82%), the treatment with pH 3.5 or 3.2 also notably increased freeze-drying viability. Key words: Oenococcus oeni, malolactic fermentation and freeze-drying and viability.

Freezing and freeze-drying of the bacterium Rahnella aquatilis BNM 0523: study of protecting agents, rehydration media and freezing temperatures

The aim of the present study was to evaluate and compare freezing and freeze-drying treatments for conserving Rahnella aquatilis (BNM 0523) with the goal to achieve an adequate commercial formulation of this biocontrol agent. The effect of several protective agents, rehydration media and freezing temperatures on the viability and functional activity of the R. aquatilis was investigated. The storage stability at 3 months and 4 years was determined by checking the viability of the cells and their biocontrol capability against Botrytis cinerea by measuring the percentage of reduction of disease severity on apple. The best results were obtained by the freeze-drying of the cells using a mixture of skimmed nonfat milk 10%, yeast extract 0·5% and glucose 1% as the protecting and rehydrating medium, and a quickly freezing (-70°C) before the freeze-drying. In this case, the viability of the cells after 4 years was 98%, and their antagonistic ability showed a little decrease with respect fresh cells. The studies showed that R. aquatilis was resistant to freezing and freeze-drying when it was used a mixture of cryoprotectants and that it was possible to obtain inoculums with high viability and good effectiveness for reduction of decay caused by B. cinerea. This study is probably the first report about the resistance of R. aquatilis to freezing and freeze-drying treatments and shows that these operations could be useful for obtaining a commercial formulation of this biocontrol agent.

Quantitative expression analysis of mleP gene and two genes involved in the ABC transport system in Oenococcus oeni during rehydration

Oenococcus oeni is recognized as the principal microorganism responsible for malolactic fermentation, and the control of its activity is of primary importance in winemaking. The aim of this study was to quantify the levels of expression of the malate transporter gene (mleP) and of two genes putatively involved in the ATP-binding cassette transport system (oeoe_1651, oeoe_0550) to better understand the physiological response of bacteria during rehydration. These genes coding for transporters were studied in different rehydration media. Initially, three different statistical algorithms were used to identify suitable reference genes to be used for the normalization of expression data in O. oeni during rehydration, and to this purpose, the best genes found were ddl and gyrB. The results showed that the genes for transporters of malate and sugar (mleP, oeoe_1651) were activated immediately after a few minutes of rehydration, when specific medium compositions were used.

Characterization of high-milk-protein powders upon rehydration under various salt concentrations

Rehydration of native micellar casein and native whey isolate protein powders was followed in different ionic environments. Solutions of NaCl and CaCl2 in the concentration range of 0 to 12% (wt%) were used as rehydration media. The rehydration profiles obtained were interpreted in terms of wetting, swelling, and dispersion stages by using a turbidity method. Two behaviors were observed depending on the salt concentration. For native micellar casein powder, a significant change was observed between 3 and 6% NaCl and between 0.75 and 1.5% CaCl2. The first behavior (low salt concentration) presents a typical rehydration profile: quick wetting, swelling, and long dispersion stage. The dispersion stage of the second behavior (high salt concentration) was significantly shortened, indicating a strong modification of the protein backbone. The rehydration of whey protein powder was less influenced by salts. At low salt concentrations, a typical profile for whey powders was observed: wetting with lump formation and no swelling followed by a quick dispersion. At high CaCl2 concentrations, no turbidity stabilization was observed, indicating a possible protein unfolding and denaturation. Additionally, the changes in secondary structures of the 2 proteins upon salt increase were followed by Fourier transform infrared spectroscopy and confirmed the different profiles observed.

Influences of protectants, rehydration media and storage on the viability of freeze-dried Oenococcus oeni for malolactic fermentation

Malolactic fermentation (MLF) is an important process in wine production. To achieve successful MLF, expanding interest in ready-to-use Oenococcus oeni starter cultures has placed greater emphasis on developing starter production and preservation methods. In this study, influences of protectants, rehydration media and storage on the viability of O. oeni H-5 when subjected to freeze-drying were investigated. It was found that sodium glutamate (2.5%) was the best protectant, giving the cell viability 72.4%. Adding polysaccharides and disaccharides in suspension media also improved significantly the cell viability. Rehydration is an important step in recovery after freeze-drying. When freeze-dried O. oeni was rehydrated in GYM medium, the highest viability (87.1%) was obtained. Rehydration in the disaccharide solutions tested made the cell viability obviously decrease. After 6 months storage at 4°C, loss of viability occurred, the extent of which depended on protectants used, sodium glutamate again being the most effective.

Survival of the postharvest biocontrol yeast Candida sake CPA-1 after dehydration by spray-drying

Spray drying was evaluated as a dehydrating method to preserve the postharvest biocontrol agent Candida sake CPA-1. The effect of drying temperature, carrier, growth and rehydrating medium on the survival of the yeast was studied. Outlet temperature had more influence on the death of the cells than inlet temperature, and survival decreased with increasing temperature. Spray drying at an inlet temperature of 150°C was optimum in terms of viability, powder recovery and moisture content of the product. Use of 10% (v/v) skimmed milk as a carrier gave the highest survival and percentage of powder recovery (34–47%). Rich rehydration media were found to be better than water or phosphate buffer, with slight differences on survival. Spray-dried cells were less effective than fresh ones in controlling Penicillium expansum rot on apples. Spray drying of C. sake was not a good dehydration method as it gave low cell survival, poor recovery of product, and low efficacy.

Effect of protective agents, freezing temperature, rehydration media on viability of malolactic bacteria subjected to freeze-drying

The effects of protective agents, rehydration media and freezing temperature on the viabilities of Lactobacillus brevis and Oenococcus oeni H-2 when subjected to freeze-drying were investigated. Several protectants and rehydration media were tested to improve the survival after freeze-drying. The cells were also frozen at -65 and -20 degrees C to check the effect of freezing temperature on the viability. The best protectant and rehydration medium to obtain the highest viability after freeze-drying varied with the species of bacteria. Yeast extract (4.0%) and sodium glutamate (2.5% ) gave maximum viability of L. brevis and O. oeni (67.8% and 53.6% respectively). The highest survival of L. brevis and O. oeni were obtained when rehydrated with 10% sucrose and MGY medium respectively. When the bacterial cells were frozen quickly (-65 degrees C) than slowly (-20 degrees C), L. brevis and O. oeni both showed increased viability after freeze-drying. The viabilities of L. brevis and O. oeni after freeze-drying were shown to be strain specific and dependent on protective agents, rehydration media and freezing temperature.

Viability, Efficacy, and Storage Stability of Freeze-Dried Biocontrol Agent Candida sake Using Different Protective and Rehydration Media

Viability, efficacy against Penicillium expansum on Golden Delicious apples, and storage stability of freeze-dried Candida sake strain CPA-1 were studied. The effect of several protective agents and rehydration media was investigated in the freeze drying of C. sake. Skimmed milk at 10% concentration was a good rehydration medium for all protectants tested. In general, good viability results were obtained when the same solution was used as a protectant and as a rehydration medium. The best survival was obtained when C. sake cells were protected with 10% lactose + 10% skimmed milk and rehydrated with skimmed milk (85% viability). The potential for biocontrol of the best freeze-dried treatments against P. expansum on apples was compared with that of fresh cells. Freeze-dried treatments at 1 × 107 CFU/ml reduced the incidence of decay by 45 to 66%. The best biocontrol effect was obtained with cells that had been freeze dried using 10% lactose + 10% skimmed milk as a protectant and 1% peptone as a rehydration medium, with a 66% reduction in rot incidence. However, in all treatments, the efficacy of freeze-dried cells was significantly lower than fresh cells. The stability of freeze-dried samples decreased during storage and was influenced by storage temperature. In the best treatment, storage of C. sake cells for 60 days at 4°C resulted in final concentrations of 2.5 × 108 CFU/ml, which was a 10-fold reduction in relation to the initial starting concentration of cells prior to freeze drying.

Effect of protective agents, rehydration media and initial cell concentration on viability of Pantoea agglomerans strain CPA-2 subjected to freeze-drying

The effect of initial cell density, protective agents and rehydration media on the viability of biocontrol agent Pantoea agglomerans CPA-2 when subjected to freeze-drying was studied. Several additives were tested as protective agents against freeze-drying injury. Maximum viability of the bacterial cells was obtained with disaccharides (survival levels > 60%). Freeze-dried samples were rehydrated with several media; the highest percentage viability was obtained with 10% non-fat skim milk (100%+). The effect of initial bacterial load on the final recovery was dependent on protectant but not on rehydration media. Sucrose was an effective protectant when a high initial concentration (10(10) cfu ml(-1) was used; the opposite occurred with non-fat skim milk. The use of 10(10) cfu ml(-1) as an initial concentration, sucrose as a protectant and non-fat skim milk as a rehydration medium enabled 100% of P. agglomerans viability to be conserved after freeze-drying. Results suggest the possibility of achieving a good formulation system for the studied biocontrol agent with a high number of viable cells to be used toward pathogens, which is desirable for the industrial development of the product.

The effect of media, serum and temperature on in vitro survival of bovine blastocysts after Open Pulled Straw (OPS) vitrification

The recently introduced Open Pulled Straw (OPS) vitrification technique has successfully been used for cryopreserving porcine embryos as well as for bovine embryos and oocytes. The aim of this work is to investigate several factors on the in vitro survival of bovine blastocysts. In 5 experiments, a total of 862 in vitro produced blastocysts and expanded blastocysts was vitrified and wanned using the OPS technology, then cultured in vitro for an additional 3 days. The culture medium in Experiments 1 to 4 was SOFaa with supplements and 5% calf serum (CS). In Experiment 1, the replacement of TCM-199+20% CS with PBS+20% CS in the holding medium during vitrification and warming did not result in significant differences in the re-expansion (92 vs 95%) and hatching rates (79 vs 72%). In Experiment 2, the PBS holding medium was supplemented with either 20% CS, 5 mg/mL bovine serum albumin (BSA) or 3 mg/mL polyvinyl-alcohol (PVA). Although the re-expansion rates did not differ (98, 95 and 93%, respectively), there was a decrease in the hatching rate after vitrification with PVA (77 and 78 vs 51%, respectively). In Experiment 3, the influence of temperature of equilibration media prior to and rehydration media after the vitrification was investigated. When the temperature of these media was adjusted to 20 °C instead of the standard 35 °C, both the re-expansion and the hatching rates decreased markedly. However, increasing the time of equilibration with the diluted cryoprotectant solution at 20 °C eliminated these differences. In Experiment 4, the ethylene-glycol and dimethyl sulfoxide cryoprotectant mixture was replaced with ethylene glycol-ficoll-trehalose solution. No difference in the re-expansion (89 vs 96%, respectively) or hatching rate (79 vs 84%, respectively) was detected. In Experiment 5, the vitrified-warmed blastocysts were cultured in SOFaa medium supplemented with 5% CS or 5 mg/mL BSA. Although the re-expansion rates were identical in the 2 groups (95%), the hatching rates were lower when embryos were cultured in BSA (71 and 47%, respectively). These findings indicated the possible broader application for OPS, as they demonstrated that the physical advantages of rapid cooling and warming may be accompanied by different chemical composition (holding media, cryoprotective additives) according to the requirements of the biological structure. Our study also shows the need for serum supplementation of the medium for hatching to occur after OPS vitrification.

Влияние условий дегидратации и ионной силы среды регидратации на осмотический отклик и уровень лизиса регидратированных эритроцитов

Влияние условий дегидратации и ионной силы среды регидратации на осмотический отклик и уровень лизиса регидратированных эритроцитов

Effect of the rehydration medium on the recovery of freeze-dried lactic acid bacteria.

Sixteen cultures of lactic acid bacteria were freeze-dried in 10% nonfat skim milk plus 0.75 M adonitol and rehydrated by using different rehydration media. Marked variations in their capacity to repair cellular damage after freeze-drying were observed among the species and strains under consideration.

Effects of the Concentrations of Components of a Few Rehydration Media on the Viability of Freeze-Dried Marine Chemoheterotrophic Spirilla

Effects of the Concentrations of Components of a Few Rehydration Media on the Viability of Freeze-Dried Marine Chemoheterotrophic Spirilla

Effect of rehydration on recovery, repair, and growth of injured freeze-dried Salmonella anatum.

From 70 to 90% of the Salmonella anatum cells that survived freeze-drying in nonfat milk solids were injured. After rehydration, these injured survivors failed to grow on a selective plating medium containing deoxycholate but could form colonies on a nonselective medium. In a suitable environment after rehydration, injury disappeared in most of these cells. The rate of this repair at 25 C was very rapid initially and, in a medium containing milk solids, was completed within 1 hr after rehydration. The repaired cells initiated growth about 1 hr later than normal cells and grew at a slower rate. In a medium containing milk solids, initial recovery, extent of repair of injury, initiation of growth, and rate of growth were not influenced by supplementation with extra nutrients in other rehydration media. Rehydration controlled by modifying the concentrations of lactose, sucrose, or milk solids in the rehydration medium influenced the recovery of cells and the time that growth was initiated. Glycerol failed to increase recovery. Higher numbers of cells were recovered by rehydrating at 15 to 25 C, but an earlier initiation of growth and more rapid growth were observed at 35 C.

Effect of Rehydration on Recovery, Repair, and Growth of Injured Freeze-Dried Salmonella anatum1

From 70 to 90% of the Salmonella anatum cells that survived freeze-drying in nonfat milk solids were injured. After rehydration, these injured survivors failed to grow on a selective plating medium containing deoxycholate but could form colonies on a nonselective medium. In a suitable environment after rehydration, injury disappeared in most of these cells. The rate of this repair at 25 C was very rapid initially and, in a medium containing milk solids, was completed within 1 hr after rehydration. The repaired cells initiated growth about 1 hr later than normal cells and grew at a slower rate. In a medium containing milk solids, initial recovery, extent of repair of injury, initiation of growth, and rate of growth were not influenced by supplementation with extra nutrients in other rehydration media. Rehydration controlled by modifying the concentrations of lactose, sucrose, or milk solids in the rehydration medium influenced the recovery of cells and the time that growth was initiated. Glycerol failed to increase recovery. Higher numbers of cells were recovered by rehydrating at 15 to 25 C, but an earlier initiation of growth and more rapid growth were observed at 35 C.