Freeze drying of histocompatibility typing sera.

This report describes a unit employed for the freeze drying of histocompatibility typing serum using a 50-hr cycle. This unit will process approximately 3200 3-ml vials with a final residual moisture content of less than 2%. The system employs dry ice-alcohol cooled circulating baths to maintain the condensers below ?60 °C, and two shelf cooling baths to maintain the product at required temperatures during the freeze drying process.The results of a 5-yr study of the effect of residual moisture as a function of time and storage temperature is also included. Studies conducted to date indicate that with residual moistures below 2%, freeze dried histocompatibility sera can be stored at +4 °C without the loss of significant tissue typing factors. Solubility of all serum was lost when stored at +37 °C or higher during this same 5-year period.     

Influence of freeze-drying and spray-drying preservation methods on survivability rate of different types of protectants encapsulated Lactobacillus acidophilus FTDC 3081

ABSTRACT

The aims of this study were to compare the effectiveness of different drying methods and to investigate the effects of adding a series of individual protectant such as skim milk, sucrose, maltodextrin, and corn starch for preserving Lactobacillus acidophilus FTDC 3081 cells during spray and freeze-drying and storage at different temperatures. Results showed a remarkable high survival rate of 70–80% immediately after spray- and freeze-drying in which the cell viability retained at the range of 10⁹ to 10¹⁰ CFU/mL. After a month of storage, maltodextrin showed higher protective ability on both spray- and freeze-dried cells as compared to other protective agents at 4°C, 25°C, and 40°C. A complete loss in viability of spray-dried L. acidophilus FTDC 3081 was observed after a month at 40°C in the absence of protective agent.     

Survivability and storage stability of Trichoderma atroviride TRS40 preserved by fluidised bed drying on various agriculture by-products

Agriculture by-products were applied to proliferate biomass of Trichoderma atroviride TRS40 in solid state fermentation (SSF) cultures. The culture media overgrown with mycelium together with conidia were preserved by fluidised bed drying at various temperatures (50°C, 60°C and 70°C) and the received biopreparations were stored for 12 months. In order to determine the suitability of TRS40 in the production of biopreparations, the influence of preservation process and storage time on their survivability was examined. The three-component mixture proved more effective in the SSF cultures, ensuring TRS40 count at 6.07?×?10^9?CFU/g?dm, which was ca. 6 times higher than in the mono-component medium. TRS40 survivability after preservation at various temperatures ranged from 40.4% to 100%, regardless of carrier type. In turn, after 12-month storage of the biopreparations produced on the three-component medium, regardless of drying temperature, the number of viable cells ranged from 2.43?×?10⁸ to 2.49?×?10^8?CFU/g?dm. Furthermore, selected parameters of growth kinetics in the Bioscreen C system were determined. The storage time of biopreparations had various effects on growth kinetic parameters. In addition, the preserved preparations based on the TRS40 retained their capability for biosynthesis of hydrolases, even after 12 months of storage.     

Changes in quality of Phellinus gilvus mushroom by different drying methods

This study was conducted to investigate the changes in characteristics of the Phellinus gilvus mushroom as influenced by drying methods after harvest. The lowest weight loss rate of P. gilvus mushroom was 75.8% with drying in the shade and 80% by dryer (60°C). The size loss rate of pileus was 19.3% of that in a hot air dryer (60°C). The hardness of dried material context using a hot air dryer (60°C) was the lowest (20 kg/cm²), and that by a dry oven (60°C) was the highest (457 kg/m²). For ΔE value, 4.9 of context and 2.6 of tubes using drying in the shade (20°C) were found to be the lowest. The survival rate of sarcoma 180 treated with P. gilvus dried in the sun was the lowest (51.8%), and this was considered the most effective method for antitumor activity against sarcoma 180.     

Box–Behnken analysis and storage of spray-dried collagenolytic proteases from Myceliophthora thermophila submerged bioprocess

Enzymes do not have long-term storage stability in soluble forms, thus drying methods could minimize the loss of enzymatic activity, the spray dryer removes water under high temperatures and little time. The aims of this study were to improve the stability of enzymatic extract from Myceliophthora thermophila for potential applications in industry and to evaluate the best conditions to remove the water by spray drying technique. The parameters were tested according to Box–Behnken and evaluated by analysis of variance (ANOVA), all the parameters measured were found to influence the final enzyme activity and spray drying process yield ranged from 38.65 to 63.75%. Enzyme powders showed increased storage stability than extract and maintained about 100% of collagenolytic activity after 180 days of storage at 30°C. The results showed that the microbial enzymes maintained activity during the spray drying process and were stable during long-term storage; these are promising characteristics for industrial applications.     

Effect of drying temperature on carrageenan yield and quality of Kappaphycus alvarezii (Rhodophyta,Solieriaceae) cultivated in Brazil

This study evaluates the effect of different drying temperatures on the properties of semi-refined (SR) and refined (R) carrageenan extracted from Kappaphycus alvarezii cultivated in Brazil. Drying kinetics was studied in seaweeds under the following treatments: sun drying (control) and drying at 40, 60, and 90 °C in convective air dryer. Drying was carried out until the moisture content of seaweeds reached values below of 30 % on wet basis. Significant reductions in drying time were observed with the increase of temperature. At 90 °C, 30 % moisture content was reached in 100 min, as compared with the 1,440 min required by the sun-drying treatment. SR yields showed no significant differences when compared to the control, varying from 40 to 44 %, while R had a significantly higher yield (30 %) at 90 °C in relation to control (26 %). Gel strength of SR was significantly higher in the sun-dried samples (1,685.1 g cm^?2) and 60 °C samples (1,727.2 g cm^?2), but no significant differences were observed in R gel strengths. Lowest syneresis was observed in both SR (9.8 %) and R (10.3 %) after the treatment at 90 °C. Significantly lower viscosity values were observed for SR at 60 °C (233 mPa s) and at 90 °C (175 mPa s), as for R, the lowest value was observed at 90 °C (205 mPa s). Based on these results, it was concluded that best results for both types of carrageenan are obtained drying at 60 °C.     

Long-term stability of oxidative stress biomarkers in human serum

The storage time and storage temperature might affect stability of oxidative stress biomarkers, therefore, they have to be analyzed after long-term storage of serum samples. The stability of three biomarkers reflecting oxidative stress: reactive oxygen metabolites (ROM) for hydroperoxides, total thiol levels (TTL) for the redox status and biological antioxidant potency (BAP) for the antioxidant status, was investigated at several time points during 60 months of storage at ?20 and ?80?°C. Biomarkers ROM and BAP showed a very good stability during storage for 60 months at both temperatures. In addition, the correlation of the data after 60 months of storage compared with the starting data was very good with correlation coefficients >0.9. The TTL assay showed good results in serum samples stored at ?80?°C, but not in samples stored at ?20?°C. Serum samples for analysis of the set of oxidative stress biomarkers ROM, BAP and TTL can be stored up to 60 months at ?80?°C. ROM and BAP can also be stored at ?20?°C during this period. The present results are very important for the biomarker-related epidemiological studies that make use of biobanks with samples stored for many years and for new project planning, including sample storage conditions.     

Evaluation of functional stability of quercetin as a raw material and in different topical formulations by its antilipoperoxidative activity

The present study evaluates the antioxidant activity of the flavonol quercetin, and its functional stability as a raw material and when added in formulations. The iron-chelating activity was determined using the bathophenanthroline assay, and the functional stability was evaluated with the antilipoperoxidative assay. Raw material presented concentration-dependent antilipoperoxidative and iron-chelating activities. The initial antilipoperoxidative activity of the raw material, cream and gel-cream were 63%, 78%, and 69%, respectively. There was no detectable loss of activity during 182 days (6 months) of storage at all tested temperatures (4°C, room temperature [RT], 37°C, and 45°C) for the raw material. Considering the method variability of 10%, activity loss greater than 10% for nonionic cream was detected after 126 days at 4°C (20.1%), decreasing thereafter to 22.2% after 182 days. At 45°C, the loss of activity started after 182 days (13.2%). For the anionic gel-cream, activity loss started after 84 days (28.4%, 45°C), decreasing after 182 days to 40.3% at 45°C. At 37°C, activity loss was detected after 182 days (12%). In conclusion, the results suggest that the activity of quercetin depends on iron chelation, and its posible usefulness as a topical antioxidant to prevent oxidative stress-induced skin damage depends on maintaining its antilipoperoxidative activity stored at RT, which avoids special storage conditions. Published: February 3, 2006     

Yeasts preservation: alternatives for lyophilisation

The aim of the study was to compare the effect of two low-cost, low technology traditional methods for drying starter cultures with standard lyophilisation. Lyophilised yeast cultures and yeast cultures preserved in dry rice cakes and dry plant fibre strands were examined for viable cell counts during 6?months storage at 4 and 25?°C. None of the yeast cultures showed a significant loss in viable cell count during 6?months of storage at 4?°C upon lyophilisation and preservation in dry rice cakes. During storage at 25?°C in the dark, yeast cultures preserved in dry rice cakes, and lyophilised cultures of Saccharomyces cerevisiae and Issatchenkia orientalis showed no significant loss of viable cells up to 4?months of storage. Yeast cultures preserved in dry plant fibre strands had the greatest loss of viable count during the 6?months of storage at 25?°C. Preservation of yeasts cultures in dry rice cakes provided better survival during storage at 4?°C than lyophilisation. The current study demonstrated that traditional methods can be useful and effective for starter culture preservation in small-scale, low-tech applications.     

Viability of rape (Brassica napus L.) seeds following selection on the basis of newly-emerged radicles then subsequent drying and storage

Germinating rape seeds selected on the basis of newly-emerged radicles (1 ± 0.5 mm) were dried to an equilibrium moisture content (c. 11%) in air at 20°C and 80% relative humidity without loss of viability. Storage life of these low-moisture-content germinating (LMCG) seeds at 15°C was limited to 7 days before viability was significantly reduced. However, viability of LMCG seeds was maintained for 84 days in storage at -20°C. Longer periods in store reduced viability, but 96% of seeds still remained viable after 336 days at - 20°C. Increasing periods of storage at -20°C reduced the subsequent seed longevity at 15°C, indicating a reduction in vigour during storage. Storage under reduced pressure or in a nitrogen atmosphere had little significant effect on seed longevity. Reduction of moisture content below 11% using vacuum drying at a range of temperatures reduced seed vigour.     

Cryopreservation and microencapsulation of a probiotic in alginate-chitosan capsules improves survival in simulated gastrointestinal conditions

The aim of the present study was to focus on the impact of two different methods and the effects of cryoprotectants on the survival of a probiotic bacterium, Streptococcus phocae PI80, during storage. For the protection of freeze dried cells, the optimal storage conditions were determined with a high survival rate. After the freeze drying process, all cryoprotectants exhibited a protective effect on cell viability at all storage temperatures. High relative cell viability was observed when cells were incubated at ?20°C, which was optimum for the protection of S. phocae PI80. Trehalose was the most promising cryoprotectant at all temperatures during the storage period of bacterial cells. The combination of trehalose + skim milk showed more than 85% survivability compared to other combinations at ?20°C for 60 days. In addition, encapsulation of probiotic cells into alginate-chitosan gel capsules showed better survival of S. phocae cells (5.468 ± 0.15 LogCFU/mL) with high bacteriocin activity at ?20°C for six months. The cell-loaded microcapsules remained stable when treated with simulated gastric and intestinal fluids. After 6 h in vivo treatment, the capsules were found to be broken, releasing the probiotic cells directly into the intestinal system of rats. Therefore, microencapsulation was found to be the most efficient technique, which not only protected the cells for a longer time but also released the cells into the in vivo intestinal system.     

Flow cytometric analysis from fish samples stored at low,ultra-low and cryogenic temperatures

Flow cytometry is a valuable tool in biomedical and animal sciences. However, equipment used for such analysis presents limitations at field conditions, suggesting then preservation procedures for future analysis at laboratory conditions. In this study, freezing at low (−20 °C), ultra-low (−80 °C) and cryogenic temperatures (−196 °C, i.e. liquid nitrogen) were used as preservation procedures of fish tissue. Samples were maintained in 0.9% NaCl or lysing solution, and stored at the temperatures above for 0 (fresh control), 60, 120 and 180 days of storage. After storage, the samples were thawed and proceeded to flow cytometric analysis. Storage at low temperatures (−20 °C), both in lysing and 0.9% NaCl, exhibited poor results when analyzed after 60, 120 and 180 days, showing noisy peaks, deviation in the DNA content and absence of peaks. Ultralow (−80 °C) and cryogenic (−196 °C) temperatures, both in lysing solution and 0.9% NaCl, showed good results and high quality of histograms. Both storage procedures gave similar histograms and DNA content in comparison with control group (fresh) even after 60, 120 and 180 days of storage, exhibiting the main peak at 2C content from diploid cells and a secondary peak at 4C derived from dividing cells. In conclusion, samples may be stored for 180 days at −80 °C and −196 °C in both, 0.9% NaCl or lysing solution. As cryogenic temperatures in liquid nitrogen permits indefinite storage, this procedure should be used for long-term preservation.     

Effect of water content and temperature on seed longevity of seven Brassicaceae species after 5 years of storage

Maximising seed longevity is crucial for genetic resource preservation and longevity of orthodox seeds is determined by environmental conditions (water content and temperature). The effect of water content (down to 0.01 g·H₂O·g^?1) on seed viability was studied at different temperatures for a 5‐year storage period in taxonomically related species. Seeds of seven Brassicaceae species (Brassica repanda, Eruca vesicaria, Malcolmia littorea, Moricandia arvensis, Rorippa nasturtium‐aquaticum, Sinapis alba, Sisymbrium runcinatum) were stored at 48 environments comprising a combination of eight water contents, from 0.21 to 0.01 g·H₂O·g^?1 DW and six temperatures (45, 35, 20, 5, ?25, ?170 °C). Survival curves were modelled and P50 calculated for those conditions where germination was reduced over the 5‐year assay period. Critical water content for storage of seeds of six species at 45 °C ranged from 0.02 to 0.03 g·H₂O·g^?1. The effect of extreme desiccation at 45 °C showed variability among species: three species showed damaging effects of drying below the critical water content, while for three species it was neither detrimental nor beneficial to seed longevity. Lipid content could be related to longevity, depending on the storage conditions. A variable seed longevity response to water content among taxonomically related species was found. The relative position of some of the species as long‐ or short‐lived at 45 °C varied depending on the humidity at which storage behaviour was evaluated. Therefore, predictions of survival under desiccated conditions based on results obtained at high humidity might be problematic for some species.     

The response of bacterial spores to vacuum treatments. II. Germination and viability studies.

The viability of Bacillus megaterium spores has been determined after exposure to vacuum dehydration at temperatures between 0 and 65 °C, for periods up to 24 hr. A curvilinear relationship has been demonstrated between viability and drying temperature, with minimum viability occuring around 15 °C and increases in viability being shown above 35 °C. In contrast to vegetative bacteria, reequilibration of the dried spores to 2 × 10^?3 or 10 Torr aqueous vapor pressure, and/or subsequent exposure to oxygen had no effect on viability. Dehydration, rehydration and oxygen treatments had no effect on the time for outgrowth of the spores or on the growth rate of the resultant vegetative cells. Physical loss of spores from samples was not demonstrated during any of these treatments. Evidence has been presented for a novel type of spore activation, which occurs during vacuum dehydration at high temperatures, to an extent that is dependent upon drying time. The mechanism of this activation is unlike that of conventional heat or chemical activation but is oxygen independent and unaffected by reequilibration to 2 × 10^?3 or 10 Torr.     

Investigations into the effects of processing on the retention of the carotenoid fractions of Leucaena leucocephala during storage,and the effects of processing on mimosine concentration

Leucaena leucocephala leaves were subjected to several drying treatments followed by pelleting and/or inclusion of ethoxyquin as antioxidant and analysed monthly to determine rates of loss of carotenes and xanthophylls. Rapid sun drying yielded leucaena leaf meal with carotene and xanthophyll concentrations of 446 and 865 mg/kg, respectively, but considerable losses occurred during drying at 60 and 145°C, and as a result of blanching in hot water or ferrous sulphate solution. Loss rates of carotenes during storage were in the range 19–40 mg/kg per month, and of xanthophylls, 29–53 mg/kg per month. Carotenoids in sun-dried leaf meal were most stable during storage. Pelleting or inclusion of ethoxyquin did not reduce carotenoid loss rates below those of unpelleted leaf meal.The mimosine content of untreated sun-dried leucaena leaf meal was 3.2%, but the concentration decreased to 2.5 and 1.8% on drying at 60 and 145°C, respectively. The concentrations of mimosine in sun-dried leafmeals of water-blanched and ferrous sulphate-blanched leucaena were 2.0 and 2.1%, respectively. Treatments causing maximum losses of mimosine were those which brought about maximum losses of carotenoids.     

The protection,stabilization and reactivation of estradiol receptors in human myometrial cytosol

Protection of estradiol receptor binding sites in human mymetrial cytosol is achieved though the agency of dithiothreitol (Cleland's reagent) (lmM) at 4°C and ? 20°C storage temperatures. Prolonged storage of cytosol at 4° C without the protective reagent results in substantial loss of binding activity. This is partially restored after Cleland's reagent (lmM) addition. However, cytosol inactivated by heating at 60°C for 30 min cannot be reconstituted in this way.     

Heat treatments for the control of sprouting of potatoes during storage

Potatoes were given hot water dip and vapour heat treatments at temperatures ranging from 50 to 80°C and 60 to 70°C respectively for various durations to control sprouting. Hot water treatment at its effective temperatures and durations resulted in undesirable physical damage to tubers. On the other hand, vapour heat treatment at 60°C with 95 ° 5 r.h. for 60 min suppressed sprouting, when given to tubers at the sprout emergence stage, without causing any deleterious side effects. The treatment, when repeated after 3 wk further reduced sprout yield during storage at ambient conditions (22–35°C, 50–80 r.h.). Comparative studies on the efficacy of vapour heat treatment and manual de-sprouting showed that vapour heat was superior to manual de-sprouting.     

Effect of freezing and freeze-drying on the viability and storage of Lilium longiflorum L. and Zea mays L. pollen

The freeze-preservation of pollen is dependent on the interaction of several factors such as freezing rate, thawing rate, freeze-drying temperature and duration, storage temperature and environment and rehydration rates. Changes in any of these variables affects the others directly or indirectly.Rapid freezing of pollen at rates of approximately 200 °C/min maintains the highest degree of viable pollen in combination with rapid thawing rates of 218 °C/min. Rapid cooling and slow rewarming resulted in a substantial loss of pollen viability. This might indicate that intracellular ice crystals formed during rapid cooling perhaps grow into larger ice masses during slow rewarming or storage at temperatures above ?50 °C.The germinability of pollen freeze-dried at temperatures below ?50 °C was also prolonged over that of the controls. Germination values for unfrozen pollen stored for 30 days at 0–5 °C averaged 50% for lily and 20% for corn. Freeze-dried pollen stored for 30 days at the same temperature yielded considerably higher viability percentages for both lily and corn pollen. Drying time is an important factor, perhaps indicating that residual moisture is critical. Freeze-dried pollen can be stored at higher temperatures than frozen and control pollen. Freeze-dried material stored for five months at 0–5 °C, upon slow rehydration yielded intact grains which has average germination percentages of 25 for lily and 15 for corn. The same pollen upon rapid rehydration showed rupturing of 20–40% of the cells and practically no germination.     

Microcosm evaluation of bioaugmentation after field-scale thermal treatment of a TCE-contaminated aquifer

This paper investigates effects of combining thermal and biological remediation, based on laboratory studies of trichloroethene (TCE) degradation. Aquifer material was collected 6 months after terminating a full-scale Electrical Resistance Heating (ERH), when the site had cooled from approximately 100°C to 40°C. The aquifer material was used to construct bioaugmented microcosms amended with the mixed anaerobic dechlorinating culture, KB-1^TM, and an electron donor (5 mM lactate). Microcosms were bioaugmented during cooling at 40, 30, 20, and 10°C, as temperatures continually decreased during laboratory incubation. Redox conditions were generally methanogenic, and electron donors were present to support dechlorination. For microcosms bioaugmented at 10°C and 20°C, dechlorination stalled at cis-dichloroethene (cDCE) and vinyl chloride (VC) 150 days after bioaugmentation. However, within 300 days of incubation ethene was produced in the majority of these microcosms. In contrast, dechlorination was rapid and complete in microcosms bioaugmented at 30°C. Microcosms bioaugmented at 40°C also showed rapid dechlorination, but stalled at cDCE with partial VC and ethene production, even after 150 days of incubation when the temperature had decreased to 10°C. These results suggest that sequential bioremediation of TCE is possible in field-scale thermal treatments after donor addition and bioaugmentation and that the optimal bioaugmentation temperature is approximately 30°C. When biological and thermal remediations are to be applied at the same location, three bioremediation approaches could be considered: (a) treating TCE in perimeter areas outside the source zone at temperatures of approximately 30°C; (b) polishing TCE concentrations in the original source zone during cooling from approximately 30°C to ambient groundwater temperatures; and (c) using bioremediation in downgradient areas taking advantages of the higher temperature and potential release of organic matter.     

Unraveling protein stabilization mechanisms: Vitrification and water replacement in a glass transition temperature controlled system

The aim of this study was to elucidate the role of the two main mechanisms used to explain the stabilization of proteins by sugar glasses during drying and subsequent storage: the vitrification and the water replacement theory. Although in literature protein stability is often attributed to either vitrification or water replacement, both mechanisms could play a role and they should be considered simultaneously. A model protein, alkaline phosphatase, was incorporated in either inulin or trehalose by spray drying. To study the storage stability at different glass transition temperatures, a buffer which acts as a plasticizer, ammediol, was incorporated in the sugar glasses. At low glass transition temperatures (< 50 °C), the enzymatic activity of the protein strongly decreased during storage at 60 °C. Protein stability increased when the glass transition temperature was raised considerably above the storage temperature. This increased stability could be attributed to vitrification. A further increase of the glass transition temperature did not further improve stability. In conclusion, vitrification plays a dominant role in stabilization at glass transition temperatures up to 10 to 20 °C above storage temperature, depending on whether trehalose or inulin is used. On the other hand, the water replacement mechanism predominately determines stability at higher glass transition temperatures.