Preservation of coagulation efficiency ofMoringa oleifera, a natural coagulant

In recent years, there has been an interest to useMoringa oleifera as the natural coagulant due to cost, associated health and environmental concerns of synthetic organic polymers and inorganic chemicals. However, it is known thatM. oleifera as the natural coagulant is highly biodegradable and has a very short shelf life. This research was carried out to investigate the effects of storage temperature, packaging methods, and freeze-drying on the preservation ofM. oleifera seeds powders. Non freeze-driedM. oleifera was prepared into different packaging namely open container, closed container and vacuum packing, whilst, freeze-driedM. oleifera was stored in closed container and vacuum packing. Each of the packaging was stored at room temperature (30 to 32°C) and refrigerator (4°C). The turbidity removal efficiencies of storedM. oleifera were examined using jar test at monthly interval for 12 months. The results indicated that non freeze-driedM. oleifera kept in the refrigerator (4°C) would preserve its coagulation efficiency. In addition, closed container and vacuum packing were found to be more appropriate for the preservation of non freeze-driedM. oleifera, compared to open container. Freeze-driedM. oleifera retained its high coagulation efficiency regardless the storage temperature and packaging method for up to 11 months. Besides, higher increment in zeta potential values for water coagulated with freeze-driedM. oleifera indicated the higher frequency of charge neutralization and better coagulation efficiency of freeze-driedM. oleifera, compared to non freeze-dried seeds. As a coagulant,M. oleifera did not affect the pH of the water after treatment.     

Inbound container storage price competition between the container terminal and a remote container yard

With the rapid development of global ocean transportation, storage space in container terminals is becoming a scarce resource. Hence, the terminal yard only performs as a temporary storage facility for inbound cargos. A storage charge is levied for inbound cargos that stay longer than a free storage time (called free-time-limit). After the free-time-limit, customers may move cargos from the terminal yard to a remote container yard where the storage price is lower than that in the terminal. This paper proposes inbound container storage pricing game models between the container terminal and a remote container yard. Two cases are considered: (1) the inbound container’s dwell time is random and follows a probability distribution function; (2) the inbound container’s dwell time is sensitive to the storage prices. The primary objective of this paper is to analyze the storage pricing behavior and competition outcomes of the container terminal and the remote container yard. A number of insights and analysis are provided.     

Evaluation and utility of new CE marked containers (CELLFLEX- MacoPharma) for bone bank

In order to guarantee the required level of quality for our Bone & Tissue Banking, we evaluated a new CE marked container (CELLFLEX MacoPharma), for packing, transport, processing and storage of bones fortherapeutic use. The use of CE marked containers is mandatory for organ and tissue containers (Medical Device Directive 93/42). We carried out a three-phase study: (1)Evaluation, (2)Implementation, (3)Audit The product was evaluated for the following criteria: Dash mechanical resistance, Dash air tightness, Dash fragility, Dash capacity. No damage was observed after the storage period, even after immersion in liquid nitrogen. No breakages were observed after provoked impact tests (pots dropped onto the floor). The pot capacity evaluation showed that the inner pot volume (∼500 ml) permits adequate storage of tendons and the majority of bone allografts. In conclusion, this evaluation has shown that the CELLFLEX kit is suitable for long duration preservation of bone allografts even at very low temperature conditions (vapour phase nitrogen). Its format and structure permit preservation of most bone allografts. This revised version was published online in July 2006 with corrections to the Cover Date.     

Pricing storage of outbound containers in container terminals

In container yard of container terminals, a storage charge is imposed to encourage customers to limit the space required for their containers. This study addresses the storage price scheduling problem for the storage of outbound containers. The price schedule consists of the free-time limit, which is the maximum duration for a container to stay in the yard without any charge, and storage charge per day for storing a container past the free-time-limit. Empirical data suggests that the efficiency of loading operations significantly depends on the space utilized by a vessel’s outbound containers. Mathematical models are developed to determine the optimal storage price schedule in such a manner that the terminal’s total profit is maximized or the total system’s cost is minimized. Both single and multi-vessel cases are considered. Properties of the optimal solution are derived from the mathematical models and numerical experiments are conducted to validate solutions.     

Effects of storage conditions on the stability and distribution of clinical trace elements in whole blood and plasma: Application of ICP-MS

BackgroundKnowledge of trace element stability during sample handling and preservation is a prerequisite to produce reliable test results in clinical trace element analysis.MethodAn alkaline dissolution method has been developed using inductively coupled plasma mass spectrometry to quantify eighteen trace element concentrations: vanadium, chromium, manganese, cobalt, nickel, copper, zinc, arsenic, selenium, bromine, molybdenum, cadmium, antimony, iodine, mercury, thallium, lead, and bismuth in human blood, using a small sample volume of 0.1 mL. The study evaluated the comparative effects of storage conditions on the stability of nutritionally essential and non-essential elements in human blood and plasma samples stored at three different temperatures (4 °C, −20 °C and −80 °C) over a one-year period, and analysed at multiple time points. The distribution of these elements between whole blood and plasma and their distribution relationships are illustrated using blood samples from 66 adult donors in Queensland.ResultsThe refrigeration and freezing of blood and plasma specimens proved to be suitable storage conditions for many of the trace elements for periods up to six months, with essentially unchanged concentrations. Substantially consistent recoveries were obtained by preserving specimens at −20 °C for up to one year. Ultra-freezing of the specimens at −80 °C did not improve stability; but appeared to result in adsorption and/or precipitation of some elements, accompanied by a longer sample thawing time. A population sample study revealed significant differences between the blood and plasma concentrations of six essential elements and their relationships also varied significantly for different elements.ConclusionBlood and plasma specimens can be reliably stored at 4 °C for six months or kept frozen at −20 °C up to one year to obtain high quality test results of trace elements.     

Preservation of basidiomycete strains on perlite using different protocols

For preservation of 31 basidiomycete strains on perlite in cryovials we used five different perlite protocols to compare their applicability in laboratories with different equipment, namely a viability of the controlled freezing device or the electric deep-freezer and liquid nitrogen supply. The viability of the strains, macromorphological characteristics and the production of laccase were tested after 48 h, six months and one year of storage in the respective device. Our results indicated that the different response to the freezing/thawing process is an intrinsic feature of the respective strain. Nevertheless, the highest viability and preservation of laccase production in our tested strains was found when we used pre-freezing to −80 °C at a freezing rate of 1 °C/min in a programmable IceCube 1800 freezer or in freezing container Mr. Frosty before storage in liquid nitrogen or at ultra-low temperature freezer at −80 °C, respectively. The two abovementioned protocols enable all tested strains to survive three successive freezing/thawing cycles without substantial reduction of growth rate. The majority of the strains also do not lose laccase production. Our results showed that direct immersion of the strains into liquid nitrogen or placing them into −80 °C without pre-freezing is not suitable for basidiomycete cryopreservation.     

火龙果果实的功效及保鲜贮存技术研究(综述)

火龙果果实含有丰富的糖、白蛋白、不饱和脂肪酸、膳食纤维、维生素及多种微量元素,在抗氧化、降血脂、美容养颜等方面具有较好的保健功效,开发利用前景广阔。本文综述火龙果果实的营养成分和保健价值以及在保鲜贮存方面的研究进展,以期为火龙果的生产和科研提供参考依据。     

The relationship between fungal preservation method and secondary metabolite production in Metarhizium anisopliae and Fusarium oxysporum

The effects of preservation regime on secondary metabolite production in two genera of economically important fungi, Metarhizium anisopliae and Fusarium oxysporum, was assessed using thin layer chromatography and high performance liquid chromatography over a 2-year testing period. Five preservation regimes, commonly used in microbial culture collections throughout the world were investigated: continual sub-culture, lyophilization, storage of mycelial plugs in water, storage at –20 °C and cryopreservation with liquid nitrogen. Preservation regime influenced secondary metabolite production in the test fungi. Changes in secondary metabolite profiles occurred after relatively short storage periods in most strains, irrespective of the preservation treatment used. Although no preservation treatment can be guaranteed to provide total stability of secondary metabolite production, cryopreservation was the best of the methods tested. Response to preservation and storage also differed between strains of the same species. Therefore, there is a need to develop new and existing preservation criteria with an emphasis on strain-specific criteria in order to reduce the prospects of instability in secondary metabolite production.     

Preservation of H2 production activity in nanoporous latex coatings of Rhodopseudomonas palustris CGA009 during dry storage at ambient temperatures

To assess the applicability of latex cell coatings as an ‘off‐the‐shelf’ biocatalyst, the effect of osmoprotectants, temperature, humidity and O₂ on preservation of H₂ production in Rhodopseudomonas palustris coatings was evaluated. Immediately following latex coating coalescence (24 h) and for up to 2 weeks of dry storage, rehydrated coatings containing different osmoprotectants displayed similar rates of H₂ production. Beyond 2 weeks of storage, sorbitol‐treated coatings lost all H₂ production activity, whereas considerable H₂ production was still detected in sucrose‐ and trehalose‐stabilized coatings. The relative humidity level at which the coatings were stored had a significant impact on the recovery and subsequent rates of H₂ production. After 4 weeks storage under air at 60% humidity, coatings produced only trace amounts of H₂ (0–0.1% headspace accumulation), whereas those stored at < 5% humidity retained 27–53% of their H₂ production activity after 8 weeks of storage. When stored in argon at < 5% humidity and room temperature, R. palustris coatings retained full H₂ production activity for 3 months, implicating oxidative damage as a key factor limiting coating storage. Overall, the results demonstrate that biocatalytic latex coatings are an attractive cell immobilization platform for preservation of bioactivity in the dry state.     

Differences in preservation of canine chilled semen using different transport containers

In the present study, the effect of three different containers in the preservation of dog chilled semen, during 24, 48 and 72h was evaluated. Weekly sperm pools of different dogs were obtained, during 10 consecutive weeks. Semen samples were diluted in egg-yolk-Tris-fructose extender and stored in a Styrofoam box, a common Thermos flask and an Equitainer. Progressive motility, morphology and sperm membrane integrity were examined in semen aliquots taken daily from each container during the 3 days of storage. Additionally, integrity of the acrosome and sperm plasma membranes, determined by PI/Fitc-PSA staining was assessed at 48 and 72h of storage. At 24h no differences were observed between the three containers for the evaluated parameters. At 48h samples kept in the Equitainer presented a higher progressive motility than samples kept in the Thermos. At 72h, progressive motility was higher in the Equitainer than in the other two containers. Only samples kept in the Equitainer maintained similar levels of progressive motility between 24 and 72h. Membrane integrity assessed by eosin-nigrosin deteriorated over the 72h period, whereas functional membrane integrity determined by the hypoosmotic swelling test was independently affected by type of container (the Equitainer) kept a higher percentage of sperm cells with intact membrane) and time of storage (a decrease of membrane integrity between 24 to 72h). Staining with PI-Fitc-PSA allowed the detection of differences between containers but not between the two studied storage periods (48 and 72h). The results indicated that the use of the Equitainer is preferable when transporting chilled dog semen for more than 48h.     

A Novel Device for the Collection, Storage, Transport, and Delivery of Beneficial Insects, and its Application to Ophraella communa (Coleoptera: Chrysomelidae)

A multi-use device was developed for the collection, short-term storage, transport, and delivery of Ophraella communa (Coleoptera: Chrysomelidae), a biological control agent of common ragweed, Ambrosia artemisiifolia. The device is made from a 125-ml plastic specimen container that can hold O. communa adults or pupae. When used as an aspirator, insect collection and counting times are reduced. O. communa adults and pupae can be stored inside the container at 3°C with median survival of 41 and 21 days, respectively. A cotton wick saturated with water or a 5% sugar solution nourishes insects during transport and the container design minimizes insect mortality by providing an optimum microclimate during insect storage and transport. Designed to protect insects from rainfall and to limit encounters with predators and parasites, the containers can be used for field releases of O. communa adults and pupae. Although the container has been designed and tested for O. communa, it is highly versatile and could possibly be used with a variety of insect species.     

The Preservation of Bacteria and Fungi on anhydrous Silica Gel: an Assessment of Survival over Four years

The preservation method of Perkins (1962) using suspensions in skim-milk was used to preserve 33 bacteria and 22 fungi on anhydrous silica gel. During storage at room temperature, 64% of the bacteria and 77% of the fungi survived 1 year or more. Storage at 4° often increased the survival period c. 2- to 3-fold: 73% of the bacteria and all 12 of the fungi tested at 4° survived > 1 year. At the last testing, 60% of the bacteria and 36% of the fungi were still viable after storage at 4° for periods between 3 and 4 years. The Gram positive bacilli tended to survive the silica gel preservation process better than most Gram negative bacilli. Some factors influencing survival after preservation on silica gel are discussed; the results support the use of a closed storage tube.     

Comparison of fecal preservation and extraction methods for steroid hormone metabolite analysis in wild crested macaques

Since the non-invasive field endocrinology techniques were developed, several fecal preservation and extraction methods have been established for a variety of species. However, direct adaptation of methods from previous studies for use in crested macaques should be taken with caution. We conducted an experiment to assess the accuracy and stability of fecal estrogen metabolite (E1C) and glucocorticoid metabolite (GCM) concentrations in response to several preservation parameters: (1) time lag between sample collection and fecal preservation; (2) long-term storage of fecal samples in 80% methanol (MeOH) at ambient temperature; (3) different degrees of feces drying temperature using a conventional oven; and (4) different fecal preservation techniques (i.e., freeze-drying, oven-drying, and field-friendly extraction method) and extraction solvents (methanol, ethanol, and commercial alcohol). The study used fecal samples collected from crested macaques (Macaca nigra) living in the Tangkoko Reserve, North Sulawesi, Indonesia. Samples were assayed using validated E1C and GCM enzyme immunoassays. Concentrations of E1C and GCM in unprocessed feces stored at ambient temperature remained stable for up to 8 h of storage after which concentrations of both E1C and GCM changed significantly compared to controls extracted at time 0. Long-term storage in 80% MeOH at ambient temperature affected hormone concentrations significantly with concentrations of both E1C and GCM increasing after 6 and 4 months of storage, respectively. Drying fecal samples using a conventional oven at 50, 70, and 90 °C did not affect the E1C concentrations, but led to a significant decline for GCM concentrations in samples dried at 90 °C. Different fecal preservation techniques and extraction solvents provided similar results for both E1C and GCM concentrations. Our results confirm previous studies that prior to application of fecal hormone analysis in a new species, several preservation parameters should be evaluated for their effects on hormone metabolite stability. The results also provide several options for fecal preservation, extraction, and storage methods that can be selected depending on the condition of the field site and laboratory.     

Effects of storage conditions of Moringa oleifera seeds on its performance in coagulation

Moringa oleifera is a plant whose seeds have coagulation properties for treating water and wastewater. In this study the coagulation efficiency of Moringa oleifera kept in different storage conditions were studied. The Moringa oleifera seeds were stored at different conditions and durations; open container and closed container at room temperature (28 degrees C) and refrigerator (3 degrees C) for durations of 1, 3 and 5 months. Comparison between turbidity removal efficiency of Moringa oleifera kept in refrigerator and room temperature revealed that there was no significant difference between them. The Moringa oleifera kept in refrigerator and room temperature for one month showed higher turbidity removal efficiency, compared to those kept for 3 and 5 months, at both containers. The coagulation efficiency of Moringa oleifera was found to be dependent on initial turbidity of water samples. Highest turbidity removals were obtained for water with very high initial turbidity. In summary coagulation efficiency of Moringa oleifera was found independent of storage temperature and container, however coagulation efficiency of Moringa oleifera decreased as storage duration increased. In addition, Moringa oleifera can be used as a potential coagulant especially for very high turbidity water.     

Effects of preservation on pigmentation and length measurements in larval lampreys

The effects of two methods of preservation (fixation and storage in 10% formalin, and fixation in 10% formalin followed by storage in 95% alcohol) on pigmentation and morphometric features used for identification of larval Ichthyomyzon lampreys were analysed. Both short‐term (3 weeks) and long‐term (6 months) studies were conducted using digital analysis of images of fresh and preserved lampreys. Six standard morphometric lengths and 10 areas of pigmentation were analysed. All measurements were significantly affected by preservation. Preservative type affected pigmentation and morphometric characteristics differently, and characters were affected to different degrees. Multiple measurements over time showed that almost all changes occurred within 3 weeks of preservation. Regression equations allowed for accurate correction of preservation effects on morphometric measurements, but the effects on pigmentation levels were less predictable. Effects of preservation on larval lampreys need to be considered when comparing fresh and preserved specimens because they influence critical identification features.     

Natural biopolymer for preservation of microorganisms during sampling and storage

Stability of microbial cultures during sampling and storage is a vital issue in various fields of medicine, biotechnology, food science, and forensics. We have developed a unique bacterial preservation process involving a non-toxic, water-soluble acacia gum polymer that eliminates the need for refrigerated storage of samples.The main goal of this study is to characterize the efficacy of acacia gum polymer for preservation of pathogenic bacteria (Bacillus anthracis and methicillin-resistant Staphylococcus aureus—MRSA) on different materials, used for swabbing and filtration: cotton, wool, polyester, rayon, charcoal cloth, and Whatman paper.Acacia gum polymer used for preservation of two pathogens has been shown to significantly protect bacteria during dehydration and storage in all tested samples at the range of temperatures (5-45 °C for MRSA and 40-90 °C for B. anthracis). Our results showed higher recovery as well as higher viability during the storage of both bacteria in all materials with acacia gum. Addition of acacia gum polymer to swabbing materials or filters will increase efficacy of sample collection and identification of pathogenic bacteria from locations such as hospitals or the environment. Proposed approach can also be used for long-term storage of culture collections, since acacia gum contributes to viability and stability of bacterial cultures.     

Preservation of cell-based immunotherapies for clinical trials

In the unique supply chain of cellular therapies, preservation is important to keep the cell product viable. Many factors in cryopreservation affect the outcome of a cell therapy: (i) formulation and introduction of a freezing medium, (ii) cooling rate, (iii) storage conditions, (iv) thawing conditions and (v) post-thaw processing. This article surveys clinical trials of cellular immunotherapy that used cryopreserved regulatory, chimeric antigen receptor or gamma delta T cells, dendritic cells or natural killer (NK) cells. Several observations are summarized from the given information. The aforementioned cell types have been similarly frozen in media containing 5–10% dimethyl sulfoxide (DMSO) with plasma, serum or human serum albumin. Two common freezing methods are an insulated freezing container such as Nalgene Mr. Frosty and a controlled-rate freezer at a cooling rate of -1°C/min. Water baths at approximately 37°C have been commonly used for thawing. Post-thaw processing of cryopreserved cells varied greatly: some studies infused the cells immediately upon thawing; some diluted the cells in a carrier solution of varying formulation before infusion; some washed cells to remove cryoprotective agents; and others re-cultured cells to recover cell viability or functionality lost due to cryopreservation. Emerging approaches to preserving cellular immunotherapies are also described. DMSO-free formulations of the freezing media have demonstrated improved preservation of cell viability in T lymphocytes and of cytotoxic function in natural killer cells. Saccharides are a common type of molecule used as an alternative cryoprotective agent to DMSO. Improving methods of preservation will be critical to growth in the clinical use of cellular immunotherapies.     

Deep-supercooling for extended preservation of adipose-derived stem cells

Cell preservation is an enabling technology for widespread distribution and applications of mammalian cells. Traditional cryopreservation via slow-freezing or vitrification provides long-term storage but requires cytotoxic cryoprotectants (CPA) and tedious CPA loading/unloading, cooling, and recovering procedures. Hypothermic storage around 0–4 °C is an alternative method but only works for a short period due to its high storage temperatures. Here, we report on the deep-supercooling (DSC) preservation of human adipose-derived stem cells at deep subzero temperatures without freezing for extended storage. Enabled by surface sealing with an immiscible oil phase, cell suspension can be preserved in a liquid state at −13 °C and −16 °C for 7 days with high cell viability, retention of stemness, attachment, and multilineage differentiation capacities. These results demonstrate that DSC is an improved short-term preservation approach to provide off-the-shelf cell sources for booming cell-based medicine and bioengineering.