 |
|||
|
ABSTRACT ARCHIVE |
|||
|
|||
|
|
 |
|
|
|
CryoLetters is a bimonthly, international journal for low temperature science and technology |
|
|
|
CryoLetters 36 (4), 226-236 (2015) LONG-TERM PRECONDITIONING OF PLANTLETS: A PRACTICAL METHOD FOR ENHANCING SURVIVAL OF PINEAPPLE (Ananas comosus (L.) Merr.) SHOOT TIPS CRYOPRESERVED USING VITRIFICATION Wei-Hsin Hu1, 3, Shu-Fen Liu2, 3 and Song-Iuan Liaw2* 1Department of Biology, National Museum of Natural Science, Taichung, Taiwan Abstract BACKGROUND: The purpose of this study was to develop an efficient cryopreservation protocol for pineapple (Ananas comosus (L.) Merr.) shoot tips. OBJECTIVE: The optimal state of pineapple plantlets was investigated by using sucrose preconditioning to enhance survival after cryostorage. METHODS: To achieve a suitable state of plantlets before cryopreservation, 0.2 M to 0.4 M sucrose concentrations combined with short- (0–7 days), medium- (15–30 days), and long-term (75–150 days) preconditioning periods were compared. RESULTS AND CONCLUSION: The highest survival (100%) was achieved using the following procedure: intact plantlets underwent long-term preconditioning with 0.2 M sucrose for 135 days, dissected shoot tips were treated with a loading solution containing 2.0 M glycerol + 0.4 M sucrose for 60 min at 25°C and the shoot tips were dehydrated in PVS2 for 2 h at 0°C before being plunged in liquid nitrogen. Rewarming was conducted in a water-bath for 30 s at 40°C and PVS2 was replaced with a 1.2 M sucrose solution for 30 min at 25°C. The shoot tips were transferred on semisolid medium and left in the dark for 1 week, then in dim light for 3 weeks. Keywords: sucrose preconditioning, pineapple, vitrification
CryoLetters 36 (4), 237-242 (2015) COLD HARDINESS AND RANGE OF THE MYRIAPOD Angarozonium amurense (POLYZONIIDAE, DIPLOPODA, ARTHROPODA) IN PERMAFROST ENVIRONMENTS Daniil I. Berman1, Ekaterina N. Meshcheryakova1* 1Laboratory of Biocenology, Institute of Biological Problems (North, Far East Branch), Russian Academy of Sciences, Magadan, Russia; Abstract BACKGROUND: Angarozonium amurense (Gerstfeldt, 1859) is the only one out of more than a hundred diplopod species described in Siberia and the Far East that inhabits regions with solid permafrost. OBJECTIVE: To evaluate the cold hardiness of A. amurense that allows this species to inhabit permafrost regions. METHODS: The survival temperature thresholds and supercooling points (SCP) were measured. RESULTS: The temperature thresholds for adult animals’ survival are 8.5°C in summer and 27°C in winter. Average SCP decreases from 7.7±0.3°С in summer to 16.9±0.5°С in winter. Water content decreases from 55.7±1.9°% in summer to 49.4±1.6% in winter. CONCLUSION: The cold hardiness of A. amurense sets the record in this class of animals. It allows it to overwinter in the upper 15 centimeters layer of soil in most biotopes of the coldest permafrost regions in North Asia. Keywords: Diplopoda, Angarozonium amurense, cold hardiness, permafrost.
CryoLetters 36 (4), 243-251 (2015) THE COMBINED USE OF HONEY, GARLIC (ALLIUM SATIVUM L.) AND SKIMMED MILK AS AN EXTENDER FOR CHILLING SHEEP SEMEN Rodrigo Jerez-Ebensperger1*, Lydia Gil1, Noelia Gonzalez1 1Department of Animal Pathology, Obstetrics and Reproduction Area, Faculty of Veterinary Medicine; Abstract BACKGROUND: Sugars are the energetic source for sperm to maintain the metabolic process, and the antibiotics slow down sperm degradation. OBJECTIVE: To study the effects of rosemary honey as energy source and cryoprotectant in combination with garlic as a natural antibiotic on the quality of ram spermatozoa upon cooling. MATERIALS AND METHODS: The ejaculates from three rams were evaluated at different times during cooling to determine its post-dilution quality. RESULTS: Glycerol and dimethylformamide in conjunction with honey and garlic significantly improve the survival of spermatozoa. CONCLUSION: The addition of honey and garlic reduces sperm deterioration when stored at 4°C. Keywords: Acridine orange; spermatozoa, ram, garlic, honey, OxyDNA, skim milk.
CryoLetters 36 (4), 252-263 (2015) physiological, Cytological and BIOCHEMICAL STABILITY of Medicago sativa L. cell culture after 27 years of cryogenic storage Liudmila A. Volkova1, Valentina V. Urmantseva1, 1Department of Cell Biology and Biotechnology, Timiryazev Institute of Plant Physiology of Russian Academy of Sciences, Botanicheskaya ul. 35, Moscow 127276, Russia. Abstract BACKGROUND: The efficiency of long-term cryogenic storage to prevent somaclonal variations in plant cell cultures and retain their major cytogenetic and biochemical traits remains under debate. In particular, it is not clear how stress conditions associated with cryopreservation, such as low temperature, dehydration and toxic action of some cryoprotectants (DMSO in particular), affect post-storage regrowth and genetic integrity of cell samples. OBJECTIVE: We assessed growth, cytogenetic and biochemical characteristics of the peroxidase-producing strain of Medicago sativa L. cell culture recovered after 27 years of cryogenic storage as compared to the same culture before cryopreservation. MATERIALS AND METHODS: In 1984, M. sativa L. cell culture was cryopreserved using programmed freezing and 7% DMSO as a cryoprotectant. In 2011, after rewarming in a water bath at 40°С for 90 s, cell culture was recovered and proliferated. Viability, growth profile, mitotic index, ploidy level, peroxidase activity and cell response to hypothermia and osmotic stress were compared between the recovered and the initial cell cultures using the records available from 1984. RESULTS: Viability of alfalfa cell culture after rewarming was below 20% but it increased to 80% by the 27th subculture cycle. Recovered culture showed higher mitotic activity and increased number of haploid and diploid cells compared to the initial cell line. Both peroxidase activity and response to abiotic stress in the recovered cell culture were similar to that of the initial culture. CONCLUSION: Cryopreservation by programmed freezing was effective at retaining the main characteristics of M. sativa undifferentiated cell culture after 27 years of storage. According to available data, this is longest period of successful cryopreservation of plant cell cultures reported so far. After storage, there was no evidence that DMSO had any detrimental effect on cell viability, growth or cytogenetics. Keywords: alfalfa, cell culture, cryopreservation, peroxidase activity, cytogenetic stability, long-term storage, mitotic index, proliferation.
CryoLetters 36 (4), 264-269 (2015) EFFECTS OF ORVUS ES PASTE ON THE MOTILITY AND VIABILITY OF YAK (BOS GRUNNIENS) EPIDIDYMAL AND EJACULATED SPERMATOZOA AFTER FREEZING AND THAWING Megumi Shimazaki1, Rentsenkhand Sambuu2, Yoko Sato1, 1Department of Animal Reproduction, Joint Faculty of Veterinary Medicine, Yamaguchi University, Yamaguchi 753-8515, Japan Abstract BACKGROUND: The addition of the detergent Orvus ES Paste (OEP) to semen freezing extenders has been observed to improve the post-thaw survival and longevity of spermatozoa from various species but has never been evaluated for yak spermatozoa. OBJECTIVE: This study evaluated the effects of OEP on the post-thaw motility and viability of epididymal and ejaculated yak spermatozoa. MATERIALS AND METHODS: Semen samples were frozen and thawed in semen freezing extender supplemented with 0%, 0.375%, 0.75% or 1.5% OEP. The motility and viability of frozen-thawed spermatozoa were evaluated before and after 3 h of incubation. RESULTS: The addition of 0.75% OEP to the freezing extender significantly improved the mean motility and viability values of both the epididymal and ejaculated spermatozoa immediately after thawing, but the beneficial effects on motility disappeared after 3 h of incubation. CONCLUSION: Our findings indicate that the addition of 0.75% OEP is effective for the preservation of yak spermatozoa. Keywords: Bos grunniens, cryopreservation, detergent, freezing extender, spermatozoa
CryoLetters 36 (4), 270-277 (2015) RELATIONSHIP BETWEEN SUPERCOOLING CAPABILITY AND CRYOPROTECTANT CONTENT IN EGGS OF PARARCYPTERA MICROPTERA MERIDIONALIS (ORTHOPTERA: ACRYPTERIDAE) Xiao-Rong Zhou#, Yan-Yan Li#, Na Li and Bao-Ping Pang* Research Center for Grassland Entomology, Inner Mongolia Agricultural University, Hohhot, China. Abstract BACKGROUND: Grasshoppers are major agricultural pests throughout the world. The egg stage is important for the low temperature resistance, and almost all grasshoppers overwinter in the egg stage. OBJECTIVE: To study the relationship between cold hardiness and cryoprotectant content in Pararcyptera microptera meridionalis eggs. MATERIALS AND METHODS: The supercooling point (SCP) of the eggs was measured, along with the contents of water, fat, amino acids, low molecular sugars and polyols. RESULTS: SCP, water content and glucose concentration decreased during egg development, whereas the contents of fat, trehalose, glycerol, inositol and sorbitol increased. SCP is negatively correlated with the concentrations of fat, trehalose, glycerol, inositol and sorbitol, but positively with water content and glucose concentration. Among low molecular weight sugars and polyols tested in eggs, trehalose concentration was highest, followed by glycerol. Although total content of free amino acids did not change much, of the tested 17 free amino acids in eggs, proline and glutamine had increased by 46.3% and 13.2%, respectively, and both showed a negative correlation with SCP. Stepwise regression analysis showed that proline, glycerol, trehalose and inositol contribute most to the SCP depression. Cold acclimation at 0C increased the contents of trehalose and glycerol, and decreased SCP. CONCLUSION: The increase of the supercooling capacity in P. microptera meridionalis eggs during development could be attributed mainly to proline, glycerol, trehalose and inositol. Cold acclimation enhances supercooling capacity via glycerol and trehalose. Keywords: Pararcyptera microptera meridionalis, supercooling capacity, cold acclimation, low molecular weight carbohydrates, amino acids.
CryoLetters 36 (4), 278-284 (2015) TEMPORARY STORAGE OF BOVINE SEMEN CRYOPRESERVED IN LIQUID NITROGEN ON DRY ICE AND REFREEZING OF FROZEN-THAWED SEMEN A.M. Abdussamad1, M. Gauly2 and W. Holtz* Department of Animal Science, Georg-August-University Goettingen,Albrecht-Thaer-Weg 3, 37075 Goettingen, Germany Abstract Two experiments were conducted. The purpose of Experiment 1 was to investigate whether viability of bovine semen stored in liquid nitrogen (-196°C) will be adversely affected by temporary exposure to dry ice (-79°C). It was convincingly shown that post thaw-motility was not affected, regardless whether semen was thawed immediately or after being returned to liquid nitrogen. Shipping or temporary storage on dry ice, thus, is a viable option. In Experiment 2, refreezing of frozen-thawed semen was attempted. The proportion of motile spermatozoa was reduced by a factor of ten to between 6.0% and 7.4%, regardless whether thawing occurred directly after removal from liquid nitrogen or after an interim period on dry ice. When semen was refrozen on dry ice before being returned to liquid nitrogen, motility rates were significantly improved (13.0% to 17.0%, P<0.05). In both experiments sperm cells that remained motile displayed vigorous forward movement and normal morphological appearance. Keywords: Cryopreservation; Semen; Cattle; Liquid nitrogen; Dry ice
CryoLetters 36 (4), 285-288 (2015) DETERMINATION OF CONVECTIVE HEAT TRANSFER COEFFICIENT AT THE OUTER SURFACE OF A CRYOVIAL BEING PLUNGED INTO LIQUID NITROGEN Tao Wang1, Gang Zhao1,2*, Heyu Tang1 and Zhendong Jiang1 1Centre for Biomedical Engineering, Department of Electronic Science & Technology, University of Science and Technology of China, Hefei; Abstract BACKGROUND: Cell survival upon cryopreservation is affected by the cooling rate. However, it is difficult to model the heat transfer process or to predict the cooling curve of a cryoprotective agent (CPA) solution due to the uncertainty of its convective heat transfer coefficient (h). OBJECTIVE: To measure the h and to better understand the heat transfer process of cryovials filled with CPA solution being plunged in liquid nitrogen. MATERIALS AND METHODS: The temperatures at three locations of the CPA solution in a cryovial were measured. Different h values were selected after the cooling process was modeled as natural convection heat transfer, the film boiling and the nucleate boiling, respectively. And the temperatures of the selected points are simulated based on the selected h values. h was determined when the simulated temperature best fitted the experimental temperature. RESULTS: When the experimental results were best fitted, according to natural convection heat transfer model, h1 = 120 W/(m2·K) while due to film boiling and nucleate boiling regimes hf = 5 W/(m2·K) followed by hn = 245 W/(m2·K). These values were verified by the differential cooling rates at the three locations of a cryovial. CONCLUSION: The heat transfer process during cooling in liquid nitrogen is better modeled as film boiling followed by nucleate boiling. Keywords: Cryoprotective agent, Cryovial, Convective heat transfer coefficient, Cooling rate, Finite element method |
|
|
|
|
|
|
|
|
|
|||||||||||||||||||||||
|
Abstracts |
|||||||||||||||||||||||||||
|
|||||||||||||||||||||||||||
|
For Abstracts published from meetings, such as SLTB meetings, go to the
relevant Volume Year of the journal (above). For Full text Free Access Content (from 2000 onwards) go to CryoLetters at Ingenta and look for the blue symbol. |
|||||||||||||||||||||||||||
