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Abstracts: CryoLetters 39 (6), 2018

 

 

Volume 39, No. 6 November/December 2018

ISSN 0143-2044

 

 


An improved cryopreservation protocol for Mentha spp. Based on PVS3 as the cryoprotectant
Angelika Senula, Doris Büchner, E.R. Joachim Keller
and Manuela Nagel

345-353

 

 


Effects of long-term storage on some spermatological parameters in cryopreserved bull semen
Numan Akyol, Ömer Varışlı and Sedat Hamdi Kızıl

354-358

 

 


The assessment of cryopreservation on the quality of endangered Oravka rooster spermatozoa using casa and cytometry
Andrea Svoradová, Lenka Kuželová, Jaromír Vašíček,
Lucia Olexíková, Andrej Baláži, Barbora Kulíková, Cyril Hrnčár,
Alexander Ostro, Marek Bednarczyk and Peter Chrenek

359-365

 

 


Field performance of cryopreserved seed-derived maize plants
Melissa Arguedas, Ariel Villalobos, Daviel Gómez, Lázaro Hernández, Byron E. Zevallos, Inaudis Cejas, Lourdes Yabor,
Marcos Edel Martínez-Montero and José Carlos Lorenzo

366-370

 

 


Reduced glutathione and ATP in the seminal cryopreservation of tambaqui
Fernanda Alves Pereira, Carine Dahl Corcini,
Alessandra Cardoso da Silva,Diego Martins Pires,
Jéssica Ribeiro Pereira, Stela Mari Meneghello Gheller,
Danilo Pedro Streit Junior, Alexandre Nizio Maria
and Antonio Sergio Varela Junior

371-379

 

 


Theoretical estimation of the optimum cooling rate of a cell suspension at linear freezing modes based on a two factor theory of cryodamage
O.I. Gordiyenko, S.Ye Kovalenko., I.F. Kovalenko,
V.V. Ogurtsova and A.F. Todrin

380-385

 

 


Effect of cholesterol-loaded cyclodextrin on membrane and acrosome status of hariana bull sperm during cryopreservation
Hanuman P Yadav, A Kumar, N Shah, DS Chauhan, SA Lone, DK Swain and A Saxena

386-390

 

 


Hypothermic preservation of red blood cells in different conditions of inert gas xenon: hyperbaria and clathrates
Alexander Ponomarev, Victor V. Rodin, Leonid Gurevich,
Vsevolod Melekhin and Oleg Makeev

391-400

 

 


Freezing of stallion semen: in vitro and in vivo evaluation of sperm motility and acrosin activity in frozen-thawed semen with addition of post-diluent extenders
Felipe Augusto Boudoux Martins Sales, José Carlos
Ferreira-Silva, Joane Isis Travassos Vieira, Sarah Romini
Lima Basto, Paulo Castelo Branco Gouveia Filho,
Jorge Motta Rocha, Marcelo Tigre Moura
and Marcos Antonio Lemos Oliveira

401-407

 

 


Authors Index

Keywords and Editorial Announcement

408-410

411-413

 

 

 

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CryoLetters 39 (6), 345-353 (2018)
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AN IMPROVED CRYOPRESERVATION PROTOCOL FOR Mentha spp. BASED ON PVS3 AS THE CRYOPROTECTANT

Angelika Senula, Doris Büchner, E.R. Joachim Keller and Manuela Nagel*

Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstraße 3, D-06466 Seeland OT Gatersleben, Germany
*Corresponding author email:

Abstract

BACKGROUND: Vitrification approaches are widely used to cryopreserve Mentha spp. genetic resources. OBJECTIVE: Here, we compared the response of 20 different Mentha species and hybrids during cryopreservation and elucidated the efficacy of two cryoprotectants. MATERIALS AND METHODS: One hundred and fifty three Mentha spp. accessions were cryopreserved using in vitro plants maintained under slow-growth storage and PVS2 or PVS3 as cryoprotectants. RESULTS: The cryoprotectant PVS2 was effective for all species, except M. requienii and M. villosanervata. The use of PVS3 increased the proportion of explants able to regrow after rewarming. The outbreak of endophytes upon rewarming was both less frequent and less severe when PVS3 replaced PVS2. CONCLUSION: Both PVS2 and PVS3 can be used as cryoprotectant for all the species and accessions of Mentha spp. surveyed. Since higher regenerations were achieved using PVS3, and since the risk of an endophyte outbreak was reduced, this cryoprotectant should be preferred in future for cryopreserving Mentha spp.

Keywords: gene bank, glass, mint, PVS2, regrowth, endophyte

 

 

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CryoLetters 39 (6), 354-358 (2018)
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Effects of long-term storage on some spermatological parameters in cryopreserved bull semen

Numan Akyol1, Ömer Varışlı1* and Sedat Hamdi Kızıl2

1Department of Reproduction & Artificial Insemination, Faculty of Veterinary Medicine, Kırıkkale University, Kırıkkale;
2International Livestock Research and Training Center Ankara, Turkey.
*Corresponding author email:
omer.dvm@gmail.com

Abstract

BACKGROUND: The effect of the long-term storage in liquid nitrogen on semen quality has not be reported. OBJECTİVE: The study measured the spermatological parameters of bull sperm after the long-term and short-term storage. MATERIALS AND METHODS: Vintage semen (obtained from 5 Brown Swiss bulls and frozen 30 years ago) and newly frozen semen collected from 5 bulls of the same breed and prepared at the International Center for Livestock Research and Training were used. For each bull, 10 straws (0.25 ml) were thawed and pooled. Sperm samples were analyzed by flow cytometry, computer assisted sperm analysis, and total oxidant-antioxidant levels were also tested. RESULTS: The ratios of necrotic (P < 0.001) and apoptotic (P = 0.006) spermatozoa, and the ratios of total antioxidant status (TAS) and total oxidant status (TOS) were significantly higher (P < 0.001) in long-term frozen spermatozoa. However, the early necrotic ratios (P < 0.001), velocity average pathway (VAP) (P = 0.008) and velocity curvi linear (VCL) (P = 0.01) values of long-term frozen semen were lower compared with short-term frozen semen. While necrotic and apoptotic spermatozoa ratios and total oxidant level were higher, VAP/VCL ratios were lower in long-term frozen sperms compared to short-term frozen semen. CONCLUSION: Long-term storage of sperm may adversely affect the spermatological and oxidative parameters of Brown Swiss bull spermatozoa.

Keywords: Bull semen, long-term storage, cryopreservation, Brown Swiss.

 

 

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CryoLetters 39 (6), 359-365 (2018)
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THE ASSESSMENT OF CRYOPRESERVATION ON THE QUALITY OF ENDANGERED ORAVKA ROOSTER SPERMATOZOA USING CASA AND CYTOMETRY

Andrea Svoradová*1, Lenka Kuželová2, Jaromír Vašíček3, 7,
Lucia Olexíková3, Andrej Baláži3, Barbora Kulíková3, Cyril Hrnčár4,
Alexander Ostro5, Marek Bednarczyk6 and Peter Chrenek3, 7

1Constantine the Philosopher University in Nitra, Faculty of Natural Sciences, Nitra;
2Slovak University of Agriculture in Nitra, Research Centre AgroBioTech, Nitra;
3National Agricultural & Food Centre, Research Institute for Animal Production Nitra, Lužianky;
4Slovak University of Agriculture in Nitra, Faculty of Agrobiology & Food Resources, Nitra;
5Pavol Jozef Šafárik University in Košice, Faculty of Medicine, Košice;
6
Department of Animal Biochemistry and Biotechnology, UTP University of Science and Technology, Bydgoszcz, Poland,
7Slovak University of Agriculture in Nitra, Faculty of Biotechnology & Food Sciences, Nitra, Slovak Republic.
*Corresponding author email:
svoradovaandreal@gmail.com

Abstract

BACKGROUND: Preservation of genetic resources in gene bank is necessary for conservation of endangered poultry species. OBJECTIVE: This study is to characterize Oravka rooster semen quality. MATERIALS AND METHODS: Heterospermic pool was diluted (1:1 by volume) in a freeze medium composed of a commercial diluent (Kobidil+, K), 8% dimethyl sulphoxide (DMSO) or 8% ethylene glycol (EG) or 8% glycerol (GL), and then frozen in liquid nitrogen vapour. RESULTS: Spermatozoa in the GL/K+ had significantly higher number of motile and progressively moving spermatozoa (p<0.05) than in DMSO/K+ and EG/K+ groups. The percentage of apoptotic and necrotic spermatozoa were significantly higher in the DMSO/K+ and EG/K+ groups compared with the GL/K+ group. Based on the total motility, progressive movement parameters and viability, our study showed that 8% GL diluted in Kobidil+ provided the highest cryoprotective effect on the Oravka rooster spermatozoa.

Keywords: Oravka breed, poultry, rooster spermatozoa, CASA, flow cytometry.

 

 

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CryoLetters 39 (6), 366-370 (2018)
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FIELD PERFORMANCE OF CRYOPRESERVED SEED-DERIVED MAIZE PLANTS

Melissa Arguedas1, Ariel Villalobos1, Daviel Gómez1,
Lázaro Hernández1, Byron E. Zevallos2, Inaudis Cejas1,
Lourdes Yabor1, Marcos Edel Martínez-Montero1
and José Carlos Lorenzo1*

1 Laboratory for Plant Breeding and Conservation of Genetic Resources, Centro de Bioplantas, Universidad de Ciego de Ávila, Ciego de Ávila 69450, Cuba
2 Escuela Superior Politécnica Agropecuaria de Manabí Manuel Félix López (ESPAMMFL), Campus Politécnico El Limón, Carrera de Ingeniería Agrícola, Calceta, Manabí, Ecuador.
*Corresponding author email:
jclorenzo@bioplantas.cu URL: www.bioplantas.cu.

Abstract

BACKGROUND: Cryo-preservation of plant materials in liquid nitrogen (LN) has been described as a suitable technology to conserve genetic resources of several species. However, the potential effects of LN in the subsequent plant growth in the field should be studied before large-scale implementation of cryopreserved germplasm banks. OBJECTIVE: To describe the field performance of cryopreserved seed – derived maize adult plants. MATERIALS AND METHODS: Germination percentage and numbers of leaves and ears per plant, internodes in stems, middle - aged leaf length, plant height, ear traits and weight of 100 seeds were recorded. RESULTS: Statistically significant differences between adult plants derived from cryopreserved seeds and the control treatment were not observed (t-test, p=0.05).  CONCLUSION: The results presented confirm at the phenotype level the effectiveness of maize seed cryostorage to preserve and regenerate true-to-type plants.

Keywords: germplasm preservation; liquid nitrogen; Zea mays L.

 

 

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CryoLetters 39 (6), 371-379 (2018)
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REDUCED GLUTATHIONE AND ATP IN THE SEMINAL CRYOPRESERVATION OF TAMBAQUI

Fernanda Alves Pereira1, Carine Dahl Corcini2, 3, *,
Alessandra Cardoso da Silva4, Diego Martins Pires4,
Jéssica Ribeiro Pereira1, Stela Mari Meneghello Gheller4,
Danilo Pedro Streit Junior5, Alexandre Nizio Maria6
and Antonio Sergio Varela Junior2, 3

1Programa de pós graduação em Biologia de Ambientes Aquáticos Continentais, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande, Rio Grande, RS, Brasil
2Reprodução Animal Comparada- RAC, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande, Rio Grande, RS, Brasil
3ReproPel, Faculdade de Veterinária, Universidade Federal de Pelotas, Pelotas, RS, Brasil
4Programa de pós graduação em Veterinária, Faculdade de Veterinária, Universidade Federal de Pelotas, Pelotas, RS, Brasil.
5Aquam, Departamento de Zootecnia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil
6Embrapa Tabuleiros Costeiros, Aracaju, SE, Brasil
*Corresponding author email: corcinicd@gmail.com

Abstract

BACKGROUND: The aim of this study was to analyze the effect of reduced glutathione (GR) and adenosine triphosphate (ATP) in cryopreserved sperm tambaqui, Colossoma macropomum.. OBJECTIVE: Semen samples were frozen at different concentrations of GR (2, 4, and 6 mM) and ATP (7.5, 15, 30 mM) evaluated separately. MATERIALS AND METHODS: Different concentrations of GR and ATP were added to the diluent Beltsville Thawing Solution 10 % of DMSO (extender control). The thawed sperm was evaluated as the kinetic parameters for Computer Assisted Semen Analysis (CASA) and cell feature parameters by flow cytometry. RESULTS: All concentrations of GR and ATP had gradual reduction in the production of reactive oxygen species, mainly ATP 15; 22.5 mM and 30 mM, which decreased by 50% or more when compared to the control. All concentrations of GR and ATP did not change (P> 0.05) DNA fragmentation and lipid peroxidation. The membrane fluidity GR in concentrations 4 and 6 mM were better than control; and the concentrations of 2 and 4 mM maintained more efficiently to motility period, total and progressive motility. CONCLUSION: Thus, it is indicated the use of BTS extender with 10% DMSO added 4 mM GR thawed sperm Tambaqui C. macropomum.

Keywords: Antioxidant; conservation; sperm; fish; ROS

 

 

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CryoLetters 39 (6), 380-385 (2018)
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THEORETICAL ESTIMATION OF THE OPTIMUM COOLING RATE OF A CELL SUSPENSION AT LINEAR FREEZING MODES BASED ON A TWO FACTOR THEORY OF CRYODAMAGE

O.I. Gordiyenko*, S.Ye Kovalenko., I.F. Kovalenko, V.V. Ogurtsova
and A.F. Todrin

Institute for Problems of Cryobiology and Cryomedicine NAS of Ukraine, Kharkiv, Ukraine
*Corresponding author email:
olga.gordiyenko.ipcic@gmail.com

Abstract

BACKGROUND: According to the two-factor theory cryodamage arises from intracellular crystallization and solution effects due to freeze concentration. OBJECTIVE: The study aims to evaluate the contribution of two types of cryodamages that are related to extra- and intracellular crystallization. METHODS: The probability of intracellular crystal formation during cooling of cell suspension in cryoprotective solution has been determined based on general thermodynamics theory. RESULTS: According to the obtained correlations and taking into account of the individual characteristics of yeast cells and murine enterocytes, the optimal cooling rates during freezing of these cells in cryoprotectant solutions were determined. CONCLUSION: The proposed algorithm for the estimation of the optimal cooling rates at linear freezing mode of a particular cellular suspension can be utilized to develope methods for cryopreservation of different cell suspensions.

Keywords: cryodamage, cryopreservation, freeze concentration, intracellular crystallization, murine enterocytes, Saccharomyces cerevisiae.

 

 

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CryoLetters 39 (6), 386-390 (2018)
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EFFECT OF CHOLESTEROL-LOADED CYCLODEXTRIN ON MEMBRANE AND ACROSOME STATUS OF HARIANA BULL SPERM DURING CRYOPRESERVATION

Hanuman P Yadav1, A Kumar2, N Shah1, DS Chauhan3, SA Lone1,
DK Swain4 and A Saxena2

1Artificial Breeding Research Center (ABRC), Animal Reproduction, Gynecology and Obstetrics, ICAR-National Dairy Research Institute, Haryana;
2Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, U.P. Pandit Deendayal Upadhayaya Pashu Chikitsa Vigyan Vishwavidyalaya Evam Go Anusandhan Sansthan (DUVASU), Uttar Pradesh;
3College of Biotechnology, (DUVASU), Uttar Pradesh; 4Department of Veterinary Physiology, College of Veterinary Science and Animal Husbandry, (DUVASU), Uttar Pradesh, India.
*Corresponding author email:
drhanumanndri@gmail.com

Abstract

BACKGROUND: The membrane and acrosomal integrity of sperm play a vital role in fertilization process; however they are compromised upon cryopreservation. OBJECTIVE: To study the effect of cholesterol-loaded cyclodextrin (CLC) on membrane and acrosome status of Hariana bull sperm during cryopreservation. MATERIALS AND METHODS: Semen samples collected from Hariana bulls with mass motility ≥ 3+ and individual progressive motility ≥ 70% were utilized in the study. Each ejaculate was split into two parts, one part being evaluated freshly for various seminal attributes and the other part being diluted in Tris diluent (without egg yolk and glycerol) to obtain a final concentration of 120×106 sperm/mL. The diluted semen was divided into four treatments: Group I, without CLC (control); Group II, with CLC at 0.5 mg per 120 million sperm; Group III, at 1.0 mg per 120 million sperm; Group IV, at 2.0 mg per 120 million sperm. All aliquots were incubated for 15 min at 37°C and each sample was diluted with Egg yolk-Tris-Glycerol (EYTG) extender up to 80×106 sperm/mL. The diluted semen samples were packed in French mini straws (0.25 mL), sealed and equilibrated at 4°C for 4 h followed by cryopreservation. The samples at pre-freeze and post-thaw stage were evaluated for membrane and acrosomal integrity, as well as primary, secondary and tertiary acrosomal damages. RESULTS: The membrane and acrosomal integrity was significantly higher in group II as compared to groups I, III, and IV, at pre-freeze and post-thaw stage (P<0.05). The primary and secondary acrosomal damage were significantly reduced in group II compared to other groups (P<0.05). No significant difference in tertiary acrosomal damage was found among different groups. CONCLUSION: CLC improves the membrane and acrosomal integrity, and reduces primary and secondary acrosomal damages during cryopreservation of Hariana bull sperm.

Keywords: bull sperm, cholesterol-loaded cyclodextrin, cryopreservation

 

 

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CryoLetters 39 (6), 391-400 (2018)
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HYPOTHERMIC PRESERVATION OF RED BLOOD CELLS IN DIFFERENT CONDITIONS OF INERT GAS XENON: HYPERBARIA AND CLATHRATES

Alexander Ponomarev1, 2, 3, Victor V. Rodin4, 5*, Leonid Gurevich6,
Vsevolod Melekhin1, 3 and Oleg Makeev1, 3

1Dept. of Molecular and Cell Technologies, Ural State Medical University, 620028 Ekaterinburg, Russia. 
2Regional Children's Hospital №1, 620149 Ekaterinburg, Russia.
3Institute for Medical Cell Technologies, 620026 Ekaterinburg, Russia.
4Institute of Organic Chemistry, Johannes Kepler University Linz, Altenbergerstraße 69, 4040 Linz, Austria.
5Glasgow Experimental MRI Centre, Institute of Neuroscience and Psychology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, G61 1QH, United Kingdom.
6Department of Materials and Production, Aalborg University, Aalborg, Denmark
*Corresponding author e-mail:
victor.rodin@jku.at

Abstract

BACKGROUND: Xenon is an inert gas promising for the preservation of biomaterials, which forms clathrate hydrates above 0°C.  OBJECTIVE: This study addresses the effect of hyperbaric xenon (P = 303 kPa) and water-xenon clathrates (P ≥ 608 kPa) on 30 days preservation of red blood cells (RBCs) at +4°C.  MATERIALS AND METHODS: RBCs from healthy human donors were preserved under four different conditions: without preservatives (negative control), in CPDA-1, hyperbaric xenon, and xenon clathrate hydrates. RESULTS: The qualitative (mean RBC volume, anisocytosis degree and mean osmotic fragility) and quantitative characteristics (RBC count and hemolysis degree) of preserved RBCs were measured. CONCLUSION: The positive role of hyperbaric xenon in the preservation of erythrocytes is attributed to the equilibrium extraction and displacement of O2 and CO2 by xenon. The effect is presumably due to a lowering of oxygen concentration and a decrease in the production of reactive oxygen species.

Keywords: xenon, inert gases, blood preservation, hyperbaric xenon, xenon clathrates

 

 

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CryoLetters 39 (6), 401-407 (2018)
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FREEZING OF STALLION SEMEN: IN VITRO AND IN VIVO EVALUATION OF SPERM MOTILITY AND ACROSIN ACTIVITY IN FROZEN-THAWED SEMEN WITH ADDITION OF POST-DILUENT EXTENDERS

Felipe Augusto Boudoux Martins Sales1, José Carlos Ferreira-Silva1,
Joane Isis Travassos Vieira1, Sarah Romini Lima Basto,
Paulo Castelo Branco Gouveia Filho1, Jorge Motta Rocha2,
Marcelo Tigre Moura1 and Marcos Antonio Lemos Oliveira1*

1 Laboratório de Biotécnicas Reprodutivas, Universidade Federal Rural de Pernambuco, Recife-PE, Brasil.
2 Universidade Federal do Rio Grande do Norte, Natal-RN, Brasil.
*Corresponding author email:
maloufrpe@gmail.com

Abstract

BACKGROUND: Post-diluents could potentially increase semen cryotolerance, but remain poorly explored in horses. OBJECTIVE: The aim was to evaluate the efficiency of post-diluents on frozen-thawed semen viability of two stallions (S1-S2). MATERIALS AND METHODS: The cryopreserved semen was thawed at 50°C for 40 seconds. Semen motility and acrosin activity (AA) were determined during the thermo-resistance test (TRT). RESULTS: progressive motility of S2 semen decreased after 60 and 90 minutes of TRT (TRT60 and TRT90) on the control compared to both post-diluents. The total motility of both S1 and S2 decreased on TRT60 and TRT90 semen control versus both Ringer and Merk post-diluents. The AA on S1 was higher than S2 throughout the TRT. Pregnancy rates after artificial insemination (AI) were similar among post-diluents and stallions. CONCLUSION: post-diluents do not contribute to predicting frozen-thawed semen fertility or the efficiency of equine AI.

Keywords: cryopreservation, thermo-resistance, motility, acrosin, fertility

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