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Abstracts: CryoLetters 42 (5), 2021

 

 

Volume 42 , No. 5 September/October 2021

ISSN 0143-2044

 

 


Perspective: Oocyte and embryo preservation in wild animals: an update (PDF)
G R Bhat and K A Sofi

251-260

 

 


Seasonal variation in heat shock proteins (HSP70 and HSP90) and their association with frozen semen quality and fertility in buffaloes
KH Parmar, V Singh, HH Savsani, FS Kavani, JS Rajoriya
and SA Lone

261-266

 

 


Cryopreservation of Borassus flabellifer L. (Arecaceae) using excised embryos: a first report for the genus Borassus
Kang Han, Ganesh K. Jaganathan, Lanlan He, Jiajin Li,
Yingying Han and Baolin Liu

267-271

 

 


The effect of glycosaminoglycans, extracted from the skin of tilapia, in the sperm freezing medium of Colossoma macropomum
Vanessa Alves Pereira, Renata Vieira do Nascimento,
 Priscila Silva de Almeida-Monteiro,
Mayara Setúbal Oliveira-Araújo, Yasmim Maia Ferreira,
Thais Maia Torres, Yara Silvino Sales,
Ianna Wivianne Fernandes Araújo,
José Ariévilo Gurgel Rodrigues, Johnny Peter Macedo Feitosa, Sandra de Aguiar Soares, Assis Rubens Montenegro
and Carminda Sandra Brito Salmito-Vanderley

272-282

 

 


Effects of cryolipolysis with adiposity plates located with the Crioplace™ concept in women
Patrícia Froes Meyer, Rodrigo Marcel Valentim da Silva,
Eneida de Morais Carreiro, Fábio dos Santos Borges,
Stephany Luanna Queiroga Farias,
Thamires Santos Machado Figueiredo
and Talita Duarte Martins

283-289

 

 


Ammonium-free medium is critical for regeneration of shoot tips of the endangered species Pogostemon yatabeanus cryopreserved using droplet-vitrification
Hyoeun Lee, Hana Park, Elena Popova, Young-Yi Lee,
Sang-Un Park and Haeng-Hoon Kim

290-299

 

 


Cryopreservation of a threatened medicinal plant, Valeriana jatamansi Jones, using vitrification and assessment of biosynthetic stability of regenerants
Shailika Sharma, Kiran Parasher, Papiya Mukherjee
and Yash Pal Sharma

300-308

 

 

 

 

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CryoLetters 42 (5), 251-260 (2021)
© CryoLetters,
editor@cryoletters.org

PERSPECTIVE:
OOCYTE AND EMBRYO PRESERVATION IN WILD ANIMALS: AN UPDATE

G R Bhat* and K A Sofi

Division of Veterinary Clinical Complex, Sher-e- Kashmir University of Agricultural Sciences and Technology of Kashmir
*Corresponding author's E-mail:
rasool_roshan0127@rediffmail.com

Abstract

The reduction in population genetic diversity due to inbreeding depression and the negative impact of human activity on habitats ultimately generates an extinction debt. Therefore, there is always a dire need to save wild population and to protect biodiversity. Preservation of wildlife female germplasm, i.e., oocytes and embryos, is a promising biotechnological tool to conserve species' biodiversity. Other applied tools of Assisted Reproductive Technology (ART) which assure conservation of endangered species include artificial insemination (AI), embryo transfer technology (ETT), and sperm cryopreservation. Only a few studies show the possibility of adapting the cryopreservation techniques developed for domestic animal female genetic material for use with wild animals. Difficulty is encountered in getting samples, accesses to animals for study, and the standardization of protocols for cryopreservation of such genetic material. Our meta-analysis of the literature (published or in press) and on-going studies found that biobanking for the preservation of vital tissues of wild animals is possible. Somatic tissue sections, ovarian tissues, sperms, oocytes and embryos are potential materials for preservation by vitrification. As vitrification is economical and easily applied, it appears to the best option currently available for the preservation of wildlife female genetics in order to conserve species' biodiversity.

Keywords: cryopreservation; embryo; oocyte; wild animals.

Download the paper: OOCYTE AND EMBRYO PRESERVATION IN WILD ANIMALS: AN UPDATE (PDF)

 

 

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CryoLetters 42 (5), 261-266 (2021)
© CryoLetters,
editor@cryoletters.org

SEASONAL VARIATION IN HEAT SHOCK PROTEINS (HSP70 AND HSP90) AND THEIR ASSOCIATION WITH FROZEN SEMEN QUALITY AND FERTILITY IN BUFFALOES

KH Parmar1, V Singh1, HH Savsani1, FS Kavani1, JS Rajoriya2 and SA Lone3*

1 Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, Junagadh Agricultural University, Junagadh-362001, Gujrat, India.
2 Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, Kuthulia, Rewa, 486001, India.
3 Intensive Cattle Development Centre, Chogal, Kupwara, 193221, Department of Animal Husbandry, Jammu and Kashmir, India.
*Corresponding author's E-mail:
drloneshabir@gmail.com

Abstract

BACKGROUND: Heat shock protein is considered as a potential indicator of animal adaptation to harsh environmental stresses and its expression has been correlated with resistance to stress. OBJECTIVE: To evaluate the seasonal variation in heat shock proteins (hsp70 and hsp90) and their association with frozen semen quality and fertility in buffaloes. MATERIALS AND METHODS: During summer and winter, ejaculates (n = 32) were collected from buffalo bulls, diluted with freshly prepared Andromed extender (maintained at 34 ºC) up to 80 million sperm per mL. The diluted semen was filled in French midi straws, equilibrated, and cryopreserved. The semen was evaluated at pre-freeze and post-thaw stages for heat shock proteins (HSP70 and HSP90), and sperm quality parameters. RESULTS: The levels of HSP70 were significantly (P = 0.00) higher in summer season compared to winter season. The HSP70 had a positive correlation with mass motility (P < 0.05; r = 0.39), live sperm count (P < 0.05; r = 0.47), and acrosomal integrity (P < 0.05; r = 0.37). The first artificial insemination conception rate (FAICR) had a positive correlation with HSP70 (P < 0.05; r = 0.47), and HSP90 (P < 0.05; r = 0.59), in frozen-thawed semen. CONCLUSION: The assessment of the levels of heat shock proteins may help in predicting cryo-tolerance and fertility of buffalo semen during various seasons.

Keywords: buffalo; bull; cryopreservation; fertility; HSPs; semen; sperm quality.

 

 

 

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CryoLetters 42 (5), 267-271 (2021)
© CryoLetters,
editor@cryoletters.org

CRYOPRESERVATION OF Borassus flabellifer L. (ARECACEAE) USING EXCISED EMBRYOS: A FIRST REPORT FOR THE GENUS Borassus

Kang Han1#, Ganesh K. Jaganathan1#, Lanlan He1, Jiajin Li1,
Yingying Han1 and Baolin Liu1

1Institute of Biothermal technology, University of Shanghai for Science and Technology, Shanghai 200093, P.R. China
Corresponding author's E-mail:
jganeshcbe@gmail.com
#Contributed equally

Abstract

BACKGROUND: Embryos of many palm species that produce desiccation-sensitive, i.e. recalcitrant seeds have been successfully cryopreserved. However, storage protocols for some genera such as Borassus are still lacking. MATERIALS AND METHODS: Mature fruits of Borassus flabellifer L were collected in south India at the time of natural dispersal (June-July). After removal of the pericarp and mesocarp, the pyrenes were dried in silica gel and the desiccation tolerance level determined. Isolated embryos were also subjected to drying, exposure to liquid nitrogen temperature and recovery in vitro. RESULTS: Mature fruits weighed an average mass of 0.9 kg and germinated to 89%. The moisture contents (MCs) of the pyrenes and embryos were 48 and 78%, respectively. Pyrenes dried in silica gel to c. 20% MC lost viability, whereas embryos could be dried to 11% MC with 55% survival. These results indicate that the seeds are relatively desiccation-sensitive. Embryos dried to 21 and 11% MC had 23 and 36% survival following exposure to liquid nitrogen for 48 h, respectively. CONCLUSION: There is a hydration level window between c. 10 and 20% MC that is optimal for the cryopreservation of Borassus embryos. However, long-term storage possibilities remain to be explored.

Keywords: desiccation-sensitivity; liquid nitrogen storage; palms; pyrene.

 

 

 

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CryoLetters 42 (5), 272-282 (2021)
© CryoLetters,
editor@cryoletters.org

THE EFFECT OF GLYCOSAMINOGLYCANS, EXTRACTED FROM THE SKIN OF TILAPIA, IN THE SPERM FREEZING MEDIUM OF Colossoma macropomum

Vanessa Alves Pereira1*, Renata Vieira do Nascimento1, Priscila Silva de Almeida-Monteiro1, Mayara Setúbal Oliveira-Araújo1, Yasmim Maia Ferreira1, Thais Maia Torres1, Yara Silvino Sales2, Ianna Wivianne Fernandes Araújo3, José Ariévilo Gurgel Rodrigues3, Johnny Peter Macedo Feitosa4, Sandra de Aguiar Soares4, Assis Rubens Montenegro1 and Carminda Sandra Brito Salmito-Vanderley1

1 Fish Reproduction Biotechnology Laboratory, Postgraduate Program in Veterinary Sciences, State University of Ceará, Fortaleza, Ceará, Brazil
2 Fish Reproduction Biotechnology Laboratory, State University of Ceará, Fortaleza, Ceará, Brazil
3 Researcher in Biotechnology of Aquatic Organisms, Federal University of Ceará, Fortaleza, Ceará, Brazil
4 Polymers and Material Innovation Laboratory, Federal University of Ceará, Fortaleza, Ceará, Brazil
 *Corresponding author's E-mail:
vanessabio35@gmail.com

Abstract

BACKGROUND: Sulfated polysaccharides from the skin of Nile tilapia (Oreochromis niloticus), added to the tambaqui (Colossoma macropomum) semen diluting medium, can be potential antioxidants and promote the maintenance of sperm quality. OBJECTIVE: To evaluate the use of different concentrations of glycosaminoglycans (GAGs) from the skin of Nile tilapia as a supplement in two cryogenic media for tambaqui semen. MATERIALS AND METHODS: Tambaqui males received a single dose of pituitary carp extract. The semen was collected for pool analysis and, later, cryopreserved in liquid nitrogen. The pools were diluted and frozen in a solution containing fish-specific powdered coconut water (ACP-104) and 10% DMSO or 5% Glucose and 10% DMSO and supplemented with different concentrations of GAGs. The controls had no GAGs addition. After 45 days, the samples were thawed by immersion in a water bath and evaluated for membrane and DNA integrity, morphology and sperm kinetics.  RESULTS: The parameters of linearity (LIN), straightness (STR) and DNA integrity of sperm frozen in 5% Glucose showed better results than ACP-104. For membrane integrity, concentrations of 0 and 1.0 mg/mL were better than 5 mg/mL. Semen motility in 5% Glucose showed superior results at concentrations lower than 5 mg/mL of GAGs. For VCL and VAP, in ACP-104, 3.0 mg/mL exceeded the other treatments. In 5% Glucose, for VCL, 4.0 mg/mL showed the lowest results compared to concentrations of 3.5 mg/mL and, for VAP, it also differed from 4.5 mg/mL CONCLUSION: Therefore, the skin of Nile tilapia has GAGs, in low concentrations, capable of improving the post-thawed sperm quality of tambaqui, especially in 5% Glucose medium.

Keywords: concentration; cryopreservation; diluent; fish sperm; reproduction; sulfated polysaccharide; Tambaqui.

 

 

 

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CryoLetters 42 (5), 283-289 (2021)
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editor@cryoletters.org

EFFECTS OF CRYOLIPOLYSIS WITH ADIPOSITY PLATES LOCATED WITH THE CRIOPLACE™ CONCEPT IN WOMEN

Patrícia Froes Meyer1, Rodrigo Marcel Valentim da Silva2*,
Eneida de Morais Carreiro1, Fábio dos Santos Borges3,
Stephany Luanna Queiroga Farias4,
Thamires Santos Machado Figueiredo4 and Talita Duarte Martins4

1University Center of Rio Grande do Norte (UNI-RN), Natal, RN, Brazil.
2Estácio de Sá University, Natal, RN, Brazil.
3Estácio de Sá University, Rio de Janeiro, Brazil
4Potiguar University (UNP), Natal, RN, Brazil.
*Corresponding author's E-mail:
rodrigomarcelvalentim@gmail.com

Abstract

BACKGROUND: Cryolipolysis with plates is a method of applying cooling without a vacuum system, which can be used in regions with less chance of forming a "crease."  OBJECTIVE: To investigate the effects of cryolipolysis using a plate-shaped applicator (Crioplace™) in the treatment of fat.  MATERIALS AND METHODS: This is an experimental study in which women aged 25 to 45 years with adiposity located in the abdomen participated. Two applications of 75 min were made, using 04 plates in the abdomen regions, with -4°C being programmed as a temperature parameter. Anthropometric and ultrasound assessments were performed, and a satisfaction questionnaire on the validated treatment was conducted. The reassessments were performed 30 and 60 days after the first intervention. RESULTS: A reduction in adiposity was observed in the measurements of perimetry, plicometry and abdominal ultrasound (p<0.05). It was found that about 62.5% of the volunteers reported an improvement in water retention, about 62.5% reported the presence of loose clothing, and 31.3% reported satisfaction with the results obtained. It was observed that 18.5% of the volunteers reported that the treatment was excellent. CONCLUSION: The Crioplace™ method proved to be effective in reducing adiposity, with a high clinical satisfaction with the reduction in body measurements.

Keywords: adipose tissue; cryotherapy; ultrasound.

 

 

 

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CryoLetters 42 (5), 290-299 (2021)
© CryoLetters,
editor@cryoletters.org

AMMONIUM-FREE MEDIUM IS CRITICAL FOR REGENERATION OF SHOOT TIPS OF THE ENDANGERED SPECIES Pogostemon yatabeanus CRYOPRESERVED USING DROPLET-VITRIFICATION

Hyoeun Lee1, Hana Park1, Elena Popova2, Young-Yi Lee3, Sang-Un Park4 and Haeng-Hoon Kim1*

Department of Agricultural Life Science, Sunchon National University, Suncheon, 57922, Korea.
К.А. Timiryazev Institute of Plant Physiology of Russian Academy of Sciences, Moscow, 127276, Russia
National Agrobiodiversity Center, NIAS, RDA, Suwon, 16613, Korea
Division Plant Science and Resources, Chungnam National University, Daejeon, 34134, Korea.
* Corresponding author's E-mail:
cryohkim@scnu.ac.kr

Abstract

BACKGROUND: Pogostemon yatabeanus, synonym Dysophylla yatabeana, (Labiatae) is an endangered wild species in Korea. It has has a limited natural habitat and requires urgent conservation measures. OBJECTIVE: To develop an efficient cryopreservation protocol using in vitro shoot tips to complement traditional conservation approaches in case seeds are unavailable, or insufficient in number for conservation programs. MATERIALS AND METHODS: Node-cutting induced shoot tips of in vitro plants were produced and cryopreserved using a droplet-vitrification method following improvements in preculture, osmoprotection, vitrification solution (VS) and regrowth treatments. The starting protocol included preculture with 10% sucrose for 31 h, followed by osmoprotection with C4-35% (17.5% glycerol + 17.5% sucrose) for 40 min, and cryoprotection with A3-80% (33.3% glycerol + 13.3% DMSO + 13.3% EG + 20.1% sucrose) for 60 min on ice, cooling and warming using aluminum foil strips, and regrowth in MS hormone-free medium.  RESULTS: Shoot tips of Pogostemon yatabeanus were sensitive to the osmotic stress evidenced by low survival after step-wise preculture with 17.5% sucrose and cryopreservation without osmoprotection. Among VS tested, including PVS2, PVS3 and their alternatives, A3-80% on ice for 60 min resulted in the highest post-cryopreservation survival (80%) and regeneration (20%). Post-cryopreservation regeneration significantly improved (up to 73%) by incubation of cryopreserved shoot tips on ammonium-free medium followed by GA3–containing medium and medium without growth regulators. CONCLUSION: Cryopreservation of in vitro shoot tips using droplet-vitrification was developed as a complementary conservation approach for D. yatabeana. Adjustment of medium composition during the recovery stage was important for regeneration of healthy plants from both cryoprotected-control and cryopreserved shoot tips.

Keywords: alternative vitrification solutions; cryopreservation; endangered species conservation; osmoprotection; regrowth; systematic approach.

 

 

 

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CryoLetters 42 (5), 300-308 (2021)
© CryoLetters,
editor@cryoletters.org

CRYOPRESERVATION OF A THREATENED MEDICINAL PLANT, Valeriana jatamansi Jones, USING VITRIFICATION AND ASSESSMENT OF BIOSYNTHETIC STABILITY OF REGENERANTS

Shailika Sharma1, Kiran Parasher1, Papiya Mukherjee1*
and Yash Pal Sharma2

1Department of Botany, Panjab University, Chandigarh, 160 014, India.
2Department of Forest Products, Dr. YS Parmar University of Horticulture and Forestry, Solan, Himachal Pradesh, 173 230, India.
*Corresponding author's E-mail:
papiya23@gmail.com; pmukherjee@pu.ac.in

Abstract

BACKGROUND: Valeriana jatamansi Jones is a medicinal plant of the Himalayan region with high trade value. Since overexploitation of this wild species led it to be listed as threatened, a comprehensive conservation strategy is needed. Cryopreservation would be a useful complementary method to conventional conservation methods. OBJECTIVE: To develop a cryopreservation protocol for V. jatamansi with maintenance of biosynthetic stability of regenerants. MATERIALS AND METHODS: In vitro shoot tips were cryopreserved using vitrification with either PVS2 or PVS3 and the efficacy of the two cryoprotectant mixtures compared. Regenerated plantlets were evaluated by HPLC analysis for contents of four valepotriates viz. valtrate, acevaltrate, didrovaltrate and IVHD valtrate. RESULTS: The highest shoot recovery (91.6%) after transfer to liquid nitrogen was obtained when shoot tips were treated with PVS2 at 0°C for 110 min, which was significantly higher than the highest recovery (73.3%) obtained using PVS3 for any duration tested. Evaluation of biosynthetic stability showed no variation in valepotriate contents between in vitro maintained and cryopreserved derived plantlets. CONCLUSION: This protocol will be useful for the long-term conservation of this species as high frequency recovery and biosynthetic stability after cryopreservation were obtained.

Keywords: biosynthetic stability; HPLC; PVS; shoot tips; valepotriates; vitrification.

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