Efficacy of egg yolk based and egg yolk free soya bean milk based extenders for cryopreservation of Zebu cattle and buffalo semen

2018 ◽  
Author(s):  
P. J. Chaudhary ◽  
A. J. Dhami ◽  
D. V. Chaudhari ◽  
K. K. Hadiya ◽  
J. A. Patel
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Zebu Cattle ◽  
Motile Sperm ◽  
Comparative Efficacy ◽  
Buffalo Semen ◽  
Sample Technique ◽  
Live Sperm ◽  
Surti Buffalo

This study was undertaken on three mature bulls each of Gir cattle and Surti buffalo breeds to evaluate the comparative efficacy of egg yolk based standard TFYG (Tris-citrate-fructose-yolk-glycerol) extender and egg yolk free soybean based commercial extenders Optixcell® (IMV, France) and Andromed® (Minitube, Germany) under split-sample technique. The ejaculates (9/bull) were extended @ 100×106 sperm ml-1 with three extenders and frozen using biofreezer following 4 hr of equilibration. The pooled means of progressively motile sperm observed (irrespective of extenders) at initial, pre-freeze and post-thaw stage in Gir bulls semen were 76.53±0.53, 71.11±0.53 and 39.86±0.90% and in Surti buffalo 80.76±0.39, 74.65±0.45 and 40.35±1.07%, respectively. The corresponding values for live sperm were 75.64±0.76, 69.01±0.97 and 47.99±1.11 % for Gir and 80.90±0.45, 75.76±0.48 and 52.33±0.86 % for Surti buffalo; and those of intact acrosome 94.29±0.25, 90.29±0.27 and 79.29±0.33 % for Gir bulls, and 93.94±0.21, 89.94±0.23 and 78.95±0.26 % for Surti buffalo semen, respectively. The HOS reactive sperm at initial, pre-freeze and post-thaw stage were 76.18±0.74, 71.04±0.76 and 27.90±0.70 % for Gir, and 81.83±0.35, 76.47±0.39 and 27.83±0.68 % for Surti bulls, respectively. The overall mean post-thaw incubation (37°C) survival of spermatozoa observed at 60, 120 and 180 min were 28.40±0.91, 17.78±0.86 and 9.44±0.72% for Gir bulls semen, and 28.01±0.99, 18.40±1.01 and 10.51±0.93% for Surti buffalo semen, respectively. Optixcell was proved superior, and at par with TFYG, than the Andromed in maintaining greater motility, viability, morphology, acrosomal/plasma membrane integrity including post-thaw sperm longevity of cattle and buffalo spermatozoa with significant differences only in sperm motility and post-thaw longevity. The motile, live and HOST reactive sperm were significantly higher in buffalo semen than cattle at initial and pre-freeze stage, but not at post-thaw stage. The results showed that egg yolk free commercial Optixcell extender and egg yolk based TFYG extender were at par in terms of most of the sperm quality traits, hence any one of them can be preferred over Andromed for successful routine cryopreservation of cattle and buffalo semen.

Evaluation of comparative efficacy of egg yolk and soya based extendersfor refrigeration preservation (5ºC) of buffalo semen

2016 ◽  
Author(s):  
A.J. Dhami ◽  
D.V. Chaudhari ◽  
J. A. Patel ◽  
K. K. Hadiya
Keyword(s):  
Sperm Quality ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Storage Duration ◽  
Normal Sperm ◽  
Buffalo Semen ◽  
Abnormal Head ◽  
Live Sperm ◽  
Surti Buffalo

The aim of this study was to compare commercially available soybean based extenders, viz. Bioxcell® and Optixcell® (IMV, France) with standard Tris-citrate-fructose-egg yolk-glycerol (TFYG) extender for refrigeration preservation of buffalo semen. Forty eight good quality ejaculates (8/bull) having initial motility >70% obtained at weekly interval in AV from 6 mature Surti buffalo bulls were split-diluted (at 34°C keeping 100×106 sperm ml-1) in TFYG, Bioxcell and Optixcell extenders and stored in refrigerator at 5°C for 72 hrs. The mean percentages of progressively motile spermatozoa observed on dilution (0-hr), and after 24, 48, 72 hrs of refrigeration preservation of semen at 5°C were 78.54±0.30, 68.30±0.30, 59.38±0.35 and 51.63±0.47; live sperm 90.48±0.19, 76.89± 0.51, 69.51±0.35 and 61.86±0.45; morphologically normal sperm 93.85±0.08, 90.70±0.11, 89.08±0.10 and 87.66±0.10; intact acrosome 94.40± 0.12, 88.22±0.13, 84.60±0.14 and 81.70±0.17, and HOS reactive sperm 79.35 ±0.24, 65.99±0.31, 57.69±0.36 and 50.70±0.43, respectively, all of which decreased gradually and highly significantly (P is less than 0.01) with increase in storage time. The results were significantly (Pis less than 0.05) superior with Optixcell followed by TFYG extender than with the Bioxcell. The overall mean percentages of sperms with abnormal head, mid-piece and tail and those with different forms of acrosomal defects like swollen, ruffled, detached and denuded acrosome also followed the pattern of morphologically normal sperm and intact acrosomes. The values of all these traits increased significantly (Pis less than 0.05) with increase in storage duration at 5°C. The sperm protective ability of Optixcell extender was superior followed by TFYG and the least of Bioxcell. The Optixcell, a recently launched improved soybean milk based extender, excelled the Bioxcell extender and was statistically similar to TFYG in respect of all the sperm quality parameters to satisfactorily preserve the quality of buffalo semen at refrigeration temperature up to 3 days.

Quality of Labrador- Retriever dog semen on short-term preservation in different extenders

2017 ◽  
Author(s):  
Arunoday Das ◽  
R. K. Biswas ◽  
B. C. Deka ◽  
D. J. Dutta
Keyword(s):  
Citric Acid ◽  
Egg Yolk ◽  
Sperm Head ◽  
Labrador Retriever ◽  
Comparative Efficacy ◽  
Short Term ◽  
Sample Technique ◽  
Digital Manipulation ◽  
Live Sperm

The objective of the present study was to find the comparative efficacy of three extenders to preserve semen of Labrador-Retriever (LR) dogs at 5o C for a short term. The semen samples of LR dogs were collected by digital manipulation method and extended at the rate of 1:4 in Tris-Egg Yolk- Citric Acid-Glucose (TEYCAG), Tris-Egg Yolk- Citric Acid-Fructose (TEYCAF) and Egg Yolk-Citrate-Glycine-Glucose (EYCGG) extenders by split sample technique. Semen was evaluated at 0, 24, 48, 72, 96 and 120 hours of preservation. Mean motile, live and HOST-reacted sperm and acrosomal, head, mid piece and tail abnormalities of spermatozoa varied significantly (PP less than 0.01) between extenders and between preservation periods. The interactions between extender and preservation period were also significant (P less than 0.01) except for HOST-reacted and head abnormalities of sperm. The highest mean motile, live and HOST-reacted sperm were recorded in TEYCAG extender which did not differ significantly from that of TEYCAF extender. Mean per cent sperm acrosomal and tail abnormalities were significantly (PP less than 0.05) lower, and the incidences of mean sperm head and mid piece abnormalities were also lower in TEYCAG, but not significantly from that in TEYCAF irrespective of hour of preservation. Per cent motile, live and HOST-reacted sperm were significantly (PP less than 0.05) lower and sperm acrosomal, head, mid piece and tail abnormalities were significantly (PP less than 0.05) higher in EYCGG as compared to that in TEYCAG and TEYCAF irrespective of hour of preservation. It was concluded that the semen of LR dog sustained good quality during preservation up to 5 days at 5oC suitable for successful artificial insemination and would be preserved better in TEYCAG and TEYCAF extenders than in EYCGG extender, since more than 50 per cent sperm motility and live sperm were maintained up to 120 hours of preservation in the former two extenders.

scholarly journals Comparative Study of Gir Cattle and Surti Buffalo Bulls Semen under Middle Gujarat Climate

2017 ◽  
Vol 13 (01) ◽  
Author(s):  
P. J. Chaudhary ◽  
A. J. Dhami ◽  
D. V. Chaudhari ◽  
K. K. Hadiya ◽  
J. A. Patel
Keyword(s):  
Sperm Concentration ◽  
Membrane Integrity ◽  
Mean Values ◽  
Abnormal Sperm ◽  
Buffalo Semen ◽  
The Mean ◽  
Mass Activity ◽  
Live Sperm ◽  
Surti Buffalo ◽  
Fresh Semen

This study was undertaken during the favourable breeding season of the year 2016-17 on healthy mature Gir cattle and Surti buffalo bulls, three each, at Sperm Station of the College. The ejaculates (9/bull, total 54) collected in the morning using artificial vagina were evaluated for routine seminal attributes, including acrosomal and plasma membrane integrity. The mean values of ejaculate volume, sperm concentration, mass activity (0-5 score), individual sperm motility, live sperm, abnormal sperm, intact acrosome and HOST reactive sperms observed in fresh semen of Gir cattle and Surti buffalo bulls were 6.20±0.42 and 3.34±0.23 ml (P less than 0.01), 1169.44±61.71 and 846.30±54.82 million (P less than 0.01), 3.44±0.09 and 3.42±0.08, 76.53±0.53 and 80.76±0.39 % (P less than 0.05), 81.00±1.32 and 84.73±0.78 % (P less than 0.05), 6.00±0.37 and 5.81±0.40 %, 95.59±0.35 and 95.54±0.25 % as well as 80.30±1.90 and 84.58±0.88 % (P less than 0.05), respectively. The variation between cattle and buffalo semen was significant for most of these traits. The variations between bulls within breed were however not significant. Significant correlations were observed only between mass activity and initial motility (0.62), and live and abnormal sperm (-0.41) in Gir bulls, and for ejaculate volume with sperm concentration (-0.56) and abnormal sperm (0.45), and between live and HOS reactive sperms (0.48) in Surti bulls.

Kriopreservasi Semen Kambing Boer dengan Konsentrasi Pengencer Nira Aren dan Gliserol Berbeda

2019 ◽  
Vol 6 (1) ◽  
pp. 1
Author(s):  
Muhammad Riyadhi ◽  
Anis Wahdi ◽  
Muhammad Rizal
Keyword(s):  
Sperm Motility ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Boer Goat ◽  
Palm Juice ◽  
Sperm Membrane ◽  
Live Sperm

ABSTRAK                                                                        Penelitian bertujuan untuk mengetahui efektivitas nira aren sebagai pengencer alternatif dalam proses pembekuan (kriopreservasi) semen kambing boer.Kriopreservasi semen kambing boer menggunakan pengencer tris-gliserol-kuning telur (P1 73-7-20%), nira aren-gliseol-kuning telur(masing-masing P2 74-6-20%, P3 73-7-20%, dan P4 72-8-20%) dan andromed (P5 tanpa mengandung kuning telur dan gliserol). Parameter evaluasi meliputi motilitas, viabilitas, dan membrane plasma utuh setelah pengenceran, ekuilibrasi dan thawing.  Evaluasi motilitas pasca thawing menunjukkan P5 52% berbeda nyata (P<0.05) dengan P1 42%, selanjutnya P5 dan P1 berbeda sangat nyata (P<0.05) dengan P2 8%, P3 6% dan P4 12%.  Viabilitas pasca thawing menunjukkan P5 65,4% tidak berbeda nyata (P>0,05) dengan P1 61,8%, akan tetapi P5 dan P1 berbeda sangat nyata (P<0.05) dengan P2 26,2%, P3 29,8%, dan P4 34%.  Membran plasma utuh (MPU) pasca thawing menunjukkan P5 66,2% tidak berbeda nyata (P>0,05) dengan P1 65,4%, akan tetapi keduanya berbeda sangat nyata (P<0.05) dengan P2 39%, P3 38%, dan P4 36,2%.  Disimpulkan kriopreservasi semen kambing boer dengan pengencer nira aren dan gliserol pada konsentrasi berbeda belum dapat dipergunakan sebagai sumber bibit berdasarkan standar nasional Indonesia.Kata Kunci : Kambing boer, semen, nira arenABSTRACTThe experiment was conducted to determine the effect of sugar palm juice as alternative extender for cryopreservation process of boer semen.Tris-glycerol-egg yolk (P1 73-7-20%), Sugar palm juice-glyserol-egg yolk (P2 74-6-20%, P373-7-20%, dan P4 72-8-20%), and andromed (P5) used as a extender  in the cryopreservation process of boer semen.  Sperm motility (%), live sperm (%) and sperm membrane integrity (%) were recorded after diluted, equilibration and freeze-thawing.  Result of post thawing motility showed that P5 52% was significantly different (P <0.05) with P1 42%, then P5 and P1 were significantly different (P <0.05) with P2 8%, P3 6% and P4 12%. Viability after thawing showed P5 65.4% was not significantly different (P> 0.05) with P1 61.8%, but P5 and P1 significantly different (P <0.05) with P2 26.2%, P3 29.8 %, and P4 34%. Spermmembrane integrity post-thawing showed P5 66.2% was not significantly different (P> 0.05) with P1 65.4%, but both were very significantly different (P <0.05) with P2 39%, P3 38% and P4 36.2%. Conclusions, sugar palm juice-glycerol-egg yolk with differentconcentrationsineffectively as an alternative extenderin cryopreservation of boer semen.Keywords: boer goat, semen, sugar palm juice

scholarly journals Effect of Supplementation of Polyvinylpyrrolidone in Extender on Buffalo Semen Parameters

2020 ◽  
Vol 53 (1) ◽  
Author(s):  
Bushra Ismail Khan ◽  
Shamim Akhter ◽  
Sanwal Aslam ◽  
Rabea Ejaz
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Bubalus Bubalis ◽  
Semen Parameters ◽  
Suction Pump ◽  
Bull Semen ◽  
Buffalo Semen ◽  
Live Sperm ◽  
And Control

The current study was planned to evaluate the supplementation of Polyvinylpyrrolidone in extender on cryopreservation of Nili-Ravi buffalo bull semen. The semen samples were collected from Nili-Ravi buffalo (Bubalus bubalis) bull kept at SPU Qadirabad, District Sahiwal, Pakistan. Qualifying semen ejaculates having motility >60%, volume >5-6ml and concentration >0.5 billion/ ml were diluted 50 × 106 motile sperm ml approximately at 37°C in Tris-citric acid extender supplemented with different concentrations of PVP (0.01, 0.05, 0.1mM). The extender without PVP was kept as control. Semen was stored at 4°C for a period of 2 h and kept at 4°C for 4h. Semen was filled in 0.5 ml French straws using suction pump at 4°C, plunged and stored in liquid nitrogen (-196°C). Semen straws were rewarmed at 37°C for 30 seconds and assessed for sperm motility, plasma membrane integrity (PMI), dead sperm percentage and the live sperm percentage. The data on the role of PVP on different parameters of semen quality were analyzed by using ANOVA and RCBD. Higher percentage (P< 0.05) of sperm motility (66.1±7.51 and 59.4±10.72) and PMI (72.9±5.39 and 75.7±6.5) was observed in extenders having 0.05 mM and 0.1mM PVP compared to extenders having 1.5mM PVP and control. The percentage acrosomal integrity was observed greater (P< 0.05) in extended semen containing 0.1mM (68.2±0.50) PVP compared to extenders having 0.01 and control.

Cryopreservation of collared peccary (Pecari tajacu L., 1758) epididymal sperm using extenders based on Tris and powdered coconut water (ACP®-116c)

Zygote ◽  
2018 ◽  
Vol 26 (4) ◽  
pp. 301-307 ◽  
Author(s):  
José A. B. Bezerra ◽  
Andréia M. Silva ◽  
Patrícia C Sousa ◽  
Lívia B. Campos ◽  
Érica C. G. Praxedes ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Coconut Water ◽  
Computer Assisted ◽  
Epididymal Sperm ◽  
Sperm Plasma Membrane ◽  
Pecari Tajacu ◽  
Functional Membrane

SummaryThe aim of this study was to establish a functional freezing–thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed. In addition, to evaluate the survival of frozen–thawed sperm, a thermal resistance test (TRT) was executed. Samples preserved using Tris were in better condition compared with those preserved using ACP®, showing higher values for most assessments performed, including CASA and the TRT (P<0.05). After determining Tris to be the better of the two extenders, additional samples were thawed using different thawing rates (37°C/60 s, 55°C/7 s, 70°C/8 s). Sperm thawed at 37°C/60 s had the greatest preservation (P<0.05) of viability (54.1 ± 5.9%) and functional membrane integrity (43.2 ± 5.4%), and had higher values for various CASA parameters. In conclusion, we suggest the use of a Tris-based extender added to egg yolk and glycerol for the cryopreservation of epididymal sperm obtained from collared peccaries. In order to achieve better post-thawing sperm quality, we suggest that samples should be thawed at 37°C/60 s.

scholarly journals Effect of N-Methylacetamide Concentration and Thawing Rate on Chicken Sperm Quality after Cryopreservation

Animals ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 824
Author(s):  
Fabio Mosca ◽  
Luisa Zaniboni ◽  
Ahmad Abdel Sayed ◽  
Nicolaia Iaffaldano ◽  
Dominga Soglia ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Kinetic Parameters ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Motile Sperm ◽  
Cell Membrane Integrity ◽  
Before And After ◽  
Freezing Procedure ◽  
Thawing Rate

In seeking alternative cryoprotectants to glycerol for a reference chicken semen freezing procedure, the aim of the present study was to compare the effect of two concentrations of N-Methylacetamide (MA) and two thawing rates on the quality of frozen-thawed semen. Semen samples were diluted in Lake pre-freezing extender, including 0.1 M trehalose in presence of 6% or 9% MA, loaded into straws, frozen in nitrogen vapors, and stored in liquid nitrogen. The following thawing treatments were used: 5 °C for 100 s and 38 °C for 30 s. Sperm quality (cell membrane integrity, motility and kinetic parameters) was assessed before and after cryopreservation. The decrease of MA concentration from 9 to 6% improved sperm quality after freezing/thawing and this effect was dependent on thawing temperature. Decreasing the MA concentration from 9 to 6% improved the proportion of undamaged membrane, motile, and progressive motile sperm recovered after thawing at 5 °C for 100 s; in contrast, no effect of the MA concentration was observed thawing at 38 °C for 30 s. Therefore, the treatment with 6% MA and thawing at 5 °C for 100 s has given the best cryoprotective action. These results contribute to improve the efficacy of the current chicken semen cryopreservation procedures.

scholarly journals Effect of the addition of sodium caseinate on the viability of cryopreserved buffalo semen

2020 ◽  
pp. 2209-2218
Author(s):  
Fernando Evaristo da Silva ◽  
Jaqueline Candido Carvalho ◽  
Camila de Paula Freitas Dell'Aqua ◽  
Frederico Ozanam Papa ◽  
Marc Roger Jean Marie Henry ◽  
...  
Keyword(s):  
Cell Damage ◽  
Membrane Integrity ◽  
Sodium Caseinate ◽  
Egg Yolk ◽  
Freezing Process ◽  
Computer Assisted ◽  
Semen Cryopreservation ◽  
Sperm Analysis ◽  
Frozen Semen ◽  
Buffalo Semen

The use of cooled semen in artificial insemination operations results in higher pregnancy rates than the use of frozen semen. This result seems to be related to the more severe damage triggered by the freezing process than that observed during refrigeration. Due to its ability to bind to sperm-binding proteins and calcium ions, sodium caseinate has been studied as a substance capable of preventing early sperm capacitation, a significant cause of the decreased pregnancy rate resulting from the use of frozen semen. The first objective of this study was to evaluate whether a commercial egg yolk diluent developed for frozen bovine semen could be used for buffalo semen cryopreservation; the second objective was to investigate the effect of this diluent in combination with sodium caseinate during the procedures of buffalo sperm cryopreservation using flow cytometry and computer-assisted sperm analysis. In the first part of the study, comparing the results of spermatic kinetics and plasma and acrosomal membrane integrity, it was observed that the freezing process resulted in more cell damage than the cooling process. In the second part of the study, no effects of the addition of sodium caseinate to the egg yolk diluent were observed. From the results of the present study, it was possible to conclude that the egg yolk-based diluent was suitable for buffalo semen cryopreservation and that the addition of sodium caseinate did not decrease the harmful effects related to seminal cryopreservation.

In vitro Supplementation of Linoleic Acid Improves Quality of Cryopreserved Buffalo Semen

2020 ◽  
Author(s):  
R Ejaz ◽  
S Qadeer ◽  
M S Ansari ◽  
B A Rakha ◽  
S Shamas ◽  
...  
Keyword(s):  
Linoleic Acid ◽  
Membrane Integrity ◽  
Sperm Chromatin ◽  
Standard Procedures ◽  
Buffalo Semen ◽  
Chromatin Integrity ◽  
Live Sperm ◽  
And Control

Present study was designed to evaluate the effect of linoleic acid (LA) supplementation in extender on post thaw quality of cryopreserved buffalo semen. Semen was collected from three adult Nili Ravi buffalo bulls of same age with artificial vagina (42°C) for five weeks (replicates; N=30). Qualified semen ejaculates (>1mL volume, >60% motility, >0.5 billion/mL concentration) were diluted in tris-citric acid extender containing 0.0 (control), 5.0, 10.0 and 20.0ng mL-1 of LA and were cryopreserved using standard procedures. Sperm motility and plasma membrane integrity were improved plessthan0.05) in extender containing 10.0 ng mL-1 of LA compared to other treatments and control while number of acrosome intact live sperm, chromatin integrity and number of morphologically normal sperms remained the same. In conclusion, LA supplementation in extender at 10.0 ng mL-1 was found to be beneficial to improve post thaw quality of cryopreserved buffalo semen.

scholarly journals Development of Sperm Cryopreservation Protocols for Sharks and Rays: New Tools for Elasmobranch Conservation

2021 ◽  
Vol 8 ◽  
Author(s):  
Pablo García-Salinas ◽  
Victor Gallego ◽  
Juan F. Asturiano
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Ex Situ ◽  
Conservation Strategies ◽  
Sperm Cryopreservation ◽  
In Captivity ◽  
Reproductive Techniques ◽  
Elasmobranch Species ◽  
Reproductive Cycles

Elasmobranchs are one of the most endangered vertebrate groups on the planet, but despite this situation the use of reproductive techniques in elasmobranch conservation strategies has been scarce. Among these techniques, sperm preservation is a potential tool for ex situ conservation and aquaria sustainability. However, there are no widespread preservation protocols for elasmobranch sperm, and shark sperm cryopreservation has never been achieved before. Here we present the establishment of successful cryopreservation protocols for elasmobranch sperm, tested in several species. We have formulated a sperm extender that can be used for different elasmobranch species, capable of maintaining sperm motility for several weeks. Additionally, we achieved the cryopreservation of sperm by previously diluting it in our extender and supplementing it with different combinations of cryoprotectants. The effects of methanol and dimethyl sulfoxide as permeating cryoprotectants were evaluated, as well egg yolk as a non-permeating cryoprotectant. Sperm quality was assessed by studying the motility and membrane integrity post-thawing, demonstrating its effectiveness in the 10 species tested, including two which are considered Critically Endangered. This is the first time that shark sperm cryopreservation has been reported, broadening our knowledge of the reproductive techniques that can be applied to elasmobranchs and laying the foundations for the first cryobanks for shark and ray sperm. Outcomes from this study will be useful for ex situ conservation efforts developed by public aquaria. A regular supply of frozen sperm will reduce the problems that result from the transport of specimens, inbreeding or lack of synchronized reproductive cycles in captivity.

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