scholarly journals Relationship of extender and packaging system an the length of preservation and the quality of chilled semen of Boer goat

2016 ◽  
Vol 21 (1) ◽  
pp. 49
Author(s):  
Arie Febretrisiana ◽  
. Anwar ◽  
Simon Sinulingga

<p class="abstrak2">The aim of this research was to compare the effectiveness of different extender (either Triladyl or Tris Egg Yolk extender) and different packaging method (pool and straw) of chilled semen an the length of preservation and the quality of chilled semen of Boer goat. Semen was collected using an artificial vagina from 3 two years old Boer bucks with body weight of 50-55 kg. It was evaluated under a microscope, then each was diluted either in Tris egg yolk extender (TEY) or Triladyl. Those diluted sperms were then packed either in pool or straw and preserved at 5⁰C refrigerator. Sperm motility, viability and membrane integrity of each group were evaluated every 24 h for up to 5 days. Results showed that sperm motility in Triladyl of  pool packaging system up to 3 days was higher than straw packaging system or TEY in pool or straw packaging system which were 45.8%, 26.1%, 32.1% and 9.1%, respectively (P&lt;0.05). Percentage of sperm membrane integrity showed the same pattern to Triladyl both in pool and straw packaging system which was higher than TEY group (75.2% and 77,2%; P&lt;0.05). Sperm viability in Triladyl both in pool or straw packaging system decreased (P&lt;0.05) after 3 days of preservation (77.1% and 76.2%) but TEY significanly decreased after 4 days of preservation either in pool or straw packaging system (73.2% and 58.0%; P&lt;0.05). It was concluded that sperm quality decreased with increasing of the length of preservation while Triladyl extender in pool packaging system showed the best quality.</p><strong>Key Words: </strong>Chilled Semen, Boer, Triladyl, Tris Egg Yolk, Straw

2016 ◽  
Vol 28 (12) ◽  
pp. 1990 ◽  
Author(s):  
D. Acha ◽  
M. Hidalgo ◽  
I. Ortiz ◽  
M. J. Gálvez ◽  
J. J. Carrasco ◽  
...  

The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P < 0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P < 0.001), whereas DMSO affected sperm motility and membrane integrity (P < 0.001). DMFA 2.5% yielded higher (P < 0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.


2019 ◽  
Vol 6 (1) ◽  
pp. 1
Author(s):  
Muhammad Riyadhi ◽  
Anis Wahdi ◽  
Muhammad Rizal

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


2019 ◽  
Vol 6 (1) ◽  
pp. 78
Author(s):  
Muhammad Ade Salim ◽  
Muhammad Nur Ihsan ◽  
Nur Isnaini ◽  
Trinil Susilawati

ABSTRAKAir kelapa muda varietas viridisdapat dijadikan pengencer aletrnatif semen cair bagi program IB di daerah minim sarana semen beku. Tujuan penelitian ini untuk menguji pengaruh penggunaan air kelapa muda viridissebagai bahan pengencer terhadap kualitas semen cair kambing Boer setelah didinginkan. Dilaksanakanselama 3 bulan di Laboratorium Fakultas Peternakan UBUnit SumberSekar,Malang. Metodenya yaitu eksperimen. Semen dari  3 pejantan Boer umur 3-5 tahun, dikoleksi seminggu sekali dengan VB. Air kelapa mudaviridis umur 5-7 bulan serta tris aminomethane sebagai kontrol. Didesain menggunakan Rancangan Acak Kelompok (RAK) dengan 2 perlakuan yaitu P0 (tris aminomethane + 10% KT) dan  P1 (air kelapa muda viridis + 10% KT) masing-masing diulang 10 kali. Data dianalisis dengan analisis Ragam (Anova) dengan software Genstat 18. Variabelnya yaitu motilitas individu, viabilitas dan abnormalitas. Hasil penelitian yaitu motilitas individu pada P1bertahan sampai 4 hari (40,5± 24,3%), viabilitas terbaik sampai hari ke-5 (42±24,6%), abnormalitas terendah di hari ke-7(1,31± 0,6). Kesimpulannya, Pengencer air kelapa muda viridis dapat mempertahankan kualitas semen cair kambing Boer selama 4 hari untuk motilitas dan 5 hari untuk viabilitas.Kata Kunci:pengencer, air kelapa, varietas viridisABSTRACTYoung viridis coconut water could be used as an alternative to liquid semen diluent for artificial insemination program in the area with limited facility for frozen semen production. This study evaluated the use of young coconut water as a diluent on liquid semen quality of Boer goat after cold storage. This study was carried out for 3 months at Sumber Sekar Laboratory, Faculty of Animal Husbandry, University of Brawijaya, Malang. The semen was collected from 3 Boer bucks aged at 3 to 5 years old. The semen collection was done once a week with the aid of artificial vagina. The diluents used were young Viridis coconut (5 to 7 months old) and tris aminomethane. The method used was an experiment in a randomized block design with 2 treatments and 10 replicates. The treatments used were T0: tris aminomethane + 10% egg yolk (control) and T1:  young Viridis coconut water + 10% egg yolk. Data were analyzed by analysis of variance using Genstat 18 software. The variables measured were sperm individual motility, viability, and abnormality. The results showed that the sperm individual motility in T1 survived up to 4 days (40.5± 24.3%), the best viability at 5 days (42.0±24.6%),  while the lowest abnormality at 7 days (1.31±0.6). It could be concluded that: 1. Tris aminomethane diluent has higher quality with the storage length up to 9 days, 2. Young Viridis coconut water diluent could preserve liquid semen quality of Boer goat up to 4 days for sperm motility and 5 days for sperm viability.Keywords: diluents, coconut water, viridis variety


2020 ◽  
Vol 41 (1) ◽  
pp. 167 ◽  
Author(s):  
Breno Fernandes Barreto Sampaio ◽  
Bruno Gomes Nogueira ◽  
Maria Inês Lenz Souza ◽  
Eliane Vianna da Costa-e-Silva ◽  
Carmem Estefânia Serra Neto Zúccari

Plasma membrane composition has impact on phase transition from liquid crystal to gel state of cooled sperm cell. The incorporation of polyunsaturated fatty acids increases its fluidity and can contribute to sperm motility. The aim of this study was to compare the effect of adding docosahexaenoic acid (DHA) and ?-tocopherol (?-Toh) to the cooling extender, singly or combined, to the equine sperm parameters, submitted to cooling, up to 72 hours. Two ejaculates of ten stallions collected with artificial vagina were used, and evaluated for motility, plasma membrane integrity, chromatin fragmentation, mitochondrial activity and lipid peroxidation, according to the following treatments: C; DHA; ?-Toh; DHA/?-Toh; EtOH 100: and EtOH 140 (corresponding to control; 10 ng mL-1 of DHA; 2 mM of ?-Toh; : 10 ng mL-1 of DHA + 2 mM of ?-Toh; 100 µL of ethanol and 140 µL of ethanol respectively). DHA treatment showed higher motility (68.2 ± 12.3; p < 0.05) when compared to control (62.1 ± 16.2), DHA/?-Toh (61.3 ± 12.7) and EtOH (58.1 ± 8.6) groups. In lipid peroxidation assay, the control group showed 2,506.2 ± 796.4 ng of MDA 108 spermatozoa-1, being significantly higher (p < 0.05) than the groups treated with DHA (2,036.0 ± 687.0), ?-Toh (1,890.8 ± 749.5) and DHA/?-Toh (1,821.1 ± 627.2). In conclusion, ?-Toh was effective in diminishing lipid peroxidation of equine sperm subjected to cooling, and DHA improved sperm motility and, in spite of being a polyunsaturated fatty acid with high susceptibility to peroxidation, reduced lipid peroxidation.


Author(s):  
Trinil Susilawati ◽  
Feri Eka Wahyudi ◽  
Inna Anggraeni ◽  
Nurul Isnaini ◽  
Muhammad Nur Ihsan

This study aims to determine the effect of the substitution of bovine serum albumin (BSA) with cattle blood serum and egg white in the diluent of Cauda epididymal Plasma 2 (CEP-2) on sperm quality of Limousin cattle during cooling at 3-5 C. The research material used was rejected Limousin bull sperm (motility of 50-60%) from Artificial Insemination Centre Singosari, Malang. This research was a laboratory experiment using a randomized block design which was composed of six treatments with 10 replications, those were T0 as controls ((90% CEP-2 with BSA + 10% egg yolk); T1 (83.84% CEP-2 + 6.16% cattle blood serum + 10% egg yolk); T2 (81.84% CEP-2 + 8.16% cattle blood serum + 10% egg yolk); dan T3 (90% CEP-2 + 0,4% egg white + 10% egg yolk); T4 (90% CEP-2 + 0.8% egg white + 10% egg yolk); and T5 (90% CEP-2 without BSA + 10% egg yolk). Parameters measured were the percentage of motility, viability, and abnormality of sperms. Results of research after 48 hours of storage showed that the percentage of sperm motility in T0, T1, T2, T3, T4, and T5 were 40.50±5.90, 36±36.16, 34.00±6.58, 40.50±3.69, 38.50±3.37, and 38.50±4.12, respectively, while the percentage of sperms viability were 75.16±4.30, 70.50±2.88, 73.80±2.80, 74.80±3.30, 75.13±3.13, and 74.03±4.13, respectively, and the percentage of sperms abnormality were 10.14±2.34, 10.62±1.34, 11.33±2.00, 10.94±2.82, 10.02±1.95, and 10.78±1.96, respectively. In conclusion, CEP-2 diluent with or without the addition of 19% egg yolk in BSA and the substitution of BSA with 0.4-0.8% egg white can maintain semen quality to hour of 48 in cold storage.This study aims to determine the effect of the substitution of bovine serum albumin (BSA) with cattle blood serum and egg white in the diluent of Cauda epididymal Plasma 2 (CEP-2) on sperm quality of Limousin cattle during cooling at 3-5 C. The research material used was rejected Limousin bull sperm (motility of 50-60%) from Artificial Insemination Centre Singosari, Malang. This research was a laboratory experiment using a randomized block design which was composed of six treatments with 10 replications, those were T0 as controls ((90% CEP-2 with BSA + 10% egg yolk); T1 (83.84% CEP-2 + 6.16% cattle blood serum + 10% egg yolk); T2 (81.84% CEP-2 + 8.16% cattle blood serum + 10% egg yolk); dan T3 (90% CEP-2 + 0,4% egg white + 10% egg yolk); T4 (90% CEP-2 + 0.8% egg white + 10% egg yolk); and T5 (90% CEP-2 without BSA + 10% egg yolk). Parameters measured were the percentage of motility, viability, and abnormality of sperms. Results of research after 48 hours of storage showed that the percentage of sperm motility in T0, T1, T2, T3, T4, and T5 were 40.50±5.90, 36±36.16, 34.00±6.58, 40.50±3.69, 38.50±3.37, and 38.50±4.12, respectively, while the percentage of sperms viability were 75.16±4.30, 70.50±2.88, 73.80±2.80, 74.80±3.30, 75.13±3.13, and 74.03±4.13, respectively, and the percentage of sperms abnormality were 10.14±2.34, 10.62±1.34, 11.33±2.00, 10.94±2.82, 10.02±1.95, and 10.78±1.96, respectively. In conclusion, CEP-2 diluent with or without the addition of 19% egg yolk in BSA and the substitution of BSA with 0.4-0.8% egg white can maintain semen quality to hour of 48 in cold storage.


Animals ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 999 ◽  
Author(s):  
Ayman Abdel-Aziz Swelum ◽  
Islam M. Saadeldin ◽  
Hani Ba-Awadh ◽  
Mohsen G. Al-Mutary ◽  
Abdullah F. Moumen ◽  
...  

This study compared the efficiency of commercial egg yolk-free (AndroMed, OPTIXcell) and egg yolk-supplemented (Triladyl, Steridyl) Tris-based extenders for semen cryopreservation in seven adult dromedary camels. The camel-specific extender SHOTOR was used as control. The collected semen samples were evaluated and diluted with SHOTOR, Triladyl, Steridyl, AndroMed, or OPTIXcell. The diluted semen was gradually cooled and equilibrated for two hours before liquid nitrogen freezing. Semen was evaluated prior to freezing and after freeze-thawing cycles for motility, kinetics, vitality, abnormality, plasma membrane integrity, and DNA fragmentation using computer-assisted sperm analysis. In pre-freezing evaluation, progressive sperm motility was higher in SHOTOR-diluted samples (21.54 ± 1.83) than in samples diluted with Steridyl, OPTIXcell, or AndroMed (15.76 ± 1.80, 17.43 ± 1.10, and 13.27 ± 1.07, respectively). Moreover, Triladyl and SHOTOR resulted in significantly (p < 0.05) better sperm vitality and DNA integrity than all other diluents, but Triladyl resulted in a significantly (p < 0.05) better plasma membrane integrity (87.77 ± 0.31) than SHOTOR (85.48 ± 0.58). In the post-thawing evaluation, Triladyl led to significantly (p < 0.05) higher sperm motility (38.63 ± 0.81%; p < 0.05) when compared to SHOTOR, Steridyl or AndroMed (35.09 ± 1.341%, 34.4 ± 0.84%, and 31.99 ± 1.48%, respectively), with OPTIXcell being the least efficient (28.39 ± 0.86%). Progressive sperm motility was the highest when using Triladyl. Post-thawing curvilinear, straight line and average path sperm velocities were highest with Triladyl and lowest with AndroMed. Triladyl led to the highest linearity coefficient and straightness sperm coefficient, while SHOTOR to the highest DNA and plasma membrane integrity. OPTIXcell and AndroMed resulted in poor post-thawing sperm vitality, while Steridyl was less efficient than Triladyl. The highest rate of sperm abnormalities was recorded with OPTIXcell and the lowest with SHOTOR or Triladyl. In conclusion, SHOTOR, Triladyl, Steridyl, AndroMed, and OPTIXcell can all be used for camel semen cryopreservation; however, SHOTOR and Triladyl provided the best post-thawing sperm quality. Based on our findings, Triladyl is the best commercially available extender for dromedary camel semen cryopreservation to date.


2021 ◽  
Vol 8 ◽  
Author(s):  
Jiahao Zou ◽  
Lixuan Wei ◽  
Dexian Li ◽  
Yongtao Zhang ◽  
Guang Wang ◽  
...  

In the process of cryopreservation of dairy goat semen, it will face many threats such as oxidative damage, which will affect the motility and plasma membrane function of sperm. As an endogenous antioxidant in animals, glutathione (GSH) can significantly improve the quality of thawed sperm when added to the frozen diluent of semen of pigs and cattle. In this study, different concentration gradients of GSH [0 mmol/L (control), 1, 2, 3, 4 mmol/L] were added to the frozen diluent of Guanzhong dairy goat semen. By detecting the sperm motility parameters, acrosome intact rate and plasma membrane intact rate after thawing, the effect of GSH on the cryopreservation of dairy goat semen was explored. Sperm motility parameters were measured with the computer-aided sperm analysis (CASA) system (total power, TM; forward power, PM; linearity, LIN; average path speed, VAP; straight line speed, VSL; curve speed, VCL; beat cross frequency, BCF). The sperm acrosome integrity rate after thawing was detected by a specific fluorescent probe (isothiocyanate-labeled peanut agglutinin, FITC-PNA), and the sperm plasma membrane integrity rate after thawing was detected by the hypotonic sperm swelling (HOST) method. Reactive oxygen species (ROS) kit, malondialdehyde (MDA) kit, superoxide dismutase (SOD) kit, glutathione peroxidase (GSH-PX) kit were used to detect various antioxidant indicators of thawed sperm. in vitro fertilization experiment was used to verify the effect of adding glutathione on sperm fertilization and embryo development. The results showed that when the concentration of glutathione was 2 mmol/l, the sperm viability, plasma membrane intact rate, and acrosome intact rate were the highest after thawing, reaching 62.14, 37.62, and 70.87% respectively, and they were all significantly higher. In terms of antioxidant indexes; the values of SOD and GSH-PX were 212.60 U/ml and 125.04 U/L, respectively, which were significantly higher than those of the control group; The values of ROS and MDA were 363.05 U/ml and 7.02 nmol/L, respectively, which were significantly lower than the control group. The addition of 2 mmol/L glutathione significantly improves the fertilization ability of sperm. In short, adding 2 mmol/l glutathione to the semen diluent can improve the quality of frozen Guanzhong dairy goat sperm.


2012 ◽  
Vol 12 (2) ◽  
pp. 7-13
Author(s):  
Dasrul Dasrul ◽  
Rasmaidar Rasmaidar ◽  
Abdul Harris

Effect of vitamin E addition (alfa-tokoferol) into sperm washing medium by centrifuge on the quality of Brahman cattle spermatozoa ABSTRACT. The aims of study to determine the effectiveness of the addition of vitamin E in the washing medium by centrifugation on sperm quality Brahman cattle. frozen semen of Brahman cattle, divided into 4 treatment groups addition of vitamin E in the washing medium: 0.0gr/100 ml medium (K0), 0.1gr /100 ml medium (K1); 0.2gr/100 ml medium (K2) and 0.3 g / 100 ml medium (K4), each group was repeated 5 times. Examination of motility, viability and integrity of sperm membrane performed according to WHO standards. The data obtained were analyzed with one-way ANOVA and Duncan test. The average percentage of motility, viability and membrane integrity of spermatozoa in the addition of vitamin E were significantly different (P 0.05) compared to the control. Percentage of motility, viability and membrane integrity of spermatozoa in the group K2 significantly higher (P 0.05) compared with the group K3: K1 and K0. Percentage of motility, viability and sperm capacitation and sperm live on group K3 significantly higher (P 0.05) compared with the K1 and K0. While the percentage motility of spermatozoa in the group K1 higher were not significant (P 0.05) compared with the group K0. The addition of vitamin E in the medium on the process of washing spermatozoa Brahman cattle. The addition of vitamin E 0.2gr/100ml better than vitamin E 0.1gr/100ml and 0.3gr/100ml in maintaining the percentage of motility and live spermatozoa Brahman cattle.


2018 ◽  
Vol 63 (1) ◽  
pp. 53-65
Author(s):  
Ezekiel Adekunle ◽  
James Daramola ◽  
Olusiji Sowande ◽  
John Abiona ◽  
Monsuru Abioja

This study investigated the effects of apple and orange juices on quality of refrigerated spermatozoa of goat bucks. Semen samples from WAD goat bucks were diluted with Tris-egg yolk extenders each supplemented with apple and orange juices at 0, 2.5, 5, 7.5 and 10/100 ml of diluents. The diluted semen samples were assessed for sperm viability and malondialdehyde (MDA) concentration after in vitro storage for 240 hours at 5oC. The ability to maintain sperm motility was higher in the extenders with 7.5% orange juice followed by 10% apple juice compared to other treatments (P<0.05). The extenders supplemented with 2.5%, 5% and 7.5% apple juice, and 5% orange juice had higher intact acrosome compared to other treatments and the control (P<0.05). The 10% orange juice had higher percentage membrane integrity compared to other treatments. Consistent and reduced (P<0.05) MDA levels were observed in the extenders supplemented with fruit juices and lower MDA was observed in the extenders supplemented with 10% apple juice compared to other treatments and the control (P<0.05). The findings reveal that additions of the fruit juices to semen extenders to maintain the viability of refrigerated spermatozoa were best at concentrations of 10 ml/100 ml of apple juice and 7.5 ml/100 ml of orange juice.


2016 ◽  
Vol 28 (6) ◽  
pp. 663 ◽  
Author(s):  
Noelia Prieto-Martínez ◽  
Ingrid Vilagran ◽  
Roser Morató ◽  
Joan E. Rodríguez-Gil ◽  
Marc Yeste ◽  
...  

Aquaporins (AQPs) are integral membrane water channels that allow transport of water and small solutes across cell membranes. Although water permeability is known to play a critical role in mammalian cells, including spermatozoa, little is known about their localisation in boar spermatozoa. Two aquaporins, AQP7 and AQP11, in boar spermatozoa were identified by western blotting and localised through immunocytochemistry analyses. Western blot results showed that boar spermatozoa expressed AQP7 (25 kDa) and AQP11 (50 kDa). Immunocytochemistry analyses demonstrated that AQP7 was localised in the connecting piece of boar spermatozoa, while AQP11 was found in the head and mid-piece and diffuse labelling was also seen along the tail. Despite differences in AQP7 and AQP11 content between boar ejaculates, these differences were not found to be correlated with sperm quality in the case of AQP7. Conversely, AQP11 content showed a significant correlation (P < 0.05) with sperm membrane integrity and fluidity and sperm motility. In conclusion, boar spermatozoa express AQP7 and AQP11, and the amounts of AQP11 but not those of AQP7 are correlated with sperm motility and membrane integrity.


Sign in / Sign up

Export Citation Format

Share Document