scholarly journals THE ROLE OF GUAVA FRUIT FILTRATE ON MAINTAINING THE PRE-FREEZING AND POST-THAWED QUALITY OF BALI BULL SPERMATOZOA

Author(s):  
I Wayan Lanus Sumadiasa ◽  
Lukman Lukman ◽  
Rodiah Rodiah

This study aimed to determine the role of guava fruit filtrate (GFF) on maintaining the pre-freezing and post-thawed quality of Bali bullspermatozoa in tris-egg yolk (TEY) extender. The bovine semen was collected using artificial vagina twice a week (n= 10). Ejaculate sampleswere divided into four tubes and each tube was respectively added the following diluents: TEY, TEY + 5% GFF, TEY + 10% GFF, and TEY +15% GFF. The diluted semen was filled into the straws (0.25 mL) and cooled in refrigerator at 4° C for 2.5 hours. The samples were kept onvapor phase of liquid nitrogen (N2) (-120° C) for 10 minutes prior to be stored in liquid phase (-196° C). The quality of pre-freezing and postthawedspermatozoa in straw sample (i.e. motility, viability, and abnormality) was evaluated under light microscopy at 400x magnification. Datawere analysed statistically. The percentage of motility and viability both on pre-freezing and post-thawed process were significantly higher insemen diluted with TEY + 10% GFF compared to control (TEY), TEY + 5% GFF, and TEY + 15% GFF. Addition of 10% GFF into tris-egg yolkextender play role for maintaining the quality of pre-freezing and post-thawed Bali bull spermatozoa.

2019 ◽  
Vol 20 (1) ◽  
pp. 93
Author(s):  
Thomas Mata Hine ◽  
Kirenius Uly ◽  
Wilmientje Marlene Nalley ◽  
Heri Armadianto

Dimethyl Sulfoxide (DMSO) is one type of cryoprotectant which has a low molecular weight so that it is easier to enter cells when cryopreservation. The purpose of this study was to explore the optimal concentration of DMSO in modified coconut water (mCW) extender that were able to maintain frozen sperm quality of bali bulls. Semen was collected from two four-year old bali bulls by artificial vagina. Good quality semen diluted with mCW (young coconut water + 20% egg yolk + 7.5 % moringa leaf extract) and supplemented by 3, 5, or 7% DMSO. Semen was filled into 0.25 ml ministraw, and was incubated in a refrigerator at 5°C for four hours, frozen on the surface of liquid nitrogen for 10 minutes and then dipped into liquid nitrogen. The quality of post thawing sperm was measured 24 hours later by placing the ministraw of frozen semen into water at 37oC for 30 seconds. Data were analyzed by analysis of variance and continued with Duncan test. Postthawing observations showed that bali bulls sperm cryopreserved at 3% DMSO yielded higher motility and viability (p<0.05) i.e. 36 and 44.15%, than DMSO 5% i.e. 18 and 23.65%, and DMSO 7% i.e. 7 and 12.62%. The recovery rate of sperm cryopreserved at 3% DMSO was also higher (p<0.05) than DMSO 5 and 7%, successively 45.65, 23.06, and 8.86%. The results of this study concluded that the optimal concentration of DMSO in mCW diluent to maintain frozen sperm quality of bali bulls was 3%. 


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


2019 ◽  
Vol 64 (No. 03) ◽  
pp. 118-124 ◽  
Author(s):  
ME Inanc ◽  
S Gungor ◽  
C Ozturk ◽  
F Korkmaz ◽  
I Bastan ◽  
...  

The objective of this study was to determine effects of supplementing Tris-based semen extenders with either cholesterol-loaded cyclodextrin (CLC) or 7-dehydrocholesterol loaded cyclodextrin (7-DCLC) plus trehalose (T) for cryopreservation of ram semen. Semen was collected with an artificial vagina from five Merino rams (2–3 years of age) during the non-breeding season. Ejaculates were pooled, divided into eight equal portions, diluted with a standard Tris-based extender containing: no additive (control); T (50 mM); or T (50 mM) + 1.5, 2.5 or 3.5 mg of either 7-DCLC or CLC. Semen was chilled from 37°C to 4°C, placed in 0.25 ml French straws, held 5 cm above liquid nitrogen for 12 minutes, then plunged into liquid nitrogen. After thawing, a computer-aided semen analyzer system (CASA) was used to assess motility, whereas plasma membrane and acrosome integrity (PMAI) and high mitochondrial membrane potential (HMMP) were assessed with flow cytometry. Sperm supplemented with 2.5 mg and 3.5 mg CLC + T had the highest (P &lt; 0.05) total and progressive motility (65.2 ± 4.7 and 19.0 ± 1.0% respectively, mean ± SEM), albeit with no significant differences from sperm with 1.5 or 3.5 mg CLC + T. Sperm with 2.5 mg CLC + T had the highest (P &lt; 0.05) PMAI (59.3%; not different from 3.5 mg CLC + T) and highest (P &lt; 0.05) HMMP (64.6%; not different from 1.5 or 3.5 mg CLC + T). The lowest ALH value, 2.8 ± 0.3 µm was in the 2.5 mg 7-DCLC + T group; otherwise, there were no significant differences among groups for any other CASA end point. In conclusion, adding CLC + T to a tris-based extender optimized quality of frozen-thawed ram semen. Therefore, extenders including CLC + T have potential to improve quality of frozen-thawed ram sperm.


2020 ◽  
Vol 21 (12) ◽  
Author(s):  
KHAERUDDIN KHAERUDDIN ◽  
JUNAEDI JUNAEDI ◽  
HASTUTI HASTUTI

Abstract. Khaeruddin, Junaedi, Hastuti. 2020. Cryopreservation of Indonesian native chicken semen by using dimethyl sulfoxide and various level of ethylene glycol as cryoprotectants. Biodiversitas 21: 5718-5722. Imported purebred chickens are becoming more popular and a regular staple in Indonesia. Therefore, it is necessary to strengthen conservation efforts to preserve Indonesian chickens, one of which is by means of sperm cryopreservation. This study aimed to determine the effects of the addition of DMSO and different concentrations of ethylene glycol to a Ringer’s lactate egg yolk (RLY)-or coconut water egg yolk (CWY)-based extender on the quality of frozen-thawed Indonesian chicken sperm. This study was used nine Indonesian native roosters about 20 months of age. The semen extenders used in this study were RLY + DMSO 7%, RLY + ethylene glycol 3%, RLY + ethylene glycol 5%, RLY + ethylene glycol 7%, CWY + DMSO 7%, CWY + ethylene glycol 3%, CWY + ethylene glycol 5% and CWY + ethylene glycol 7%. Liquid semen was packaged in 0.25 mL straw, then cooled at 5oC for 2 hours, frozen at 5 cm above liquid nitrogen for 10 minutes, following stored in a liquid nitrogen container for 24 hours. The semen straws were thawed at 37oC for 30 seconds. Statistical analysis for multiple comparisons was performed as a completely randomized design with eight treatment levels and seven replications. The results showed that there were no differences in sperm motility, recovery rate, and abnormality between extenders after the freeze-thaw process. Whereas, RLY + DMSO 7% was the highest sperm viability.


2019 ◽  
Vol 20 (1) ◽  
pp. 20 ◽  
Author(s):  
ALoysius Marawali ◽  
Muhammad S. Abdullah ◽  
Jalaludin Jalaludin

The aim of this research was to know the effectiveness of guava filtrate supplementation in coconut water- egg yolk dilution on quality of liquid semen stored at 5oC of Bali cattle. Semen collected from a five year old Bali cattle using artificial vagina. Semen of good quality were kept in six tubes based on treatment then stored at 5oC. Treatments of the research were P0 : coconut water 80% + egg yolk 20% without guava filtrate; P1 : coconut water 80% + egg yolk 20% + 0.8% guava filtrate; P2 : coconut water 80% + egg yolk 20% + 0.9% guava filtrate; P3 : coconut water 80% + egg yolk 20% + 1.0 % guava filtrate; P4 : coconut water 80% + egg yolk 20% + 1.1 % guava filtrate and P5 : coconut water 80% + egg yolk 20% + 1.2 % guava filtrate. Each treatment was replicated 8 times making 48 experimental units. Results of the study showed that percentage mean of motility, viability, MPU, and TAU of spermatozoa after three days storage for P0 were : 42.20%, 41.85%, 39.08% and 40.90%; P1 : 50.40%, 53.89%, 52.99% and 54.67%; P2 : 54.67%, 56.97%, 54.51% and 54.36%; P3 : 17.00%, 29.96%, 29.64% and 29.64%; P4 : 23.38%, 24.64%, 21.06% and 24.45%Jurnal Veteriner Maret 2019 Vol. 20 No. 1 : 20 -29 pISSN: 1411-8327; eISSN: 2477-5665 DOI: 10.19087/jveteriner.2019.20.1.20 Terakreditasi Nasional, Dirjen Penguatan Riset dan Pengembangan, online pada http://ojs.unud.ac.id/index.php/jvet Kemenristek Dikti RI S.K. No. 36a/E/KPT/201621PENDAHULUAN Salah satu solusi yang dapat digunakan untuk pengembangan program Inseminasi buatan (IB) secara cepat dan mudah pada sapi bali adalah penggunaan semen cair. Penggunaan semen cair dapat meningkatkan kinerja IB pada sapi bali di Nusa Tenggara Timur (NTT). Keunggulan lain semen cair dapat diproduksi menggunakan bahan pengencer herbal berbasis bahan lokal dan peralatan yang sederhana serta mudah diperoleh dan tidak tergantung dengan persediaan nitrogen cair. Hasil akhir dari metabolisme spermatozoa adalah terbentuknya radikal bebas berupa derivat oksigen di antaranya adalah single1 oksigen (1O2), tripel1 oksigen (3O2), superokside anion (O2-), hidroksil radikal (OH) dan nitrit oxide (NO-) yang semuanya disebut radical oksigen species (ROS). Single1 oksigen dapat merusak ikatan rangkap pada asam lemak sehingga dapat menyebabkan kerusakan Deoxyribo Nuclead Acid (DNA) dan protein. Single1 oksigen bila bereaksi dengan asam amino histidin akan membentuk enzim yang dapat menyebabkan denaturasi protein. Kerusakan spermatozoa pada penyimpanan suhu 5%C akibat radikal bebas dan cold shock inilah merupakan penyebab utama disfungsi semen (Sharma et al., 2000). Oksidasi fosforilasi yang terganggu menyebabkan peningkatan radikal bebas dalam semen. Kadar radikal bebas yang terganggu menyebabkan peningkatan radikal bebas dalam semen. Kadar radikal bebas yang tinggi dalam sel dapat mengoksidasi lipid, protein dan DNA. Lipid membran plasma semen memiliki fosfolipid dengan kadar yang tinggi menyebabkan semen rentan terhadap radikal bebas (Sanoeka dan Kurpisz, 2004). Antioksidan bertindak mengikat asam lemak tak jenuh dan mencegah terjadinyareaksi berantai. Pada proses penyimpanan semen akan terjadi kerusakan membran plasma spermatozoa akibat terbentuknya perioksidasi lipid. Antioksidan-pemutus rantai seperti yang terkandung dalam jambu biji dapat menghambat perioksidasi lipid dalam membran melalui radical peroxyl (RO) dan alkoxyl (ROO) pengurai. Pengunaan jambu biji yang difilter dalam pengencer air kelapa kuning telur dapat menjaga kualitas spermatozoa (motilitas, keutuhan akrosom, viabilitas dan morfologi spermatozoa) semen cair sapi bali selama penyimpanan pada suhu 5%C. Dosis jambu biji yang difilter yang terbaik dalam pengencer air kelapa kuning telur, akan terbaik pula dalam mempertahankan kualitas spermatozoa sampai tujuan IB. Adapun tujuan penelitian ini adalah menguji berbagai level pemberian filtrat jambu biji (FJB) dalam pengencer air kelapa kuning telur terhadap motilitas, viabilitas, membran plasma utuh (MPU) dan tudung akrosom utuh (TAU) spermatozoa sapi bali yang disimpan pada suhu 5%C.METODE PENELITIAN Penelitian ini telah dilakukan di Laboratorium Reproduksi milik Yayasan Wiliams dan Laura yang berlokasi di Tilong, Desa Oelnasi, Kec. Kupang Tengah, Kab. Kupang, Nusa Tenggara Timur, dan berlangsung selama delapan bulan. Materi yang digunakan dalam penelitian ini adalah semen sapi bali yang ditampung dari satu ekor sapi bali jantan berumur lima tahun milik Yayasan Williams dan Laura yang telah dilatih, memiliki performans yang baik, dan organ reproduksi normal. Pakan yang diberikan adalah hijauan berupa rumput dan legum dan pemberian konsentrat secukupnya (dedak padi dan jagung giling).and P5 : 9%, 21.25%, 17.56% and 19.30%. Result of statistical analysis showed that there were a significant effect (P<0.05) between treatment on motility, viability, MPU and TAU of spermatozoa of Bali cattle till the third day of storage. It can be concluded that the supplementation of guava filtrate 0.9% in dilution of coconut water 80% - egg yolk 20% had been able to maintain motility, viability, MPU and TAU of spermatozoa of Bali cattle till the third day of storage at 5oC.


2006 ◽  
Vol 18 (2) ◽  
pp. 161
Author(s):  
A. Niasari-Naslaji ◽  
S. Mosaferi ◽  
A. A. Gharahdaghi ◽  
A. Abarghani ◽  
A. Ghanbari ◽  
...  

A Tris-based extender (SHOTOR diluent) has been developed for preserving Bactrian camel semen at 4�C (Niasari-Naslaji et al. 2005 Reprod. Fertil. Dev. 17, 198 (abstr.)). The present study investigated the possibility of utilizing the SHOTOR diluent for the cryopreservation of Bactrian camel semen. A modified bovine artificial vagina (Masaferi et al. 2005 Theriogeology 63, 92-101) was used to collect semen from three fertile bulls. The viscosity of the semen was reduced mechanically (Mosateri et al. 2005) and the homogenized semen was divided equally into two parts. Each part was sequentially diluted with either IMV buffers (Green buffer: first extender; White buffer: second extender; IMV, France) or SHOTOR diluents (without glycerol: first extender; with 12% glycerol: second extender). SHOTOR diluent consists of 2.6 g TIS, 1.35 g citric acid, 1.2 g glucose, and 0.9 g fructose in 100 mL of deionized water, with an osmolality of 330 mOsm/kg and pH of 6.9. All extenders had 20% egg yolk and antibiotics. The semen was diluted at the ratio of 1:1 with the first extender. The diluted semen was then cooled within 2 h to 4�C. At this temperature, the second extender was added at the same volume as the diluted semen in three steps with an equal volume, 10 min apart. After a 30-min equilibration time, beginning after addition of the last fraction of the second extender, the diluted semen was loaded into 0.5-mL straws at a concentration of 50 � 106 sperm per straw. The straws were maintained for 20 min at 4 cm above the liquid nitrogen surface, after which they were plunged into liquid nitrogen. The semen was thawed at 40�C water bath for 20 s. Progressive forward motility of spermatozoa was assessed at the time of dilution and immediately after thawing of the semen. The experiment was replicated four times. Data were analyzed using GLM procedure in SAS/STAT after arcsine transformation. At the time of dilution, there was no significant difference in progressive forward motility of spermatozoa between IMV buffers (51.8%) and SHOTOR diluent (61%; P > 0.05). However, after thawing, there was a significant decrease in progressive forward motility of spermatozoa in IMV buffers (4.2%) compared to SHOTOR diluent (29.9%, P < 0.05). In conclusion, in this experiment, SHOTOR diluent was more efficient for cryopreserving Bactrian camel semen than IMV extender. Shotor means camel in the Persian language.


2020 ◽  
Vol 4 (2) ◽  
pp. 1
Author(s):  
Zul Azhar ◽  
Fathurrahman Fathurrahman ◽  
Zul Mahadi Nata

<em>The role of water is very important in the growth process of the green deli honey guava plant, because this plant absolutely requires regular and sufficient water especially during the dry season. One of the obstacles for guava farmers is the problem of watering. Where honey guava has a moisture percentage ranging from 60% to 80% which must be maintained so that the quality of honey guava fruit is better. From these problems, an automatic watering device is designed based on soil moisture. Where this tool can also be monitored via android, LCD, and computer (PC) to make it easier for farmers to monitor. This study aims to determine the effect of soil moisture and assist farmers in controlling soil moisture in the cultivation of deli honey guava. The benefit of this research is to help farmers in an accurate and measurable process of watering plants. This research was conducted in Mancang Village, Binjai City. In this study, 2 honey guava trees were used to be tested using an automatic plant watering tool. One guava tree uses the manual method and the other is controlled using an automatic sprinkler. In this process the data is retrieved via a computer via USB with the PLX-DAQ software. The results showed that the soil moisture of the honey guava tree which was controlled using automatic sprinklers was more stable in maintaining soil moisture of 60% to 80% compared to the honey guava tree which was watered manually because the percentage of soil moisture did not match the plant's needs. Thus, the watering process using this tool is more controlled and measured in maintaining soil moisture in accordance with the needs of the green deli honey guava plant.</em>


Author(s):  
Abdullah Baharum ◽  
R. Iis Arifiantini ◽  
Tuty Laswardi Yusuf

The aims of this study were to investigate the freezing capability of Pasundan bull spermatozoa in Tris-egg yolk (TEY), Tris-soy (TS), and AndroMed® as diluents. Semen were collected twice a week from four Pasundan bulls aged 3-5 years old using an artificial vagina and evaluated macro- and microscopically. Semen had ≥70% sperm motility, ≥800x106/mL sperm concentration, and less than 20% sperm abnormalities were divided into three parts and each of them diluted with TEY, TS, or AndroMed®. After an equilibration step at 5°C for four hours, diluted semen were packaged in 0.25 mL straw, frozen in liquid nitrogen for ten minutes and kept in liquid nitrogen container until examination. Motility test on fresh, diluted, equilibrated, and after-thawed semen was done using Androvision®. The results showed that after thawing motility of sperm diluted in AndroMed® (58.64±0.72%) was higher than in TEY (49.45±1.22%) and TS (39.34±6.33%). Sperm motility of Pasundan bulls diluted in these three diluents reduced around 33.27±2.45% during freezing process.


2018 ◽  
Vol 35 (2) ◽  
pp. 150
Author(s):  
Naela Wanda Yusria Dalimunthe ◽  
M. Rosyid Ridlo ◽  
Agung Budiyanto

Buck slaugthering produce waste such as testicles including epididymis which contain fertile sperm. Utilization of cauda epididymis as the sources of sperm for producing goat frozen sperm was not reported yet. The aims of this study were improving the frozen-thawed sperm using stabilization and multistep methods which recovered from the waste of buck slaughtering as the source of sperma. Cauda epididymis spermatozoa which was washed then diluted using extender 1 (Tris-citrate-antibiotics) and extender 2 (extender 1- glycerol-egg yolk). The extender 2 addition was performed by single or multistep methods then freezed. Modification in the pre freezing proces were performed by comparing the conventional equilibration and stabilization methods. The sperm suspension was incubated in 4°C for 2 hours after filling-sealing into straws on the equilibration group whether the stabilization group was cooled in tube 15 mL. All cooled straws from both groups were placed 4 cm horizontally on liquid nitrogen surface for 10 minutes and then plunged into liquid nitrogen for storage. The evaluation of motility parameters such as pattern of the movement and motility percentation were done followed the standard methodology. The student t-test, correlation and one-way ANOVA were used for data analysis with P<0.05. The results showed that multistep dilution method could increase the motility (25.0 ± 1.8 %) compared with single step (18.3 ± 1.7 %). Pre freezing method with stabilization also resulted higher motility (24.2 ± 2.0 %) than equilibration method (17.5 ± 2.8 %). The pattern of the movement were not different between all methods and its combination. The multistep dilution method and stabilization cooling method as well as its combination seems could increase the quality of frozen-thawed cauda epididymis spermatpzoa of local buck.


Author(s):  
Khairuddin Khairuddin ◽  
Muhammad Erik Kurniawan ◽  
Soman Soman

The aim of this study was to determine the type and the best concentration of egg yolk in maintaining the quality of kampung rooster spermatozoa during cryopreservation. This study used a completely randomized factorial pattern design with the first factor was the type of egg yolk (purebred chicken, kampung chicken, duck, and quail) and the second factor was the concentration of egg yolk (5%, 10%, and 15%). Semen was collected from twelve kampung roosters using massage method. Immediately after collection, the semen was evaluated macroscopically and microscopically. Semen with more than 70% motility was used in this study. The semen was diluted, packed in a ministraw, equilibrated, and frozen using liquid nitrogen vapor and stored in a liquid nitrogen container for 24 hours. Observation of spermatozoa motility was carried out in fresh semen, diluted semen, after equilibration and after thawing with four replications. The results showed that the type of egg yolk treatment had no effect (P0.05) on the recovery rate and motility of spermatozoa before and after cryopreservation, but egg yolk concentration had a highly significant effect (P0.01) on the quality of spermatozoa. Egg yolks in 10-15% concentration had spermatozoa motility and recovery rate higher than egg yolk with 5% concentration. In conclusion, purebred chicken egg yolk, kampung chicken egg yolk, duck egg yolk, and quail egg yolk each in diluent can be used to maintain the quality of kampung rooster spermatozoa at a concentration of 10-15% during cryopreservation.


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