scholarly journals Pemijahan Ikan Endemik Pelangi Arfak (Melanotaenia Arfakensis) Secara Alami

2021 ◽  
Vol 2 ◽  
pp. 39-43
Author(s):  
Vicky Arnold Mofu ◽  
Bayu Pranata ◽  
Marhan Manaf
Keyword(s):  
Survival Rate ◽  
Larval Survival ◽  
Hatching Rate ◽  
West Papua ◽  
The Status ◽  
Endemic Fish ◽  
Lake Systems

Arfak rainbowfish (Melanotaenia arfakensis) is one of the endemic fish of West Papua that can be found in river and lake systems in the Manokwari Regency. Currently, the status of the arfak rainbowfish is nearly extinct. Therefore, this study aimed to know the hatching rate and survival of arfak rainbowfish larvae that were naturally spawned. The results showed that the hatchability of arfak rainbowfish eggs reached 96.9% in A1 aquarium and 96.4% in A2 aquarium. The survival rate of arfak rainbowfish larvae in the A1 aquarium was 67.5%, while the A2 aquarium was 58.7%. The larval survival rate was relatively low. Therefore, further research is needed to increase the survival rate of arfak rainbowfish larvae. 

scholarly journals REPRODUKSI IKAN BAUNG (Hemibagrus nemurus) DENGAN PERLAKUAN DOSIS HORMON GnRH-a BERBEDA

2018 ◽  
Vol 13 (3) ◽  
pp. 213
Author(s):  
Jojo Subagja ◽  
Vitas Atmadi Prakoso
Keyword(s):  
Body Weight ◽  
Survival Rate ◽  
Larval Survival ◽  
Fertilization Rate ◽  
Fertility Rate ◽  
Time Interval ◽  
Hatching Rate ◽  
Good Prospect ◽  
Hormonal Manipulation ◽  
Different Doses

Ikan baung (Hemibagrus nemurus) memiliki prospek bagus untuk dikembangkan sebagai komoditas budidaya di Indonesia karena citarasa daging yang enak banyak diminati konsumen. Dewasa ini, peningkatan produksi benih dapat dilakukan melalui pemijahan buatan dengan manipulasi hormonal. Penelitian yang dilakukan bertujuan untuk mengevaluasi produksi induk ikan baung melalui pemijahan buatan dengan dosis penyuntikan GnRH-a berbeda. Penelitian dilakukan di Instalasi Riset Plasma Nutfah Perikanan Air Tawar, Cijeruk, Bogor pada bulan November 2017. Induk betina ikan baung yang digunakan berukuran 465,0 ± 71,8 g; dan induk jantan 426,3 ± 46,8 g. Induksi dilakukan dengan penyuntikan hormon GnRH-a dengan tiga dosis yang berbeda pada induk betina (0,3; 0,5; dan 0,7 mL/kg bobot badan), dan ikan jantan dengan dosis 0,4 mL/kg bobot badan. Masing-masing perlakuan terdiri atas tiga ekor induk betina sebagai ulangan. Hormon disuntikkan secara intramuskular, diberikan dua kali penyuntikan 35% dari dosis total pada penyuntikan pertama, dan 65% diberikan pada penyuntikan kedua, dengan interval waktu penyuntikan delapan jam. Parameter yang diamati yaitu jumlah telur ovulasi, derajat pembuahan, derajat penetasan, dan sintasan larva selama tujuh hari. Hasil penelitian menunjukkan bahwa dosis penyuntikan hormon GnRH-a 0,5 mL/kg pada induk ikan baung saat proses pemijahan buatan menghasilkan derajat penetasan yang lebih tinggi dibandingkan dosis 0,3 dan 0,7 mL/kg; serta sintasan larva yang lebih baik dibandingkan dosis 0,7 mL/kg. Sedangkan nilai jumlah telur yang berhasil ovulasi dan derajat pembuahan yang relatif lebih baik ditemukan pada perlakuan dosis 0,7 mL/kg dibandingkan dengan dosis 0,3 dan 0,5 mL/kg.Asian redtail catfish (Hemibagrus nemurus) has a good prospect to be developed as aquaculture commodity in Indonesia. It was proved by the demand of consumers regarding to its delicious flesh taste. Recently, efforts to increase seedling production could be implemented through artificial spawning with hormonal manipulation. This study was conducted to evaluate the production of broodstocks through artificial spawning with different doses of GnRH-a. Study was conducted at Research Institute for Freshwater Fisheries Germplasm, Cijeruk, Bogor in November, 2017. The broodstock used in this study was 465.0 ± 71.8 g for females, and 426.3 ± 46.8 g for males. Induction was performed by injection of GnRH-a hormone with three different doses on female broodstocks (0.3, 0.5 and 0.7 mL/kg of body weight), and male broodstocks with a dose of 0.4 mL/kg of body weight. Each treatment consisted of three females as replications. The hormone was injected intramuscularly, given twice injection: 35% of the total dose is given at the first injection, and the remaining 65% was given at the second injection, with an injection time interval of eight hours. The results showed the observed parameters of number of the ovulated eggs, fertilization rate, hatching rate, and larval survival for seven days, treatment dosage of 0.5 mL/kg showed better results (P<0.05) than the dosage of 0.3 mL/kg and 0.7 mL/kg. Parameters observed were number of ovulated eggs, fertility rate, hatching rate, and survival rate for seven days. The results showed that the dosage of 0.5 mL/kg GnRH-a injection to the broodstock during artificial spawning resulted in higher hatching rate compared to 0.3 and 0.7 mL/kg and better larval survival rate than the dosage of 0.7 mL/kg. Meanwhile, the number of ovulated eggs and fertility rate was relatively better in the treatment of 0.7 mL/kg compared with the dosage of 0.3 and 0.5 mL/kg.

Egg protein bound amino acid content and embryo/larva success in common dentex (Dentex dentex), a marine pelagophil teleost

2013 ◽  
Vol 63 (1) ◽  
pp. 59-75 ◽  
Author(s):  
Seyed-Mohammadreza Samaee ◽  
Eleni Mente ◽  
Alicia Estévez
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Survival Rate ◽  
Acid Content ◽  
Amino Acid Content ◽  
Larval Survival ◽  
Hatching Rate ◽  
Egg Protein ◽  
In Captivity ◽  
Dentex Dentex

This study represents the initial attempt to evaluate the effect of egg protein-bound amino acid (i.e., vitellogenin-derived PAAs) content on embryo/larva success in Dentex dentex. Viable eggs were obtained from a single female (i.e., intra-individual analysis) in captivity. Selected embryo/larva viability parameters (the egg floating rate, the hatching rate, and the larval survival rate at days 0 to 5 post-hatch) were determined. The egg batches were categorised in three groups (high-, medium-, and low-quality batches) based on the magnitude of their hatching rates. The protein-bound amino acid content of biometrically similar egg batches of varying quality was analysed. The content values for 12 protein-bound amino acids (i.e., essential PAAs: Thr, Met, Arg, Ile, Val, Phe, His; non-essential PAAs: Ala, Glu, Gly, Asp, and Ser) were significantly correlated with the embryo/larva viability parameters in an analysis performed in terms of their ratios. The egg protein-bound amino acid content was significantly correlated with the dentex larval survival rate at 3 and 5 days post-hatch, a critical period of survival that is problematic during the propagation of this species in captivity. The results of this study also offer statistical evidence for the effects of the interrelationships of the egg protein-bound amino acids on the relationships between the quantitative composition of the protein-bound amino acids and the viability parameters.

scholarly journals PENGARUH DOSIS OVAPRIM TERHADAP LAMA WAKTU PEMIJAHAN, DAYA TETAS TELUR DAN KELANGSUNGAN HIDUP LARVA IKAN BIAWAN (HELOSTOMA TEMMINCKI)

2016 ◽  
Vol 4 (1) ◽  
Author(s):  
Eka Indah Raharjo ◽  
Rachimi . ◽  
Holidan .
Keyword(s):  
Body Weight ◽  
Survival Rate ◽  
Fish Farming ◽  
Larval Survival ◽  
Hatching Rate ◽  
Spawning Period ◽  
Egg Hatchability ◽  
Quality Of Water ◽  
Randomized Design ◽  
Hatching Eggs

ABSTRAK Penelitian ini telah dilaksanakan di Balai Budidaya Ikan Sentral (BBIS) Anjongan, Kabupaten Mempawah Provinsi Kalimantan Barat. Waktu pelaksanaannya 14 Hari, meliputi 3 hari persiapan alat dan bahan dan 11 hari pengamatan penelitian. Penelitian ini bertujuan untuk menentukan dosis hormon Ovaprim yang dapat menghasilkan lama waktu pemijahan, daya tetas telur dan sintasan hidup pada larva ikan biawan. Metode penelitian yang digunakan adalah metode eksperimen. Rancangan yang digunakan adalah Rancangan Acak Lengkap (RAL)  dengan 4 perlakuan dan 3 ulangan. Variabel pengamatan meliputi waktu ovulasi, daya tetas telur (hatcing rate), kelangsungan hidup larva. Berdasarkan hasil penelitian menunjukkan bahwa pemberian dosis ovaprim yang berbeda berpengaruh nyata terhadap waktu ovulasi dan daya tetas telur ikan dan kelangsungan hidup ikan biawan. Nilai waktu ovulasi tercepat terdapat pada perlakuan B (dosis ovaprim 0,6 ml/kg bobot tubuh) dengan waktu ovulasi (8,79) jam.  Nilai daya tetas (hatching rate) tertinggi terdapat pada perlakuan B (dosis ovaprim 0,6 ml/kg bobot tubuh) sebesar 86,53 %, Nilai kelangsungan hidup larva biawan tertinggi terdapat pada perlakuan B (dosis ovaprim 0,6 ml/kg bobot tubuh) sebesar 86.51 %. Kualitas air selama pemijahan dan penetasan telur selama penelitian diperoleh suhu 27-29°C  pH berkisar antara 6,5-7. Oksigen terlarut adalah 5-6 ppm. Kata Kunci: dosis Ovaprim, lama waktu pemijahan, hatching rate, kelangsungan hidup, Helostoma temmincki.  ABSTRACTThis study was conducted at the Fish Farming Center of Balai Budidaya Ikan Sentral Anjongan, Kabupaten Mempawah, West Kalimantan Province. It took 14 days for preparation (3 days) and observation (11 days). This study aimed at determining  the dose of Ovaprime hormones that produced spawning period, and egg hatchability and survival rate of Biawan larvae The method used is an experimental method. Using exprimental methodand completely randomized design (CRD), this study used  4 treatments and 3 repetitions. The variabels employed in observation phase were ovulation time, egg hatchability rate, and larval survival. The study revealed that different ovaprim  dosing significantly affectedthe ovulation time and the egg hatchability and  survival of Biawan. The fastest value of ovulation time was found in treatment B (ovaprim dose of 0.6 ml / kg body weight) at ovulation time (8.79) hours.  The highest hatching rate was found in treatment B (ovaprim dose of 0.6 ml / kg body weight) amounted to 86.53%. Also, the highest value of Biawan larval survival was found in treatment B (ovaprim dose of 0.6 ml / kg body weight) amounted to 86.51%. In addition,  the quality of water for spawning and hatching eggs was at  a temperature of 27-29 ° C pH range between 6.5-7, and  the dissolved oxygen was 5-6 ppm. Keywords: Ovaprim dose,  spawning period, hatching rate, survival rate, Helostoma temmincki.

scholarly journals Batesian mimicry has evolved with deleterious effects of the pleiotropic gene doublesex

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Shinya Komata ◽  
Tasuku Kitamura ◽  
Haruhiko Fujiwara
Keyword(s):  
Survival Rate ◽  
Survival Rates ◽  
Cost Benefit ◽  
Larval Survival ◽  
Batesian Mimicry ◽  
Hatching Rate ◽  
Swallowtail Butterfly ◽  
Adult Lifespan ◽  
Pleiotropic Gene ◽  
Papilio Polytes

AbstractDimorphic female-limited Batesian mimicry in the swallowtail butterfly Papilio polytes is regulated by the supergene locus H, harbouring the mimetic (H) and non-mimetic (h) doublesex (dsx) gene. In the present study, we demonstrated that dsx-H negatively affects the number of eggs laid, hatching rate, larval survival rate, and adult lifespan. When crossed with hh males, the number of eggs laid of mimetic females (genotype HH) was lower than that of non-mimetic females (hh). Moreover, hh and Hh females laid fewer eggs when crossed with HH males. The hatching and larval survival rates were lower when both female and male parents harboured dsx-H. The adult lifespan of HH females was shorter than that of hh females, while it was similar in males regardless of the genotype. These findings suggest the presence of a cost–benefit balance of Batesian mimicry, which is evolved to avoid predation but is accompanied by physiological deficits, in this species.

EFFECTIVENESS Daphnia sp. WHICH IS PELET FEEDING TO THE GROWTH OF SEMAH’S LARVAE Tor douronensis (Valenciennes, 1842)

2020 ◽  
Vol 5 (2) ◽  
pp. 23-27
Author(s):  
Dina Tri Madya Ningsih ◽  
Siti Hudaidah ◽  
Mas Tri Djoko Sunarno
Keyword(s):  
Survival Rate ◽  
Fish Larvae ◽  
Weight Growth ◽  
Randomized Design ◽  
Test Parameters ◽  
Absolute Growth ◽  
Absolute Length ◽  
Effective Doses ◽  
Endemic Fish ◽  
Completely Randomized Design

The semah fish (Tor douronensis) is an Indonesian endemic fish that has a long growth, which takes 4 years to grow from larvae to broodstock. This study aims know of Daphnia sp. pelet fed with effective doses to increase the growth of semah’s fish larvae. The study was conducted from March 25 to April 13, 2020 at the Instalasi Riset Plasma Nutfah, Cijeruk, Bogor, West Java. The experimental design used completely randomized design (CRD) 4 treatments and 3 replications, namely treatment A (Daphnia sp.+pelet feed 0 gr/ L), treatment B (Daphnia sp.+ pelet feed 1 gr / L), treatment C (Daphnia sp. + pelet feed 2 gr / L), treatment D (Daphnia sp. + pelet feed 3 gr / L). The larvae of 9 days in age were randomly stocked into 12 aquariums measuring 60x30x35 cm with a density of 50 fish/aquarium and fed in the form of Daphnia sp. enriched pelet feed according to the treatment of 50 ind/larvae by giving 3 times a day which is maintained for 20 days. The test parameters measured are absolute growth in length and weight, survival rate, and water quality. Based on the results of statistical analysis (Anova), the results showed that the treatment D gave the highest influence on the growth of semah’s fish larvae (P <0.05) with an absolute weight growth value of 85,3 mg, absolute length growth of 10.6 mm, and survival rate 99.33%.

scholarly journals ANALISIS KARAKTER REPRODUKSI DAN PERFORMA BENIH HIBRID IKAN NILA PANDU F6 DENGAN IKAN NILA NILASA (Oreochromis niloticus) Analysis of the Character Reproduction and Seed Hybrid Performance 0f Tilapia Fish F6 Pandu with Nilasa Tilapia (Oreochromis niloticus)

2017 ◽  
Vol 12 (1) ◽  
pp. 19
Author(s):  
Dio Patria Yustysi ◽  
Fajar Basuki ◽  
Titik Susilowati ◽  
Tristiana Yuniarti
Keyword(s):  
Water Quality ◽  
Growth Rate ◽  
Survival Rate ◽  
Specific Growth Rate ◽  
Oreochromis Niloticus ◽  
Egg Size ◽  
Specific Growth ◽  
Fish Farming ◽  
Hatching Rate ◽  
Feed Conversion

ABSTRAK Penelitian ini bertujuan untuk mengetahui pengaruh dari hibridisasi ikan nila Pandu F6 dengan nila Nilasa terhadap karakter reproduksinya dan performa benih yang dihasilkan. Penelitian ini dilakukan dengan metode eksperimental menggunakan Rancangan Acak Lengkap (RAL) dengan 4 perlakuan dan 3 kali ulangan. Perlakuan dalam penelitian ini: perlakuan A (Pandu F6 ♀><Pandu F6 ♂), B (Nilasa ♀><Nilasa ♂), C (Pandu F6 ♀>< Nilasa ♂), dan D (Nilasa ♀><Pandu F6 ♂). Data yang diamati meliputi fekunditas, daya tetas telur, diameter dan bobot telur, panjang dan bobot larva TL, panjang dan bobot larva lepas kuning telur, kelulushidupan, laju pertumbuhan spesifik, konversi pakan dan kualitas air. Hasil penelitian menunjukan nilai terbaik pada fekunditas sebesar 1191,67 ± 239,45 butir/200 g, daya tetas telur sebesar 80,93 ± 3,90%, kelulushidupan sebesar 81,55 ± 7,54%, laju pertumbuhan spesifik sebesar 7,26 ± 0,25, konversi pakan 0,54 ± 0,02 didapatkan pada perlakuan C, akan tetapi untuk diameter dan bobot telur, panjang dan bobot larva Kuning Telur, dan panjang dan bobot larva lepas kuning telur tidak berbeda nyata. Hasil tersebut menunjukan bahwa hibridisasi (perlakuan C) memberikan pengaruh yang nyata (P < 0,05) terhadap fekunditas, daya tetas telur, kelulushidupan, laju pertumbuhan spesifik, dan konversi pakan tetapi tidak berpengaruh terhadap diameter dan bobot telur, panjang dan bobot larva, dan panjang dan bobot larva lepas kuning telur. Kata kunci: Hibridisasi, Ikan Nila, Karakter Reproduksi, Performa Benih  ABSTRACT This research aims to know the influence of hybridization tilapia fish Pandu F6 with nila Nilasa against the character of the reproduction and seed performance. This research was conducted with the experimental method using Random Design complete (RAL) with 4 treatments and three replicates. The treatments in this study: the treatment A (Pandu F6 F6 ♀ >< Pandu ♂), B (Nilasa ♀>< Nilasa ♂), C (Pandu F6 ♀>< Nilasa ♂), and D (Nilasa ♀ >< Pandu F6 ♂). The observed data covering fecundity, hatching rate, egg size, yolk sack larva length and weight, length and weight of the egg yolk off larvae, survival rate, specific growth rate, feed conversion rate, and water quality. The results showed the best value on the fecundity of 1191.67±239.45 eggs/200 g, Hatching rate 80.93 ± 3.90%, Survival rate of 81.55 ± 7.54%, specific growth rate of 7.26 ± 0.25, conversion fodder 0.54 ± 0.02 obtained at the treatment C, but for the diameter and weight of egg, larval length and weight TL, and the length and weight of the yolk egg off larvae for each treatment do not differ markedly. The results showed that hybridization (treatment C) gives a real influence (P < 0.05) of fecundity,hatching rate, survival rate but not with the egg size, larva weight and length, and the length and weight of larvae off yolk. Water quality on the spawning, hatching eggs and larvae found on the maintenance of a decent range for tilapia fish farming Keywords: hybridization, tilapia fish, character of the reproduction, seed performance 

scholarly journals Daya Tetas Telur dan Sintasan Larva Dari Hasil Penambahan Madu pada Bahan Pengencer Sperma Ikan Nila (Oreochromis niloticus)

2019 ◽  
Vol 3 (1) ◽  
Author(s):  
Yunus Ayer ◽  
Joppy Mudeng ◽  
Hengky Sinjal
Keyword(s):  
Experimental Design ◽  
Oreochromis Niloticus ◽  
Nile Tilapia ◽  
Larval Survival ◽  
Hatching Rate ◽  
Dilution Ratio ◽  
Egg Hatching ◽  
Research Results ◽  
Spermatozoa Motility ◽  
Randomized Design

The objective of research was to determine the concentration of honey in sperm dilution  for improving hatching rate of egg and survival of nile tilapia larvae (Oreochromis niloticus).The number of fish used was five pairs broodstock. Dilution substance was NaCl and honey.  Observations were conducted on spermatozoa motility, fertility and egg hatching rate. Experimental design used was complete randomized design. Sperm dilution substance was prepare by disolving honey  (0 mL; 0,60 mL; 0,65 mL dan 0,70 mL) in 100 mL; 99,40 mL; 99,35 mL and  99,30 mL NaCl respectively.  Each dilution was homogenized using aerator for 15 minutes. Fertilization was done 12 hours after ovulation.  Egg hatching rate was observed after ovulation. Research results showed the use of honey had significant effet on egg hatching rate, but not on larval survival.  Treatment D (0,70 mL honey in 99,30 mL NaCl) had the highest hatching rate (77.33%). Dilution ratio 1:60 was the best indicated by spermatozoa motility 96.66%, fertility 71.65, hatching rate 70% and larval survival 81.67%   Keywords : honey, egg hatching rate, larval survival, Oroechromnis niloticus

scholarly journals Effect of feeding management of broodstock on breeding performance of bata (Labeo bata)

2016 ◽  
Vol 1 (3) ◽  
pp. 553-568 ◽  
Author(s):  
Md Habibur Rahman ◽  
Md Anisur Rahman ◽  
Md Mer Mosharraf Hossain ◽  
Syeda Maksuda Yeasmin ◽  
Abdulla Al Asif
Keyword(s):  
Methylene Blue ◽  
Survival Rate ◽  
Lipid Level ◽  
Gonadosomatic Index ◽  
Dietary Factors ◽  
Hatching Rate ◽  
Breeding Performance ◽  
Fish Hatchery ◽  
Labeo Bata ◽  
Significant Difference

Labeo bata is one of the important minor carps in Bangladesh with great demand as good table fish. The study was conducted to observe the breeding performance at different management practices in Mafatema, Rupali, Modhumoti and Anan fish hatchery and disinfection treatments of water, eggs and fry in Jessore, Bangladesh during 10 March 2014 to 15 May 2015. An improvement in broodstock nutrition and feeding has been shown to greatly improve seed production. Protein and lipid percentage of broodstock diet have been identified as major dietary factors. Protein level was 24.77%, 23.47%, 18.08%, 17.78% and lipid level was 11.07%, 9.50%, 7.74%, 8.14% in Mafatema, Rupali, Modhumoti and Anan fish hatchery respectively. Three concentrations of four chemical-formalin (10, 20, 30 mg/L), malachite green (1, 3, 5 mg/L), NaCl (1, 2, 3 g/L) and methylene blue ( 1, 3, 5 mg/L) treatment regimes and a control were compared for efficacy in treating L. bata eggs to prevent fungus and bacterial infection and improve hatch and survival rate of fry. Highest correlation value between absolute fecundity and body weight (r=.938, p<.05) and total length (r=.891, p<.05) and gonadosomatic index (26.2%) were found in Mafatema fish hatchery among four experimental hatcheries at 24.77% protein and 11.07% lipid level. Better fertilization rate (84.2±5.17%) and hatching rate (82.0±4.30%) were found in Mafatema and Rupali fish hatchery respectively that has significant difference (P<0.05) from that of Modhumoti and Anan fish hatchery at higher protein and lipid level. Lowest deformity rate (6.05+2.65) was observed in Mafatema fish hatchery that was significantly different (P<0.05) from that of Modhumoti fish hatchery. In case of disinfection treatment, methylene blue at 1mg/L bath treatment daily for 4 days showed significantly higher hatching rate (92.33±3.51%) and survival rate (94.33±4.73%).Asian J. Med. Biol. Res. December 2015, 1(3): 553-568

52 EFFECT OF CARBOXYLATED POLY-L-LYSINE ON THE POST-THAW VIABILITY OF IN VITRO-PRODUCED BOVINE BLASTOCYST

2015 ◽  
Vol 27 (1) ◽  
pp. 119
Author(s):  
A. N. Ha ◽  
K. L. Lee ◽  
Md. Fakruzzaman ◽  
S. S. Kim ◽  
P. R. Park ◽  
...  
Keyword(s):  
Survival Rate ◽  
Dimethyl Sulfoxide ◽  
Exposure Time ◽  
High Efficiency ◽  
Apoptotic Cell ◽  
Osmotic Shock ◽  
Hatching Rate ◽  
Cell Numbers ◽  
Bovine Blastocyst

Recently, there has been development of an antifreeze polyamino acid (carboxylated poly-l-lysine: PLL) as new cryoprotectants (CPA). This compound counts as amphoteric macromolecular cationic and anionic substituents (polyampholyte) by chemically modifying poly-lysine. In addition, PLL is highly safe and frequently used as a food additive in substitution for other CPA. Other common CPA have high toxicity and caused physiological damage. In vitro-produced blastocysts were randomly assigned into 3 groups: (1) vitrified embryos with PLL vitrification solution [PLL-vit-1: 15% PLL + 15% ethylene glycol (EG); PLL-vit-2: 30% PLL + 30% EG + 0.5 M sucrose], (2) vitrified embryos with Vajta et al. solution (Conv-vit-1: 7.5% dimethyl sulfoxide + 7.5% EG; Conv-vit-2: 16.5% dimethyl sulfoxide + 16.5% EG + 0.5 M sucrose), and (3) nonvitrified blastocysts (control). All embryos were frozen by droplet vitrification method. First, the PLL-vitrified embryos were exposed to 5, 10, and 15 min in the PLL-vit-1 and then putted for 30 to about 60 s in the PLL-vit-2. Then, we compared with each group regarding exposure time of Vit-1. Post-thaw survival rate of each exposure time did not significantly differ among the 3 groups (100 v. 100 v. 100%). However, hatching rate of the 10-min exposure group was higher than that of 5- and 15-min groups (75.0 v. 25.0 v. 66.7; P < 0.05). Therefore, we confirmed that exposure time of Vit-1 was exposed for a minimum of 10 min. The post-thawed survival rate of each vitrification method was not significantly different between PLL-vit and Conv-vit groups (97.7 v. 86.4%). The total cell numbers of blastocyst did not significantly differ among groups. However, the apoptotic cell numbers of blastocyst was significantly different between the control and Conv-vit groups (0.4 ± 0.6 v. 4.4 ± 3.9; P < 0.05) but was not different in control v. PLL-vit (0.4 ± 0.6 v. 2.1 ± 2.4) and Conv-vit v. PLL-vit (4.4 ± 3.9 v. 2.1 ± 2.4). In conclusion, PLL-vit for bovine blastocyst could reduce toxicity and osmotic shock and showed high efficiency on the quality of post-thawed bovine blastocysts compared with that of Conv-vit.This work was partly supported by a grant from the Next-Generation BioGreen 21 Program (No. PJ009587022014), IPET (Grant No. 112020-3), and a scholarship from the BK21 plus program. A-Na Ha, Kyeong-Lim Lee, and Md. Fakruzzaman were supported by BK21 plus fellowships at Gyeongsang National University, Republic of Korea.

47 A New Device and Method for Successful Vitrification of In Vitro-Produced Ovine Embryos

2018 ◽  
Vol 30 (1) ◽  
pp. 163
Author(s):  
S. Ledda ◽  
J. M. Kelly ◽  
S. K. Walker ◽  
Y. Natan ◽  
A. Arav
Keyword(s):  
Survival Rate ◽  
Embryo Transfer ◽  
Blastocyst Stage ◽  
Embryo Cryopreservation ◽  
Control Group ◽  
Hatching Rate ◽  
High Survival ◽  
Embryo Survival ◽  
New Device

To advance the use of embryo vitrification technology in veterinary practice, we developed a system in which embryo vitrification, warming, and dilution can be performed within a straw. An in-straw embryo cryopreservation method reduces the need for equipment and technical skills and can facilitate direct embryo transfer to the uterus. This study proposes the use of a new device named “Sarah” that is designed to permit all in-straw embryo cryopreservation procedures. Ovine in vitro-produced (IVP) embryos were vitrified at either early blastocyst stage (EB, n = 65, 6 days post-IVF) or fully expanded blastocyst stage (FB, n = 168, 7 days post-IVF). The vitrification procedure using Sarah constituted a 0.25-mL straw with a capsule having 50-µm pores inserted at one end. Embryos at each stage (EB and FB) were divided into 2 subgroups and vitrified by 1 of 2 methods: (1) multi-step (MS) group-a straw containing 2 embryos was sequentially loaded vertically into 1.5-mL tubes containing 6 different vitrification solutions: 10, 20, 40, 60, 80, or 100% ES (with 100% ES being 7.5% DMSO +7.5% EG + 20% FCS in TCM-199; 90 s each step) followed by 30 s each in 75 and 100% VS (100% VS being 18% DMSO +18% EG + 0.5 M trehalose + BSA in TCM-199); and (2) two-step (TS) group-the straw (2 embryos/straw) was loaded with 100% of ES (5 min), followed by 100% VS solution for 30 s. For both methods, at the end of the preparation steps, the straws were plunged directly into liquid N2. Non-vitrified embryos were maintained in in vitro culture as a control group (n = 102). The warming procedure consisted of placing the straws directly into 5-mL tubes containing 100, 50, 25% WS (WS = 1 M sucrose in TCM-199+ 20% FCS) at 38.6°C (for first solution) and at room temperature for all the rest (5 min each), before being placed into the holding medium. Embryos were recovered from the straws, incubated at 38.6 C in 5% CO2 in air in TCM 199 + 5% FCS, and evaluated for blastocoel re-expansion, embryo survival, and hatching rate at 2, 14, 48 h post-warming. Blastocyst re-expansion (2 h) after warming increased as the developmental stage progressed and was not affected by the vitrification method. In fact, it was significantly (P < 0.05) higher for FB vitrified in the MS and TS methods (77.90% and 71.25%, respectively) compared with the EB method (62.5% and 48.50%, respectively). At 24 h, survival rate of vitrified FB was significantly higher (P < 0.05) in the MS system (95.35%) compared with those in TS (86.25%). Survival rates of FB embryos for both methods (MS and TS) were significantly higher (P < 0.001) than EB embryos vitrified in MS (56.25%) and TS (56.55) methods. After 48 h of culture, the hatching rate for FB vitrified in the MS system (87.21%) was comparable with TS (77.5%) and control (85.3%) groups but significantly higher (P < 0.001) than vitrified EB in MS (43.75%) and TS (36.36%). In conclusion, we showed that a high survival rate of IVP embryos can be achieved by this new in-straw vitrification and warming device (“Sarah”), with hatching rates in vitro comparable with that of control fresh embryos. This method has the potential for use in direct embryo transfer in field conditions.

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