• Journal of Embryo Transfer
• /
• v.33 no.3
• /
• pp.169-175
• /
• 2018

It has been claimed that artificial insemination (AI) of cows with frozen-thawed semen treated with commercially produced kits, Wholemom (in favour of female gender) increases the birth chance of calves with desired sex ratio by approximately 85% without decrease of pregnancy rates. Hence, this study was conducted to investigate the efficacy of wholemom kits as combined with frozen-thawed bovine semen during in vitro fertilization on the in vitro fertilization and developmental efficiency and sex ratios such as some reproductive parameters in bovine. For this, 1,737 oocytes were in vitro fertilized and developed. Agglutination effects on bovine after treatment of Wholemom kit were observed by time passage and dose respectively. To determine sex of embryos, Bovine embryo Y-specific gene primers(ConEY) and Bovine specific universal primer(ConBV) were used as multiple PCR method. Fertilization rate of wholemom-treated group was significantly lower than its of control group[66.9% (1,156/1,737) in Wholemom-treated group; 75.0% (610/813) in control group]. However, developmental rate after fertilization of both wholemom-treated and control groups were not significantly different [26.1% (404/1,156) in Wholemom-treated group; 27.4% (224/610) in control group]. Sex ratio of in vitro fertilized embryo with frozen-thawed semen treated with wholemom kit was determined by multi PCR. Female ratio in wholemom-treated group [85.4% (173/201)] was significantly higher than its of control group [47.2% (66/141)]. In conclusion, wholemom treatments of semen used in the in vitro fertilization and development of bovine oocytes provided increase in female ratio with decrease of fertilization rate.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.12
• /
• pp.1652-1656
• /
• 2004

The aim of this study was to obtain mature ova or embryos at a single cell stage, which can be used in avian transgenesis and nuclear transfer through multiple ovulations, in vitro fertilization and culture. Chicken anterior pituitary extract (CAPE) or acetone-dried chicken anterior pituitary extract (ACAPE) was used to induce multiple ovulations in hens pretreated with pregnant mare' serum gonadotrophin (PMSG). In vitro fertilization of the multiple ovulated ova was performed by inseminating sperm onto the germinal disks in m-Ringer' solution and incubating the ova at 41$^{\circ}C$, 5% $CO_2$ for 10 h in DME-F12 medium containing 20% liquid albumen. The in vitro fertilization process was observed using an environmental scanning electron microscope. When normal laying hens (white Leghorn) were administered daily with PMSG (100 IU), egg laying ceased in most hens within 3 to 8 days. Ovulation began to occur about 7.5 h after injection of CAPE and ACAPE. The number of ovulated ova was 1.00${\pm}$0.00, 2.33${\pm}$0.52 and 2.20${\pm}$0.45, respectively, after receiving 100, 200 and 300 mg CAPE. The number of ovulated ova was 2.00${\pm}$0.00, 2.86${\pm}$0.69 and 3.00${\pm}$1.22, respectively, after receiving 10, 15 and 20 mg ACAPE. The fertilized and cultured ova were able to develop into embryos up to the 32 cell stage. The present experiments demonstrated that multiple ovulations can be induced by CAPE and ACAPE successfully, and the ova resulted from the treatment retained the capability for further fertilization and embryonic development. These data provide new information to support the establishment of an in vitro culture system for future avian transgenesis studies.

• Development and Reproduction
• /
• v.24 no.4
• /
• pp.297-306
• /
• 2020

Repetitive changes in the intracellular calcium concentration ([Ca²⁺]i) triggers egg activation, including cortical granule exocytosis, resumption of second meiosis, block to polyspermy, and initiating embryonic development. [Ca²⁺]i oscillations that continue for several hours, are required for the early events of egg activation and possibly connected to further development to the blastocyst stage. The sources of Ca²⁺ ion elevation during [Ca²⁺]i oscillations are Ca²⁺ release from endoplasmic reticulum through inositol 1,4,5 tri-phosphate receptor and Ca²⁺ ion influx through Ca²⁺ channel on the plasma membrane. Ca²⁺ channels have been characterized into voltage-dependent Ca²⁺ channels (VDCCs), ligand-gated Ca²⁺ channel, and leak-channel. VDCCs expressed on muscle cell or neuron is specified into L, T, N, P, Q, and R type VDCs by their activation threshold or their sensitivity to peptide toxins isolated from cone snails and spiders. The present study was aimed to investigate the localization pattern of N and P/Q type voltage-dependent calcium channels in mouse eggs and the role in fertilization. [Ca²⁺]i oscillation was observed in a Ca²⁺ contained medium with sperm factor or adenophostin A injection but disappeared in Ca²⁺ free medium. Ca²⁺ influx was decreased by Lat A. N-VDCC specific inhibitor, ω-Conotoxin CVIIA induced abnormal [Ca²⁺]i oscillation profiles in SrCl₂ treatment. N or P/Q type VDC were distributed on the plasma membrane in cortical cluster form, not in the cytoplasm. Ca²⁺ influx is essential for [Ca²⁺]i oscillation during mammalian fertilization. This Ca²⁺ influx might be controlled through the N or P/Q type VDCCs. Abnormal VDCCs expression of eggs could be tested in fertilization failure or low fertilization eggs in subfertility women.

• Proceedings of the Korean Society of Veterinary Clinics Conference
• /
• 2008.05a
• /
• pp.105-105
• /
• 2008

• Clinical and Experimental Reproductive Medicine
• /
• v.19 no.2
• /
• pp.163-168
• /
• 1992

This study was carried out to set up the ovum bank for ovum donation and to determine the best freezing method for human immature oocytes. Human immature follicular oocytes were cryopreserved by slow freezing and rapid thawing method. Immature follicular oocytes were treated by propanediol(PROH) solution by 2 and 4 step method in protocols A & B, respectively. In protocol C, immature oocytes were exposed to sucrose prior to treatment of PROH by 4 step method. We compared survival rate, maturation rate, and fertilization rate of immature oocytes among three protocols. Results were as follows. 1. Oocytes treated by the protocol C showed the highest survival rate( 70.3 %) and maturation rate(34.6%) after thawing. 2. Survival rate of oocytes treated by the protocol C was significantly higher than that of the protocol B after thawing(p<0.05). In conclusion, treatment of oocytes with sucrose prior to expose PROH was the best freezing method. Sucrose may have reduced the toxic effect of cryoprotectant to oocytes. We failed to induce fertilization of oocytes, which were treated by any protocols, by conventional insemination method, but obtained 28.8% fertilization rate by using partial zona dissection(PZD) method. This result suggests that micromanipulation(PZD) of the thawed oocytes before insemination will improve the fertilization rate.

• Magazine of the Korean Society of Agricultural Engineers
• /
• v.41 no.2
• /
• pp.44-54
• /
• 1999

Agricultural water quality standards were reviewed through rice culture using treated sewage irrigation . The seqage from school building of Konkuk University was treated by a constructed wetland system, and theeffluent of the systeml was irrigated for rice culutre after nutrient concentration adjusted by dilution. Average concentration of COD, SS, T-N and T-P in irrigated water was 22.3mg/$\ell$, 6.5mg/$\ell$, 25.8 mg/$\ell$and 2.2mg/$\ell$, respectively. Treatment include irrigation of adjusted effluent with conventional fertilization (TWCF), adjusted effluent with no fertilization (TWNF). and effluent of the wetland system as it was with no fertilization (SWNF). These treatment plots were compared with control plot irrigated by tap water with conventional fertilization (CONTROL). Other environmentals for rice culture were identical for all the plots. Among them, TWCF showed the best growth rate and the highest yield, and constituents in the harvested rice showed not much difference among them. Which implies that irrigation with relatively high nutrient concentration compared to the current water quality standards may cause no adverse effect on rice culture and could be even beneficial . Although T-N for this study was 25 times greater than the current standards, rice culture wasnot adversely affected by irrigatino water quality and even beeter results were observed than the CONTROL. It could be mistakenly that clean irrigation water produces better agricultural product, however, it is not necessarily true. Irrigation water with moderate nutrient concentration can enhance the plant growth, and better result might be expected. Therefore, peer review and modification if necessary are needed to the current agricultural water quality standards, especially for the nutrient components.

• Journal of Aquaculture
• /
• v.8 no.3
• /
• pp.159-170
• /
• 1995

Triploid abalone, Haliotis discus hannai, was induced by the cold $(0^{\circ}C\;and\;3^{\circ}C)$ or the heat $(35^{\circ}C\;and\;40^{\circ}C)$ shock procedure with fertilized eggs, 12 min. or 32 min. post fertilization with the various time intervals of shock duration. Fertilization rate of each experimental group was not significantly different from that of corresponding diploid control (P>0.05). However hatching rates and normality rates of triploid larvae were significantly different from those of corresponding diploid control (P <0.05). In heat shock groups at $40^{\circ}C$, fertilization rate of eggs was extremely low $(0\~2.7\%)$ and hatched larvae were not detected in these treatment groups. Incidence of triploidy was confirmed by chromosome count and the highest rates of triploid $(84.0\%)$ revealed in cold $(3^{\circ}C)$ shock with 15 min. treatment duration 12 min. after fertilization. The number of diploid chromosome was 2n: 36, and that of tiploid was 3n=54.

• Clinical and Experimental Reproductive Medicine
• /
• v.27 no.2
• /
• pp.201-209
• /
• 2000

Objective: This study has been carried out to evaluation the effect of fertilization promoting peptide (FPP) on the kinematic parameters, capacitation and acrosome reaction of the frozen-thawed human spermatozoa. Methods: After FPP treatment, we examined kinematic parameters, capacitation and acrosome reaction, using the methods of computer-aided sperm analysis (CASA) and chlortetracycline (CTC) fluorescence analysis. Results: We have obtained the evidence that FPP can promote the capacitation and inhibit the spontaneous acrosome reaction of frozen-thawed human spermatozoa in vitro. FPP ($25{\sim}100$ nM) induced a significant increase in the proportion of B-pattem capacitated spermatozoa, and a significant decrease in the proportion of F-pattem uncapacitated ones without significant stimulation of acrosomal exocytosis. In the kinematic parameters treatment, FPP treated groups maint3ined higher LIN, BCF and STR than those of control. The VAP, VSL, VCL and ALH were not different. Therefore it is suggested that FPP in human seminal plasma may play a positive role in promoting human sperm function.

• Korean Journal of Soil Science and Fertilizer
• /
• v.46 no.3
• /
• pp.216-222
• /
• 2013

Fertilizer management has the potential to promote the storage of carbon and nitrogen in agricultural soils and thus may contribute to crop sustainability and mitigation of global warming. In this study, the effects of fertilizer practices [no fertilizer (Control), chemical fertilizer (NPK), Compost, and chemical fertilizer plus compost] on soil total carbon (TC) and total nitrogen (TN) contents in inner soil profiles of paddy soil at 0-60 cm depth were examined by using long-term field experimental site at $42^{nd}$ years after installation. TC and TN concentrations of the treatments which included N input (NPK, Compost, NPK+Compost) in plow layer (0-15 cm) ranged from 19.0 to 26.4 g $kg^{-1}$ and 2.15 to 2.53 g $kg^{-1}$, respectively. Compared with control treatment, SOC (soil organic C) and TN concentrations were increased by 24.1 and 31.0%, 57.6 and 49.7%, and 72.2 and 54.5% for NPK, Compost, and NPK+Compost, respectively. However, long term fertilization significantly influenced TC concentration and pools to 30 cm depth. TC and TN pools for NPK, Compost, NPK+Compost in 0-30 cm depth ranged from 44.8 to 56.8 Mg $ha^{-1}$ and 5.78 to 6.49 Mg $ha^{-1}$, respectively. TC and TN pools were greater by 10.5 and 21.4%, 30.3 and 29.6%, and 39.9 and 36.3% in N input treatments (NPK, Compost, NPK+Compost) than in control treatment. These resulted from the formation and stability of aggregate in paddy soil with continuous mono rice cultivation. Therefore, fertilization practice could contribute to the storage of C and N in paddy soil, especially, organic amendments with chemical fertilizers may be alternative practices to sequester carbon and nitrogen in agricultural soil.

• Asian-Australasian Journal of Animal Sciences
• /
• v.24 no.12
• /
• pp.1706-1710
• /
• 2011

An experiment was conducted to investigate the effects of calcium (Ca) fertilization on oxalate content in napiergrass (Pennisetum purpureum) and on some blood parameters of sheep. Re-growth napiergrass was grown with or without Ca fertilizer and offered to sheep. Eight sheep, divided into two groups of 4 animals each were used. Calcium-fertilized napiergrass was offered daily to the animals as experimental treatment, whereas without Ca-fertilized napiergrass was given to the animals as control. Results showed that there was a trend to lower soluble and total oxalate concentrations in Ca-fertilized grass than control. The Ca-fertilized grass contained (p<0.05) lower concentrations of K and Mg than control, though Ca fertilization had no affect (p>0.05) on Ca and Na concentrations in plants. Feeding of Ca-fertilized grass had no affect on the feed consumption, blood Mg level and daily gain of sheep. However, sheep fed control grass had lower (p<0.05) blood Ca level than sheep fed Ca-fertilized grass. Our findings suggest that Ca fertilization might minimize the negative effects of oxalate.