• Fisheries and Aquatic Sciences
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• 제27권5호
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• pp.306-313
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• 2024

The genus Chirostoma is endemic from the Mesa Central of Mexico. It is conformed of 18 species and six subspecies. Five species are in some category of risk, because of this, Chirostoma jordani is an excellent model species to implement biotechnologies like gametes cryopreservation. Aim of present study was to evaluate fertilizing capacity of cryopreserved C. jordani sperm, as alternative to conservation and assisted reproduction in this specie and genus. Males and females were collected from wild Atlangatepec dam stock, Tlaxcala State, Mexico. Seminal quality was evaluated in fresh and cryopreserved semen with three cryoprotective agents (CPAs): 10% dimethyl sulfoxide (DMSO), 10% methanol (MeOH), 14% ethylene glycol (EG) and it was determined its post-thaw fertilizing capacity. Sperm motility percentage decreased during cryopreservation process (p < 0.05). There were not significant differences in post-thaw motility percentage between EG (53.5 ± 1.9%) and MeOH (53.3 ± 1.3%), but DMSO (50.3 ± 0.5%) was significantly different (p < 0.05). Results showed that 0.2 μL fresh semen were enough to fertilize 100% oocytes (n = 60). 10 μL DMSO and 5 μL MeOH and EG cryopreserved semen were necessary to fertilize oocytes 100% (n = 60) (p < 0.05). Cryopreservation and fertilization protocol for C. jordani sperm was efficient and it could be used for its assisted reproduction.

• Clinical and Experimental Reproductive Medicine
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• 제37권4호
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• pp.307-319
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• 2010

목적: 유리화 동결액의 조성조절과 냉각속도 증진을 통한 동결보호제의 농도를 낮추는 전략을 통해 세포에 미치는 독성을 감소시켜 유리화 동결 및 융해 후 생쥐 배아의 생존율 및 발생률을 증진시키고, 궁극적으로 배아의 유리화 동결법을 개선하고자 하였다. 연구방법: 생쥐 배아와 포배기를 그리드를 이용한 유리화 동결법을 이용하여 동결/융해하였다. 동결액 내 ethylene glycol와 dimethylsulphoxide (DMSO)의 농도와 당의 농도를 조절하여 생쥐 배아의 융해 후 생존율과 발생률을 관찰하였고, 냉각속도의 증가와 동결억제제의 농도와의 상관관계를 포배기의 융해 후 생존율과 발생률에 따라 비교하였다. 또한 융해 후 배아를 대리모에 이식하여 산자를 생산함으로 냉각속도의 효율성을 알아보았다. 결과: EG를 단독으로 사용한 동결보존액 보다는 DMSO와 혼합된 동결보존액의 사용이 보다 유리하다는 결과를 얻을 수 있었다. 슬러시 질소에 의한 냉각속도의 증가가 동결보존의 대상의 상해를 줄여 유리화 동결의 효율성을 증진시키는 것으로 생각된다. 결론: 혼합된 동결보호제를 사용하였을 때 생쥐 배아의 유리화 동결 후 생존과 발생률이 증진되었다. 슬러시 질소를 이용한 유리화 동결의 도입은 냉각속도의 증가를 통해 기존 유리화 동결방법의 효율을 증진시켜 생존율과 융해후 발생률, 임신율 증진에 기여하였다. 또한 냉각속도의 증진은 유리화 동결의 필수요건인 고농도의 동결억제제에 대한 노출을 감소시킬 수 있었다. 이러한 노력은 생식력의 보전을 위한 유리화 동결법의 효율 향상에 기여할 것이다.

• Journal of Microbiology and Biotechnology
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• 제28권5호
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• pp.718-731
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• 2018

The beneficial effects of lactic acid bacteria (LAB) have been intensively investigated in recent decades with special focus on modulation of the host intestinal microbiota. Numerous discoveries of effective probiotics are driven by a significantly increasing demand for dietary supplements. Consequently, technological advances in the large-scale production and lyophilization are needed by probiotic-related industries for producing probiotic LAB for commercial use. Our study had a dual objective, to determine the optimum growth medium composition and to investigate appropriate cryoprotective additives (CPAs) for Lactobacillus salivarius, and compare its responses with other Lactobacillus species. The one-factor-at-a-time method and central composite design were applied to determine the optimal medium composition for L. salivarius cultivation. The following composition of the medium was established (per liter): 21.64 g maltose, 85 g yeast extract, 1.21 ml Tween 80, 6 g sodium acetate, $0.2g\;MgSO_4{\cdot}7H_2O$, $0.02g\;MnSO_4{\cdot}H_2O$, $1g\;K_2HPO_4$, $1.5g\;KH_2PO_4$, $0.01g\;FeSO_4{\cdot}7H_2O$, and 1 g sodium citrate. A cryoprotective additive combination comprising 10% (w/v) skim milk and 10% (w/v) sucrose supplemented with 2.5% (w/v) sodium glutamate was selected for L. salivarius, and its effectiveness was confirmed using culture-independent methods in the freeze-dried cells of the Lactobacillus strains. In conclusion, the optimized medium enhanced the species-specific cultivation of L. salivarius. On the other hand, the cryoprotective effects of the selected CPA mixture may also be dependent on the bacterial strain. This study highlights the necessity for precise and advanced processing techniques for large-scale production of probiotics in the food and feed industries.

• 한국패류학회지
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• 제29권3호
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• pp.251-258
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• 2013

본 연구에서는 굴, Crassostrea gigas의 계획적인 인공종묘생산 및 우량형질의 정자를 보존하기 위하여 냉동보존 시 적정 결빙억제제 (cryprotective agent, CPA) 및 농도를 알아보고자, CPA 종류 및 농도별 독성 및 냉동보존 효과를 파악하고 해동 후 정자의 세포 손상을 조사하였다. CPA 종류 및 농도별 침지시간에 따른 굴 정자의 독성평가를 실시한 결과, 생존율 및 운동성은 DMSO가 가장 좋았으며, 다음으로 EG, glycerol, Methanol 순이었다. 희석액으로 여과해수를 사용하여 CPA 종류 및 농도별 냉동보존 결과, 15% DMSO로 냉동보존 한 정자의 생존율 및 운동성이 가장 높았다. 굴의 냉동/해동 정자를 주사전자현미경으로 관찰한 결과, DMSO, EG, methanol, glycerol 순으로 세포 손상이 적었으며, 농도는 15, 20, 10, 5% 순이었다. 이상의 결과를 종합한 결과, 굴 정자의 냉동보존 시 여과해수를 희석액으로 사용할 경우 적정 CPA는 DMSO이며, 농도는 15%였다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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• pp.62-62
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• 2019

Bacterial leaf blight(BLB), caused by X. oryzae pv. oryzae(Xoo), is one of the most destructive diseases of rice due to its high epidemic potential. Understanding BLB resistance at a genetic level is important to further improve the rice breeding that provides one of the best approaches to control BLB disease. In the present investigation, a collection of 192 accessions was used in the genome-wide association study (GWAS) for BLB resistance loci against four Korean races of Xoo that were represented by the prevailing BLB isolates under Xoo differential system. A total of 192 accessions of rice germplasm were selected on the basis of the bioassay using four isolated races of Xoo such as K1 and K2. The selected accessions was used to prepare 384-plex genotyping by sequencing (GBS) libraries and Illumina HiSeq 2000 pairedend read was used for GBS sequencing. GWAS was conducted using TASSEL 5.0. The TASSEL program uses a mixed linear model (MLM). The results of the bioassay using a selected set of 192 accessions showed that a large number of accessions (93.75%) were resistant to K1 race and K2 resistant germplasm proportion remained between 66.67. The genotypic data produced SNP matrix for a total of 293,379 SNPs. After imputation the missing data was removed, which exhibited 34,724 SNPs for association analysis. GWAS results showed strong signals of association at a threshold of [-log10(P-value)] more than 5 (K1 and K2) for nine of the 39 SNPs, which are plausible candidate loci of resistance genes. These SNP loci were positioned on rice chromosome 2, 9, and 11 for K1 and K2 races. The significant loci detected have also been illustrated and make the CPAS markers for NBS-LRR type disease resistance protein, SNARE domain containing protein, Histone deacetylase 19, NADP-dependent oxidoreductase, and other expressed and unknown proteins. Our results provide a better understanding of the distribution of genetic variation of BLB resistance to Korean pathogen races and breeding of resistant rice.

• 한국해양생명과학회지
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• 제8권2호
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• pp.115-120
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• 2023

본 연구는 동자개 정자의 동결보존을 위해 동결보존제의 최적 농도와 적정 희석액을 구명하여 정자를 최상의 상태로 보존하여 인공종자를 생산하는데 목적이 있다. 실험은 3종의 희석액(I: 300 mM glycose, II: Kurokura extender, III: Li extender), 4종의 동결보존제(dimethyl sulfoxide, ethylene glycol, methanol and glycerol)와 4개의 동결보존제 농도(5, 10, 15, 20%)를 조합하여 대하여 조사하였다. 동결보존한 정자를 해동한 후 정자의 생존율과 정자활성지수로 동결보존제의 효과를 평가하였다. 희석액 III(Li extender)과 10과 15%의 methanol을 조합했을 때 동자개 정자의 생존율과 정자활성지수는 각각 66.9 ± 8.7, 67.3 ± 13.1%과 2.6 ± 0.4, 2.6 ± 0.5로 다른 희석액과 동결보존제보다 높았다.