This study is investigated the growth characteristics(number of available stipe, pileus diameter, pileus thickness, stipe length, stipe thickness, object weight, comparison yield ability of 1 cycle) and storage characteristics of ‘Sanjo 701’ (S7) cultivars according to relative humidity. The S7 growth characteristics were investigated by quantifying the growth and the characteristics according to the relative humidity, The storage stability was investigated every 5 days and freshness was measured by ‘Minamide Method’. S7 pileus diameter is The higher the relative humidity was confirmed becomes larger and the more increased relative humidity also increase the comparison yield ability of 1 cycle. However, pileus diameter or stipe length This could not see the big difference in the three treatment groups, the plieus thickness was no significant difference in the treatment of 80% and 95%. The fresh weight of S7 decreased significantly at 80% and 95% relative humidity after 10 days of storage, but decreased continuity in 65% humiditiry. The elongation percentage of S7 pileus was observe in 95% relative humidity, values of L, a and b (SCI), showed the highest L value in 65% and the a value in 95%. b values were similar in 65%, 80% and 95% treatments. On the ‘Minamide Method’ measure freshness was changed from the 10th day of storage at relative humidity of 95%, but humidity of 65% and 80% treatments, it changed after 15 days of storage. In this study growth characteristics and yield were increased at higher relative humidity, but storage stability was decreased. Therefore, it is deemed necessary to change the relative humidity to produce high quality mushrooms.

This research was relative humidity (65, 75, 85 and 95%) growth characteristics of the oak mushroom (Lentinula edodes) 'Nongjingo’. At the relative humidity was 65% in growth chamber. On this condition the moisture content of medium surface was 60.5%. That was lower than other treatments. That condition makes Pileus was 77.1% and stipe was 65%. Among the growth characteristics according to the relative humidity, the pileus diameter at 75% relative humidity were 45.5mm that largest than other treatment and pileus thickness at 75% relative humidity were largest than other treatment. The stipe thickness at 65% relative humidity were 14.8mm that lagest than other treatment. The number of available stipes at 95% relative humidity were 13.3peace/2kg that largest than other treatment. The comparison yield ability of 1 cycle was 104.2g that result in largest than other treatment at 95% relative humidity. This result was associated to the relative humidity of fruiting bodies, the lower relative humidity result in the lower of yield. However, the lower relative humidity result in the higher number of whago, Especially, at the treatment of 65% relative humidity, all the fruit bodies were produced as Whago (dry). Therefore, the higher relative humidity influenced in the higher yield but produced water mushroom. That result was thought that ‘Nongjingo’ would be possible to produce high quality mushroom by adjusting the relative humidity appropriately.

Recently, we published a microinjection method for generating transgenic cattle using the DNA transposon system and their analysis by next-generation sequencing (Yum et al. Sci Rep. 2016 Jun 21;6:27185). In that study, we generated transgenic cattle using two different types of DNA transposon system, sleeping beauty (SB) and piggybac (PB), carrying Yellow fluorescent protein with SB (SB-YFP, female) and green fluorescent protein with PB (PB-GFP, male) under the control of the ubiquitous CAG promoter, respectively. The female and male founder cattle have been grown up to date (the female age: 40 months old, the male age: 33 months old) without any health issues. In genomic instability and blood analysis, there was no significant differences between wild type and founder cattle. In the present study, we confirmed germ-line transmission of the transposon-mediated transgene integrations and ubiquitous and persistent expression of transgene in second generation of offspring (F1). The F1 was born without any assistance and expressed GFP in the eyes without UV light. The ubiquitous expression of GFP was detected in skin fibroblast from the ear tissue and confirmed by genomic DNA PCR, which suggest that the transgene from the PB-GFP was successfully transmitted. Unfortunately, no transgene from SB-YFP were identified. To confirm the transgene integration site, the genomic DNA from blood was extracted and performed next-generation sequencing (NGS). The GFP gene was integrated in chromosome 4 (two copies), and 6. As results, a total of two copies of paternal transgene transmitted into the F1. All the integrated position was not related with coding region and there was no significant difference in genomic variants between transgenic and non-transgenic cattle. To our knowledge, this is the first report of germ-line transmission through non-viral transgenic founder cattle. Those transgenic cattle will be valuable resource to many fields of biomedical research and agricultural science.

CRISPR/Cas9-induced knock-out/-in can be occurred at specific locus in the genome by non-homologous end joining (NHEJ) or homology directed repair (HDR). Here, we demonstrate the targeted insertion into the specific loci of embryo fertilized by semen from transgenic cattle via CRISPR/Cas9 system. Recently, we published on the efficient generation of transgenic cattle using the DNA transposon system (Yum et al. Sci Rep. 2016 Jun 21;6:27185). In the study, eight transgenic cattle were born following transposon-mediated gene delivery system (Sleeping Beauty and Piggybac transposon system) via microinjection. In the analysis of their genome stability using next-generation sequencing, there was no significant difference in the number of genetic variants between transgenic and non-transgenic cattle. All the transgenic cattle have grown up to date (the oldest age: 33 months old, the youngest age: 15 months old) without any health issue. One of transgenic male cattle expressing GFP reached puberty and semen was collected. Over 200 frozen semen straws were produced and some were used for in vitro fertilization (IVF). On seven days after IVF, expression of GFP was observed at blastocyst stage and was seen in 80% of the embryos. Another application is to edit the GFP locus of the transgenic cattle because long-term and ubiquitous expression of transgene didn’t affect their health. In one cell stage embryos produced using GFP frozen-thawed semen, microinjection of sgRNA for GFP, Cas9, together with donor DNA that included RFP and homology arms to link the double-strand break of sgRNA target site into fertilized eggs resulted in expression of RFP. This indicated that the GFP locus of transgenic cattle shows potential candidates for stable insertion of the functional transgene. Knock-out/-in for editing GFP locus using CRISPR-Cas9 might be a valuable approach for the next generation of transgenic models by microinjection. In conclusion, we demonstrated P-112 that transgenic cattle via transposon system are healthy to date and germ-line competence was confirmed. The GFP locus will be used as the potential target site for future gene engineering via genome-editing technology. Finally, all those animals could be a valuable agricultural and veterinary science resource for studying the effects of gene manipulation on biomedical research and medicine. This work was supported by BK21 PLUS Program for Creative Veterinary Science and Seoul Milk Coop (SNU 550-20160004).

The CRISPR/Cas9 system is proved to be a powerful tool for knock-out and knock-in in various species. By introducing genetic materials of two components (Cas9 and small guide (sg) RNA) into cells or pronuclear of the fertilized embryo, gene editing occurs. Some studies reported that efficiency of gene editing would be increased as Cas9 was integrated into cells or animals since Cas9 is indispensable in the CRISPR/Cas9 system. Accordingly, the production of Cas9 expressing cattle may provide the broadly used gene editing platform in cattle. For this study, Cas9 and RFP genes were cloned into PiggyBac (PB) transposon system. PB-Cas9-RFP and transposase were microinjected into 1436 in vitro fertilized embryos and 241 blastocysts were formed. Blastocysts with RFP expression accounting for 14.1% of total formed blastocysts were selected and transferred into 5 recipient cow. After gestation periods, four transgenic cattle were delivered without any veterinary assistance. From a transgenic cattle, ear skin tissue was collected for primary culture. On those primary cells, sgRNAs in DNA form for various genes such as PRNP, RB1 and BLG were transfected as 2ug of sgRNA per 5x105 cells using Nucleo factor system (Neon®, invitrogen, program#16). As expected, every group of each sgRNA delivered was confirmed to be mutated by T7E1assay. Those data demonstrated that for the first time, transgenic cattle with Cas9 expression were born, grown up to date and will be avaluable resource for genome-editing in cattle. This work was supported by BK21PLUS Program for Creative Veterinary Science and Seoul Milk Coop (SNU550-20160004).

This study investigated the antibacterial effects of Galla rhois extract (GRE) against Campylobacter jejuni and Campylobacter coli. The minimum inhibitory concentrations (MICs) of GRE against C. jejuni and C. coli were 0.28 and 0.55 mg/mL, respectively, and the corresponding minimum bactericidal concentrations (MBCs) were 4.4 and 5.5 mg/mL. C. jejuni treated with the MIC, MBC or 2×MBC of GRE showed significant inhibition of growth compared with that of the control group during the incubation period, and no viable bacteria were detected at 24 h after incubation. C. coli treated with MIC, MBC or 2×MBC of GRE also showed inhibition of growth compared with that of the control group during the incubation period, and in the C. coli cultures treated with MBC and 2×MBC of GRE, no viable bacteria were detected at 24 h after incubation. In conclusion, GRE is a candidate antibacterial agent against C. jejuni and C. coli, and may have applications for the control of Campylobacter infection in poultry.

농촌체험의 관심과 체험이 증가되는 상황에서 농촌체험의 지속성과 경쟁력의 확보 차원으로 서비스품질이 중요해지고 있다. 이 연구는 농촌체험을 경험한 서울지역 소비자를 대상으로 서비스품질과 고객만족, 고객만족과 추천의도 간의 관계를 분석하였다. 설문분석은 총 234부이며, 빈도분석, 신뢰도분석, 확인적 요인분석, 가설검정을 하였다. 분석결과는 다음과 같다. 첫째, 신뢰도의 크론바흐 알파값은 0.6 이상, 타당도의 요인적재량은 0.7 이상, 평균분산추출은 0.5 이상이 었다. 둘째, 확인적 요인분석의 모형적합도는, CMIN/DF, GFI, TLI, CFI, RMSEA이 전반적으로 적합하였다. 셋째, 가설검정의 결과, H1의 H1-2(유형성)와 H1-3(응답성)을 제외한 H1-1(신뢰성), H1-4(확신성), H1-5(공감성)는 고객만족 에 긍정적인 영향을 주었고, H2는 고객만족이 추천의도에 긍정적인 영향을 주었다. 따라서 농촌체험의 서비스를 이용자와 제공자 간의 상호작용으로 볼 때, 서비스품질에 대한 지속적인 개선이 필요할 것이다.

Total phenol contents, total flavonoid contents and antioxidant activity of 70% methanol, 70% ethanol and chloroform-methanol (CM, 2:1, v/v) extracts from onion (Allium cepa L.) peels were studied. The IC50 values of DPPH (2,2-diphenyl-1-picrylhydrazyl) radical and ABTS [2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt] radical scavenging activity in 70% ethanol extract were remained to be lowest followed by 70% methanol extract and CM extract. And the total phenol content (113.56±0.86 mg CAE/g), total flavonoid content (49.63 mg QE/g) and ferric reducing antioxidant power value were also found to be the highest. In contrast, 70% methanol extract possessed the strongest antioxidant activity by β-carotene bleaching assay. CM extract displayed the lowest antioxidant activity compared with other extracts. Onion peels exhibited strong antioxidant activity and abundant phytochemicals, which could be used in a various food products to add phytochemicals and promote good health.

목 적: 본 연구는 환경 강화를 다룬 성인 쥐 모델에서의 증거들을 비판적으로 검토하고, 이를 다른 동물 과 인간에게 적용 가능한지에 대해 논의하고자 하였다. 방 법: 연구진은 환경 강화와 관련된 성인 쥐 연구들을 2014년 7월까지 서지 데이터베이스(PubMed, Medline, EMBASE, Web of Science and CINAHL)에서 검색하였고, 포함된 연구들의 참고 문헌에서 본 연 구와 관련된 선행 연구들을 또한 포함시켰다. 방법론적 평가는 본 연구의 세 명의 저자가 독립적으로 구체화 된 요소들을 가지고 시행하였다. 결 과: 포함 및 불포함 기준을 적용하여 최종적으로 15개의 연구가 채택되었다. 모든 포함된 연구들은 일관적으로 시각겉질의 가소성을 증진시킴으로써 성인 쥐에서의 시각 향상의 증거를 보여주었고, 이러한 가 소성을 증진시키는 요소들로는 해부학적 변화에 의한 가소성 재개, 시력 향상, 눈 우세성(ocular dominance)의 변화, 행동적 시력 향상들이 있었다. 이러한 요소들에도 불구하고, 포함된 연구들에서 방법론적 약점들이 내재하고 있다는 것을 발견하였다. 결 론: 본 체계적 문헌 고찰의 결과들을 통해 약시를 가진 성인 쥐의 시각겉질이 결정적 시기가 지났음 에도 불구하고 환경 강화에 대응하여 반응하는 가소성을 가지고 있다는 것을 제안한다. 뿐만 아니라 약시를 가진 성인 시각겉질에 환경 강화를 적용하여 시각계의 재활을 도울 수 있을 것으로 간주되지만, 이를 위해서는 인간을 포함한 다른 동물들 간의 해부학적 및 생활방식의 차이들을 극복해야만 가능하다고 본다.

To date, most serodiagnostic methods for brucellosis screening are based on antibodies against lipopolysaccharides of Brucella spp. However, this approach has the drawback of yielding false-positive results due to cross-reactivity with lipopolysaccharides of other related pathogens, especially Yersinia enterocolitica O:9. In this study, Brucella abortus AspC was cloned and expressed by PCR amplification into a pCold TF expression system to obtain recombinant AspC (rAspC). The immunogenicity of rAspC was confirmed by western blotting of Brucella-positive bovine serum. rAspC-based ELISA was performed to determine whether rAspC could be used in the serodiagnosis of bovine brucellosis. rAspC reacted strongly with anti-Brucella antibodies in positive sera in the tube agglutination test (TAT), but did not show strong reaction with most negative samples. In particular, the average OD492 value at the highest TAT titer showed a 1.4-fold increase with respect to the cutoff value. The accuracy, specificity, and sensitivity of rAspC were 71.88%, 78.33%, and 68%, respectively. These findings suggest that rAspC might be valuable for the serological diagnosis of bovine brucellosis.

The aim of this study was to investigate the antioxidant activity of thermal treatment yam (Dioscorea batatas DECNE.) in Korea. Thermal treatment yam was extracted by different solvents including 70% methanol, 70% ethanol and chloroform–methanol mixture (CM, 2:1, v/v). Then color property, total phenol content and antioxidant activity were analyzed. Yam possessed high L* value and H° value, which were 54.92±2.18 and 73.20±0.77, respectively. Thermal treatment yam exhibited great antioxidant activity evaluated by ABTS [2,2′ -azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt] radical scavenging activity, DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, reducing power, and ferric reducing antioxidant power. Total phenol contents of various extracts from thermal treatment yam increased in the following order: 70% methanol extract (63.53±0.33 mg CAE/g), 70% ethanol extract (69.47±1.00 mg CAE/g) and CM extract (97.49±0.66 mg CAE/g), respectively. The same trend was also could be found in antioxidant activity assays except for reducing power assay. These results implied that these extracts from thermal treatment yam might be useful to take a good part in prevention of human diseases and aging.

In this study, the thermal treatment yam (Dioscorea batatas DECNE.) was analyzed for its proximate and nutritional compositions including mineral, vitamin, amino acids and fatty acids. Thermal treatment yam is mainly consisted of carbohydrate (70.3%) with small amounts of crude protein (15.8%), crude ash (5.1%) and crude fat (1.6%). It was found to be a good source of essential minerals such as K (1100.2 mg/100 g), Mg (99.4 mg/100 g), Ca (54.8 mg/100 g) and Na (44.9 mg/100 g) but Zn (4.8 mg/100 g) and Fe (0.2 mg/100 g) content was low. Little amounts of vitamin E (8.3 mg/100 g), vitamin C (3.5 mg/100 g) and vitamin B1 (2.1 mg/100 g) could be found. The amino acid analysis revealed that the yam was superior with respect to serine (1454.2 mg%), lysine (684.6 mg%) and histidine (684.6 mg%). Essential amino acids were calculated to be 2849.3 mg%. The amino acid profiles showed that thermal treatment yam is limiting in isoleucine and phenylaline. Linoleic acid was the most predominant fatty acids with the value of 47.3% followed by palmitic acid (24.7%) and oleic acid (12.7%). And the unsaturated fatty acids including oleic acid and linoleic acid were present in big quantities in thermal treatment yam.

Brucellosis is an important and re-emerging zoonotic disease worldwide. The prevention of human infection is achieved predominantly through the control of brucellosis in agricultural animals, which in turn depends on accurate diagnosis and vaccination. However, conventional serological diagnosis of brucellosis has several limitations, and currently available vaccines for animals have several drawbacks, including the ability to cause infection in humans. Phosphoglycerate kinase (Pgk) is one of the specific proteins reactive with mouse sera in the early stage of Brucella infection, and deletion of the pgk gene in B. abortus strain 2308 resulted in extreme attenuation of this strain in vitro and in vivo. Furthermore, the B. abortus pgk mutant has been used as a live vaccine, and in challenge experiments, it induced protection that was superior to that conferred by commercial strains. In this study, the pgk gene from Brucella abortus 544 was successfully amplified and cloned into a maltose binding protein fusion protein expression vector (pMAL). The recombinant protein was expressed in Escherichia coli DH5α and purified. The immunogenicity of purified recombinant B. abortus 544 Pgk (rPgk) was evaluated by western blot analysis using Brucella-positive mouse sera. rPgk could be used as an antigenic component for future serological tests and potential vaccine development.

본 연구는 오배자 에탄올 추출물 (GRE), 염소산나트륨 (SC) 그리고 오배자 에탄올 추출물과 염소산나트륨 합제(GS)의 B. abortus에 대한 항균효과를 확인하기 위해 수행 되었다. GRE, SC 그리고 GS를 B. abortus에 처리하여 배양한 후, B. abortus의 생존수를 확인하였으며, 마우스 탐식세포 내 감염된 B. abortus의 증식 억제효과를 경시별 (2, 24, 48시간)로 조사하였다. GRE, SC 그리고 GS는 각각 400 μg/mL 이하, 15 mM 그리고 0.6GS (GS 1, GRE 1,000 μg/mL + SC 30 mM) 이하의 농도에서 세포독성을 나타나지 않았다. 모든 처리구에서 B. abortus의 생존율은 용량- 의존적으로 현저하게 감소하는 결과를 나타내었다. 또한, GRE (400 μg/mL), SC (15 mM) 그리고 0.5GS (GRE 500 μg/mL + SC 15 mM)를 처리한 세포에서 배양 48시간 후에, B. abortus의 증식이 통계적으로 유의성 있게 감소하였으며 (GRE, p < 0.01; SC and 0.5GS, p < 0.001), 특히, GS를 처리한 경우, B. abortus의 세포내 증식이 GRE와 SC의 상승작용에 의한 강력한 항균효과를 나타내었다. 결론적으로, GS는 B. abortus에 대한 항균물질로서 유용할 뿐만 아니라, 식육과 우유 위생 분야에 적용할 수 있을 것으로 생각된다.

Brucellosis is a notorious zoonotic disease with global implications. Efforts to control the spread of the disease have been restricted to the agricultural livestock. Increasing incidences of accidental human infection have motivated researches to start working on alternative vaccines. At present, live attenuated vaccines are the only accepted type of vaccines used in developed countries for the prevention of brucellosis. Although serodiagnosis is occasionally unreliable, some countries have already claimed to have eradicated the disease, based on this testing. Live attenuated vaccines are not suitable for use in pregnant and immune-depressed animals. Moreover, these vaccines are not tolerated in humans. Therefore, many researches have been striving to discover alternative methods of vaccination. Most research has focused on the generation of subcellular, subunit, and DNA vaccines that are as efficient as the live attenuated vaccines. At present, none of the available vaccines has been able to replace the live attenuated vaccines. Therefore, additional research is necessary in order to discover a new brucellosis vaccine that is suitable for human use.