Canine parvovirus-2 (CPV-2) has been reported worldwide as a major pathogen associated with acute hemorrhagic enteritis. The disease is a major infectious cause of death, particularly in young dogs. The earliest type of CPV-2 was replaced with three main subspecies, CPV-2a, CPV-2b, and CPV-2c, within a few years. Vaccination is carried out regularly, but the emergence of antigenic variants and the influence of maternal antibodies have limited the efficacy of commercial vaccines. New vaccines, such as the subunit vaccine, have been developed for alternative, safe, and effective vaccination. The baculovirus expression vector system (BEVS) is an excellent eukaryotic expression system with a high-level expression of foreign proteins and the ability of post-translational modification. Therefore, it is used widely to produce recombinant protein and subunit vaccines. In this study, the VP2 protein of CPV-2b cloned in the gateway vector system was generated using a baculovirus expression system in Spodoptera frugiperda (SF9) insect cells. Hemagglutination assay (HA) titers (24) were obtained, and the expression was detected in 6-His tagged VP2 and monoclonal antibody (mAb) against CPV-2 by western blotting. The VP2 protein of CPV-2b expressed in this study may provide a basis for a clinical diagnosis and vaccination applications for CPV-2.

Pleurotus ostreatus is a globally cultivated mushroom crop. Cap color is a quality factor in P. ostreatus. However, cap color can spontaneously mutate, degrading the quality of the mushroom on the market. Early detection and removal of mutant strains is the best way to maintain the commercial value of the crop. To detect the cap color mutant Gonji7ho, molecular markers were developed based on insertion/deletions (InDels) derived from the comparison of mitogenomes of Gonji7ho and Gonji7hoM mushrooms. Sequencing, assembly, and comparative analysis of the two mitogenomes revealed genome sizes of 73,212 bp and 72,576 bp with 61 and 57 genes or open reading frames (ORFs) in P. ostreatus Gonji7ho and Gonji7hoM, respectively. Fourteen core protein-encoding genes, two rRNA, and 24 tRNA with some OFRs were predicted. Of the 61 genes or OFRs in the wild type, dpo, rpo, and two orf139 were missing (or remnant) in the mutant strain. Molecular markers were developed based on the sequence variations (InDels) between the two mitogenomes. Six polymorphic molecular markers could detect the mutated mitochondria by PCR. These results provide basic knowledge of the mitogenomes of wild-type and mutant P. ostreatus, and can be applied to discriminate mutated mitochondria.

본 연구는 특수용도식품에 함유되어 있는 비타민B12와 비오틴에 대하여 식품의 복잡한 매질 특성에 특이성을 확보할 수 있도록 현행 식품공전 시험법을 개선하고자 하였다. 식품공전에서 비타민B12와 비오틴의 시험법에 항체반응을 통해서 분석물질과 특이적으로 결합하여 그 외의 성분들을 효과적으로 제거할 수 있는 면역친화성 컬럼을 도입하여 시험용액을 정제 및 농축함으로써 전처리 과정을 개선하였다. 식품 등 시험법 마련 표준절차에 관한 가이 드라인에 따라 밸리데이션(특이성, 직선성, 검출한계, 정량 한계, 정확도, 정밀도)을 수행하고 개선된 시험법을 이용하여 유통 중인 특수용도식품의 대상 품목에 대해 시험법 적용여부를 확인하였다. 본 연구사업의 수행으로 시험·검사의 정확성을 증대시켜 특수용도식품 품질관리에 대한 신뢰성을 더 높일 것으로 본다.

Oyster mushroom is one of mushrooms that are cultivated and consumed a lot in Korea. P. ostreatus 'ASI 2504(Suhan)' is a preferable cultivar to mushroom farmers because it has a dark pileus and a thick stipe. But it is very sensitive to environmental conditions, so farmers demand an alternative cultivar of ‘Suhan’ continuously. To develop a new cultivar, parental strains ‘ASI 2504(Suhan)’ and ‘ASI 0665(Heuktari)’ were selected from P. ostreatus’s various collected strains according to morphological characteristics. P. ostreatus ‘Soltari’ was developed by the method of Di-Mono crossing between dikaryotic strain ‘Suhan’ and monokaryotic strain derived from ‘Heuktari’. Analysis of the mitochondrial genetic characteristics was performed for primary selection in 100 crossed strains. The mitochondrial DNA profile of ‘Soltari’ was same as that of ‘Heuktari’, when mitochondrial DNA primer MtPo1 was used. And a nuclear DNA profile of ‘Soltari’ was similar as those of the parental strains, ‘Suhan’ and ‘Heuktari’, when RAPD(Random Amplified Polymorphic DNA) primer URP 1, 3 were used. The optimum temperature for mycelial growth was 30°C for ‘Soltari’. ‘Soltari’ was appropriate for middle high temperature to grow, especially 13~18°C. Fruiting body production per bottle (1,100mL) was about 158.6g. When compared to the control strain ‘Suhan’, the stipe’s length and thickness of ‘Soltari’ were similar to those of ’Suhan’. But the pileus diameter of ‘Soltari’ was a little shorter than that of ‘Suhan’, the former was 42.72mm, while the latter was 51.33mm. And the pileus thickness of ‘Soltari’ and ‘Suhan’ were 18.18mm and 25.46mm, respectively. ‘Soltari’ was more resistant at high CO2 concentration than ‘Suhan’ and the color of pileus of ‘Soltari’ was kept dark gray at high temperature. Therefore, it is suggested that this new cultivar ‘Soltari’ be an alternative of ‘Suhan’ and contribute to energy saving effect in oyster mushroom farms.

Oyster mushroom is one of mushrooms that are cultivated and consumed a lot in Korea. P. ostreatus 'Suhan(ASI2504)' is a popluar cultivar because of high quality. But it is difficult for farmers to cultivate, so an alternative cultivar of ‘Suhan’ is demanded continuously. To develop a new variety, parental strains were selected using cultivation characteristics of Pleurotus ostreatus collected home and abroad. P. ostreatus 'Gosol' was developed by the method of Di-Mon crossing between dikaryotic strain P. ostreatus 'Mongdol(ASI0633)' and monokaryotic strain derived from P. ostreatus ‘Yasan(ASI0635)' in 2014. Analysis of the genetic characteristics of the new cultivar 'Gosol' showed a different DNA profile as that of the control strain, 'Suhan’ but a similar DNA profile as those of the parental strains, ‘Yasan’ and　'Mongdol’, when RAPD(Random Amplified Polymorphic DNA) primer URP 3 was used. The optimum temperature for mycelial growth was 25°C for ‘Gosol’ and ‘Suhan’. 'Gosol' was appropriate for middle high temperature to grow, especially 13~18°C. Fruiting body production per bottle was about 124.2g. When compared to the control strain 'Suhan’, the stipe was longer and the individual weight was heavier. But the stipe and the pileus were a little thinner than those of the control strain. ‘Gosol’ was more resistant at high CO2 concentration than the control strain. This new cultivar ‘Gosol’ of Pleurotus ostreatus was characterized dark bluish gray cultivar of oyster mushroom in the color of pileus and higer yield compared to those of other cultivar ‘Suhan’. Therefore, it is suggested that this new cultivar ‘Gosol’ be substitute for ‘Suhan’ in oyster mushroom’s farms.

The activation of glial cells in the spinal cord has been contribute to the initiation and maintenance of pain facilitation induced by peripheral inflammation and nerve injury. The present study investigated effects of botulinum toxin type A (BoNT-A), injected subcutaneously or intracisternally, on the expression of microglia and astrocytes in rats. Complete Freund’s Adjuvant (CFA)-induced inflammation was employed as an orofacial chronic inflammatory pain model. A subcutaneous injection of 40 μL CFA into the vibrissa pad was performed under 3% isoflurane anesthesia in SD rats. Immunohistochemical analysis for changes in Iba1 (a microglia marker) and GFAP (an astrocyte marker), were performed 5 days after CFA injection. Subcutaneous injection of CFA produced increases in Iba1 and GFAP expression, in the ipsilateral superficial lamia I and II in the medullary dorsal horn of rats. Subcutaneous treatment with BoNT-A attenuated the up-regulation of Iba1 and GFAP expressions induced by CFA injection. Moreover, intracisternal injection of BoNT-A also attenuated the up-regulated Iba1 and GFAP expressions. These results suggest that the anti-nociceptive action of BoNT-A is mediated by modulation activation of glial cells, including microglia and astrocyte.