• Applied Microscopy
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• v.52
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• pp.6.1-6.5
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• 2022

We examined the morphology of the fertilized egg and the fine structure of fertilized egg envelopes of Poropanchax normani belonging to the family Poeciliidae, also known as Norman's lampeye using light and electron microscopes. The fertilized eggs with narrow perivitelline space were found to be spherical and demersal, additionally containing small oil droplets in the vitelline membrane. Further, a bundle of adhesive filaments was observed to be present on one side of the fertilized egg. These filaments possessed remarkably high elasticity and were approximately 1-3mm in length. The size of the fertilized egg was determined to be about 1.49 ± 0.07mm (n=30). The outer surface appeared smooth, and adhesive filaments originating at different location of the surface of the envelope were found to be distributed around the egg envelope and were joined together to form a single long bundle in scanning electron microscopic observation. A peak-like structure formed of several straight wrinkles was observed around the micropyle. However, the complete structure of the micropyle could not be studied due to the depth at which it was located. Additionally, the total thickness of the egg envelope was ascertained to be approximately12.5-14.5㎛. The egg envelope consisted of two distinct layers, an outer electron dense layer and an inner lamellar layer, further consisting of 10 sublayers of varying thicknesses. Collectively, it was observed that the morphological characteristics of the fertilized egg, fine structures surrounding the micropyle, outer surface, adhesive structure consisting adhesive filaments, and sections of fertilized egg envelope displayed species specificity.

• Applied Microscopy
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• v.51
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• pp.3.1-3.6
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• 2021

We examined the morphology of fertilized egg and ultrastructures of fertilized egg envelopes of dwarf rainbowfish (Melanotaenia praecox) belong to Melanotaeniidae using light and electron microscopes. The fertilized eggs were spherical with adhesive filament, transparent, demersal, and had a narrow perivitelline space and small oil droplets. The size of fertilized egg was 1.02 ± 0.18 mm (n = 30), and there were two kinds of adhesive filament on the fertilized eggs. The long and thick (diameter 12.22 ± 0.52 ㎛, n = 20) adhesive filaments were only at the area of animal pole, and short and thin (diameter 1.99 ± 0.23 ㎛, n = 20) adhesive filaments were around the long filaments. A micropyle was conical shaped with adhesive filament and located near the animal pole of egg. The outer surface of fertilized egg was rough side. Also, the total thickness of the fertilized egg envelope was about 7.46 ± 0.41 ㎛ (n = 20), the fertilized egg envelope consisted of two layers, an inner lamellae layer and an outer layer with high electron-density. And the inner layer was 8 layers. Collectively, these morphological characteristics and adhesive property of fertilized egg with adhesive filaments, and ultrastructures of micropyle, outer surface, and section of fertilized egg envelope are showed species specificity.

• Journal of Life Science
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• v.27 no.11
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• pp.1331-1339
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• 2017

DNA barcoding is the identification of a species based on the DNA sequence of a fragment of the cytochrome C oxidase subunit I (COI) gene in the mitochondrial genome. It is widely applied to assist with the sustainable development of fishery-product resources and the protection of fish biodiversity. This study attempted to verify horse-head fish (Branchiostegus japonicus) and fake horse-head fish (Branchiostegus albus) species, which are commonly consumed in Korea. For the validation of the two species, a real-time PCR method was developed based on the species' mitochondrial DNA genome. Inter-species variations in mitochondrial DNA were observed in a bioinformatics analysis of the mitochondrial genomic DNA sequences of the two species. Some highly conserved regions and a few other regions were identified in the mitochondrial COI of the species. In order to test whether variations in the sequences were definitive, primers that targeted the varied regions of COI were designed and applied to amplify the DNA using the real-time PCR system. Threshold-cycle (Ct) range results confirmed that the Ct ranges of the real-time PCR were identical to the expected species of origin. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct of B. japonicus DNA ($21.85{\pm}3.599$) and the average Ct of B. albus DNA ($33.49{\pm}1.183$) for confirming B. japonicus. The assays also showed statistically significant differences between the average Ct of B. albus DNA ($22.49{\pm}0.908$) and the average Ct of B. japonicus DNA ($33.93{\pm}0.479$) for confirming B. albus. The methodology was validated by using ten commercial samples. The genomic DNA-based molecular technique that used the real-time PCR was a reliable method for the taxonomic classification of animal tissues.

• The Korean Journal of Mycology
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• v.23 no.1 s.72
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• pp.92-104
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• 1995

Ten species of Cordyceps species were collected throughout Kangwon province including Chuncheon Dongsanmyun KNU forest experiment from June to September, 1993. Collected Cordyceps species were identified as Cordyceps militaris, C. roseostromata, C. kyushuensis, C. scarabaeicola, Phytocordyceps ninchukiospora, C. nutans, Paecilomyces tenuipes, C. sphecocephala, Hymenostilbe odonatae, Torrubiella sp.. C. militaris, type species of Cordyceps species, was mainly formed on pupae of Lepidoptera and found after the rainy season around July. Fruiting body of C. roseostromata was morphologically similar to those of C. militaris, but relatively small in size and they were also found on lawn or pupa of Lepidoptera. Fruiting body of C. scarabaeicola was found on adult Scarabaeidae specifically and collect fruiting bodies of C. kyushuensis were on larva of moth. C. nutans and C. sphecocephala had host specificity on Hemiptera and Hymenoptera, respectively. Each species formed elliptical fertile part attach to the slim and carneous stalk and they were collected the most in specimen number through whole season of the summer. Ascospore of Phytocordyceps ninchukiospora on seed was characterized by two viable, multiseptate, fusiform units linked end-to-end by a long, filiform connective. Paecilomyces tenuipes, imperfect stage of the genus Cordyceps is multi-infective fungi that attack all stages of all groups of insects. Hymenostilbe odonatae attacks only adult Odonata and Torrubiella sp. formed on spider was difficult to collect because it was found the back side of leaf. As results of cultural test PDA medium showed the best mycelial growth. In the experiment of effect of the acidity inside of the media, C. militaris was good on pH 5, C. nutans and Phytocordyceps ninchukiospora were good on pH 6 and Paecilomyces tenuipes was on pH 7 and C. scarabaeicola was on pH 9. All isolates tested showed the best mycelial growth at $20^{\circ}C$. Morphologically similar isolates were used to analyze protein banding pattern among and within species. As a result, C. militaris, C. roseostromata and C. kyushuensis were clustered into close species and C. scarabaeicola and Phytocordyceps ninchukiospora were relatively distant from those species.

• Toxicological Research
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• v.33 no.1
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• pp.43-48
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• 2017

With ultraviolet and visible light exposure, some pharmaceutical substances applied systemically or topically may cause phototoxic skin irritation. The major factor in phototoxicity is the generation of reactive oxygen species (ROS) such as singlet oxygen and superoxide anion that cause oxidative damage to DNA, lipids and proteins. Thus, measuring the generation of ROS can predict the phototoxic potential of a given substance indirectly. For this reason, a standard ROS assay (ROS assay) was developed and validated and provides an alternative method for phototoxicity evaluation. However, negative substances are over-predicted by the assay. Except for ultraviolet A (UVA), other UV ranges are not a major factor in causing phototoxicity and may lead to incorrect labeling of some non-phototoxic substances as being phototoxic in the ROS assay when using a solar simulator. A UVA stimulator is also widely used to evaluate phototoxicity in various test substances. Consequently, we identified the applicability of a UVA simulator to the ROS assay for photoreactivity. In this study, we tested 60 pharmaceutical substances including 50 phototoxins and 10 non-phototoxins to predict their phototoxic potential via the ROS assay with a UVA simulator. Following the ROS protocol, all test substances were dissolved in dimethyl sulfoxide or sodium phosphate buffer. The final concentration of the test solutions in the reaction mixture was 20 to $200{\mu}M$. The exposure was with $2.0{\sim}2.2mW/cm^2$ irradiance and optimization for a relevant dose of UVA was performed. The generation of ROS was compared before and after UVA exposure and was measured by a microplate spectrophotometer. Sensitivity and specificity values were 85.7% and 100.0% respectively, and the accuracy was 88.1%. From this analysis, the ROS assay with a UVA simulator is suitable for testing the photoreactivity and estimating the phototoxic potential of various test pharmaceutical substances.

• Restorative Dentistry and Endodontics
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• v.24 no.1
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• pp.40-48
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• 1999

This study was designed to examine the specificities of the designed primers for F. nucleatum and F. necrophorum and to compare the PCR results using clinical samples with those of the anaerobic culture method. F. nucleatum and F. necrophorum spp. are frequently isolated in infected root canals, and they are related to periapical diseases. F. nucleatum(VPI 10197) and F. necrophorum(ATCC 25286) were used as references for PCR reaction, and thirty five teeth with one canal and periapical lesion were used. The samples were cultured anaerobically and identified using Rapid ID 32A(BioMerieux Vitek, Inc., France) as biochemical battery. In the GenBank database, species-specific PCR primers(nuc1/nuc2 primers for F. nucleatum and nec1/nec2 primers for F. necrophorum) were designed from the 16S ribosomal DNA(rDNA) sequences of F. nucleatum(accession number M58683) and F. necrophorum(accession number AF044948). PCR procedures of F. nucleatum(VPI 10197) and F. necrophorum (ATCC 25286) were simulated on a computer software. Amplify(v.1.2${\beta}$ for Macintosh). 820 bps and 817 bps of nucleotides were expected, respectively. Using extracted DNAs with QiaAmp tissue kit(Qiagen co., Germany), PCR was done. The results were as follows : 1. The nuc1/nuc2 primers produced an amplicon of 820 bps and the nec1/nec2 primers produced an amplicon of 817 bps. 2. The nuc1/nuc2 primers and the nec1/nec2 primers were specific and did not react with species other than the designated ones(i.e. nuc1/nuc2 primers did not produce amplicons for F. necrophorum, and vice versa.). And the PCR products of Porphyromonas endodontalis(ATCC 35406), Porphyromonas gingivalis(ATCC 33277), Prevotella intermedia(ATCC 25611), and Prevotella nigrescens(ATCC 33563), frequently isolated in infected root canals and periapical lesions, were not amplified by the primers specific for Fusobacterium nucleatum and Fusobacterium necrophorum. 3. This method utilizing PCR could detect F. nucleatum and F. necrophorum in clinical samples, while anaerobic culture method could detect neither.

• Korean Journal of Plant Taxonomy
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• v.43 no.2
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• pp.81-89
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• 2013

In general, studies of aerial parasitic plants known collectively as mistletoe have been carried out to investigate their ecological and agricultural characteristics. However, with the recently increased level of interest in medicinal resources, research on different types of Korean mistletoe has also increased. This study was carried out to review the work on the taxonomy and ecology of Korean mistletoe in preparation for the industrial use of these plants in the future. Mistletoe types are flowering plants belonging to Santalales, which exist in the form of parasites on the branches of trees or shrubs. In Korea, five taxa of four genera in two families of mistletoe exist: Viscum coloratum (Komarov) Nakai f. coloratum, Viscum coloratum (Komarov) Nakai f. rubroaurantiacum (Makino) Kitagawa and Korthalsella japonica (Thunb.) Engl. in Santalaceae, along with Loranthus tanakae Franch. et Sav. and Taxillus yadoriki (Sieb. ex Maxim.) Danser in Loranthaceae. As taxonomic studies of these species remain insufficient and given that the distribution ranges of these species are very wide, further observations pertaining to the morphological variations in each species are necessary. The distribution of mistletoes is known to be determined by the host specificity, the interval between the hosts, the environmental condition, the habits of the host plant, the eating characteristics of mediators in the area, and their habitat selection features.

• Journal of Korean Society of Forest Science
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• v.69 no.1
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• pp.42-50
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• 1985

The vegetation of common Cryptomeria (Cryptomeria japonica D. Don) stands planted in Chonnam- province was investigated to obtain the fundamental informations for evaluation of suitable site and the improvement of managing method of the Cryptomeria stands in this region. The results investigated were summarized as follows; 1) The growth condition of common Cryptomeria planted 21-25-year-old stands was similar that of same species growing in Akidaken-district in Japan, while the growth condition of 51-58-year-old stands was not so good as that of Japanese. 2) Total number of plant species was 256. The number of floristic composition varied in the range of 42-99- species, which should be mere than those of Japanese. 3) The investigated sites were located in temperate southern part and in subtropic northern part of Korean peninsular. The types of understory vegetation were classified from I to IV class. 4) From the results of high max. possible diversity (H' max), and dominance (1-J') and from the low simple dominance (${\lambda}$) and evenness (J'), It could be concluded that vegetation was relatively in evenness. 5) From the low percent similarity, the specificity among the stands could be evaluated as considerable. 6) After the index of Morista, the 8th stand in Chang sung showed the generalized vegetation, while the 12th stand in Chang hung showed the specialized vegetation. 7) From the low values of Sneath-Sokal distance, the similarity among the stands investigated appeared very high.

• Biomedical Science Letters
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• v.25 no.4
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• pp.426-430
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• 2019

Currently, molecular diagnostic assays based on nucleic acid amplification tests have been shown to effectively detect mycobacterial infections in various types of specimen, however, variable sensitivity was shown in FFPE samples according to the kind of commercial kit used. The present study therefore used automated PCR-reverse blot hybridization assay (REBA) system, REBA Myco-ID HybREAD 480^®, for the rapid identification of Mycobacterium species in various types of human tissue and compared the conventional one-tube nested-PCR assay for detecting Mycobacterium tuberculosis (MTB). In conventional nested-PCR tests, 25 samples (48%) were MTB positive and 27 samples (52%) were negative. In contrast, when conducted PCR-REBA assay, 11 samples (21%) were MTB positive, 20 samples (39%) were NTM positive, 8 samples (15%) were MTB-NTM double positive, and 13 samples (25%) were negative. To determine the accuracy and reliability of the two molecular diagnostic tests, the one-tube nested-PCR and PCR-REBA assays, were compared with histopathological diagnosis in discordant samples. When conducted nested-PCR assay, 10 samples (59%) were MTB positive and seven samples (41%) were negative. In contrast, when conducted PCR-REBA test, three samples (17%) were MTB positive, 10 samples (59%) were NTM positive and four samples (24%) were negative. In conclusion, the automated PCR-REBA system proved useful to identify Mycobacterium species more rapidly and with higher sensitivity and specificity than the conventional molecular assay, one-tube nested-PCR; it might therefore be the most suitable tool for identifying Mycobacterium species in various types of human tissue for precise and accurate diagnosis of mycobacterial infection.

• Parasites, Hosts and Diseases
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• v.58 no.2
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• pp.153-159
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• 2020

The chigger mite Leptotrombidium sialkotense is one of the 6 main vectors of scrub typhus in China. Before present study, L. sialkotense was found in some parts of Hunan province, China with a narrow geographical distribution. During field investigation 2016-2017, we found L. sialkotense in Jingha, southern Yunnan, China. Of 15 small mammal host species, L. sialkotense were collected from 6 species of the hosts. Rattus brunneusculus was a dominant host of L. sialkotense, from which 98.3% of the mites were collected. The chigger mite showed a relatively high infestation prevalence (P_M =11.7%) and mean abundance (MA=0.5) in comparison with the rest 5 host species. These results reveal a certain host specificity of L. sialkotense to a rat R. brunneusculus. The mite L. sialkotense showed an aggregated distribution on the host (P<0.05). A positive correlation observed between L. sialkotense and the body length of hosts. There was a positive interspecific association between L. sialkotense and 2 other dominant vectors, L. deliense and L. scutellare.