• Journal of Korean Society on Water Environment
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• v.34 no.4
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• pp.375-381
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• 2018

The monitoring of phytoplankton biomass and community structure is essential as a first step to control the harmful cyanobacterial blooms in freshwater systems, such as seen in rivers and lakes, due to the process of eutrophication and climate change. In order to quantify the biomass of phytoplankton with a wide range in size and shape, the measurement of cell biovolume along with cell density is required for a comprehensive review on this issue. However, most routine monitoring programs preserve the gathered phytoplankton samples before analysis using chemical additives, because of the constraint of time and the number of samples. The purpose of this study was to investigate the cell biovolume change characteristics of six cyanobacterial species, which are common bloom-causing cyanobacteria in the Nakdong River, after the preservation with Lugol's iodine solution. All species showed a statistically significant difference after the addition of Lugol's iodine solution compared to the live cell biovolume, and the cell biovolume decreased to the level of 34.0 ~ 56.3 % at maximum in each species after the preservation. The nonlinear regression models for determining the shrinkage ratio by a preservation period were derived by using the cell biovolume measured until 180 days preservation of each target species, and the equation to convert the cell biovolume measured after preservation for a certain period to the cell biovolume of viable cell was derived using that formula. The conversion equation derived from this study can be used to estimate the actual cell biovolume in the natural environment at the time of sampling, by using the measured biovolume after the preservation in the phytoplankton monitoring. Moreover this is expected to contribute to the final interpretation of the water quality and aquatic ecosystem impacts due to the cyanobacterial blooms.

• Journal of Microbiology and Biotechnology
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• v.23 no.3
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• pp.289-296
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• 2013

Cryptomonads are unicellular, biflagellate algae. Generally, cryptomonad cells cannot be preserved well because of their fragile nature, and an improved methodology should be developed to identify cryptomonads from natural habitats. In this study, we tried using several cytological fixatives, including glutaraldehyde, formaldehyde, and their combinations to preserve field samples collected from various waters, and the currently used fixative, Lugol's solution was tested for comparison. Results showed that among the fixatives tested, glutaraldehyde preserved the samples best, and the optimal concentration of glutaraldehyde was 2%. The cell morphology was well preserved by glutaraldehyde. Cells kept their original color, volume, and shape, and important taxonomic features such as furrow/gullet complex, ejectosomes, as well as flagella could be observed clearly, whereas these organelles frequently disappeared in Lugol's solution preserved samples. The osmotic adjustments and buffers tested could not preserve cell density significantly higher. Statistical calculation showed the cell density in the samples preserved by 2% glutaraldehyde remained stable after 43 days of the fixation procedure. In addition, DNA was extracted from glutaraldehyde preserved samples by grinding with liquid nitrogen and the 18S rDNA sequence was amplified by PCR. The sequence was virtually identical to the reference sequence, and phylogenetic analyses showed very close relationship between it and sequences from the same organism. To sum up, the present study demonstrated that 2% unbuffered glutaraldehyde, without osmotic adjustments, can preserve cryptomonads cells for identification, in terms of both light microscopy and phylogenetic analyses based on DNA sequences.

• Journal of fish pathology
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• v.19 no.1
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• pp.7-15
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• 2006

Amyloodinium sp. was found on the gills of mullet (Mugil cephalus) cultured on land. No external symptoms in the diseased fish were found except decoloration of the gills. In fresh preparations of the gills the parasites were opaque round or oval shape with a bright nucleus and 43.5 ㎛ (18.2～72.7, n=20) in size. In preparations added a drop of Lugol solution, they were black with the same shapes in fresh preparations and 43.5 ㎛ (n=20) in size. The parasites were stained black and blue in a droplet of Lugol solution and Diff-Quick III solution, respectively and their sizes were a little larger than in wet preparations. After stained with May-Grunwald Giemsa, the parasites appeared granular eosinophlic in the peripheral cytoplasm and granular strong basophilic in the center. In silver impregnated specimens the peripheral granules were negative and the central ones positive. The granules appeared brown in purplish cytoplasm after staining with Lugol solution. The parasites developed by binary division when they were cultivated in filtered seawater at 20℃. Histopathologically severe epithelial hyperplasia and fusion in the gill filaments resulted in clubbing, especially the proximal region of the filament. Epithelial hyperplasia was also found in the basal regions of the gill filaments and some epithelial cells were occasionally detached from the filaments. Some pear-shaped trophonts of the parasites with rhizoid attached on the gill filaments showing hyperplasia of the epithelial cells and mucous cells.

• Korean Chemical Engineering Research
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• v.54 no.1
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• pp.140-144
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• 2016

In this study, the physical factors for amylase production by Arthrobacter sp. were optimized using response surface methodology(RSM). Antarctic microorganism Arthrobacter sp. PAMC 27388 was obtained from the Polar and Alpine Microbial Collection(PAMC) at the Korea Polar Research Institute. This microorganism was confirmed for the excretion of amylase with Lugol's solution. The amylase activity was after flask culture was as low as 1.66 mU/L before optimization. The physical factors including the inoculum volume, the initial culture pH, and the medium volume were chosen to be optimized for the enhanced amylase production. The calculated results using RSM indicate that the optimal physical factors were 2.49 mL inoculum volume, 6.85 pH and 42.87 mL medium volume with a predicted amylase production of 2.84 mU/L. The experimentally obtained amylase activity was 2.50 mU/L, which was a 150% increase compared to the level before optimization.

• Journal of Korean Society on Water Environment
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• v.34 no.2
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• pp.210-215
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• 2018

It is difficult to count colonial cyanobacteria Microcystis cells since the thickness of colonies is constrained by amorphous mucilage, making it impossible to estimate the number of cells. Disaggregation of Microcystis colonies into single cell is needed to improve the accuracy and precision of cell density estimation of naturally collected samples. Uultrasonic treatment method is commonly used owing to the simplicity and immediacy of the procedure. However, amplitude, frequency, and duration of ultrasonic treatment also cause cell loss during the experiment. Optimal ultrasonic treatment has not been standardized yet. Therefore, the objective of this study was to investigate optimal ultrasonic treatment by analyzing cell density and colony numbers. We collected colonial Microcystis from Changnyeong-Haman weir area in Nakdong River during harmful algal boom period from September to October in 2017. Ultrasonic treatment method was applied to disrupt colonies into single cells to enumerate cell density. Among treatment conditions, results from continuously treated for 100 seconds were found to be the optimum to reduce colonies to a suspension of single cell without cell losses under high and low density of Microcystis cells. Lugol iodine fixed cells followed by sonication showed less negative impact of cell damage within the optimal treatment time (100 seconds). Furthermore, disaggregated cells treated by sonication enables microscopic observation more easily since gas vacuoles were collapsed to facilitate sedimentation of cells under the counting chamber for quantitative enumeration of buoyant Microcystis cells.

• Journal of Chest Surgery
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• v.23 no.3
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• pp.537-541
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• 1990

Early esophageal carcinoma is defined as a lesion wherein invasion is confined to the mucosa and submucosa without metastasis to lymph node or other organs. Postoperative 5-year survival rate for early esophageal carcinoma is much superior than advanced carcinoma. Unfortunately, because of the anatomic characteristic of esophagus and absence of specific early symptoms, detection is frequently belated, and advanced disease is present at the time of the initial diagnosis. We experienced 2 cases of early esophageal carcinoma. They complained no specific symptoms. The diagnosis was made by barium esophagogram, esophagofiberscopy with dye staining and endoscopic biopsy. We performed esophagectomy with esophagogastrostomy. All had good postoperative course without any complication. We concluded that the combined use of double contrast radiography, esopagofiberscopy aided by intraluminal staining with Toluidine blue or Lugol`s solution, and endoscopic biopsy is very important in the diagnosis of early esophageal carcinoma in high risk patient group.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.4 no.4
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• pp.363-370
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• 1999

When the first red tide alert by Cochlodinium polykrikoides was alarmed around the Oenarodo coast on Aug. 27, 1997, there co-occurred two chain-forming naked dinoflagellates which were different sized but looked fairly similar. The analyses of 24S rRNA sequences of these species showed that their gene sequences had only 74.9% identity. This low value implies that they are quite different species. After isolation and cultivation of each species, the morphological characteristics were observed. This revealed that the larger species ranging from 20 to 35 ${\mu}m$ was the well known, Cochlodinium polykrikoides and the smaller one ranging from 12 to 25 ${\mu}m$ was Gyrodinium impudicum which had not been reported in Korea. As their 24S rRNA sequences had not been analysed yet, we deposited the sequences in Genbank. At that time of the investigation. the red tide was caused by G. impudicum of which maximum cell counts reached up to 30,000 cells $ml^{-1}$. In this study we describe the morphological characteristics and the behavioral patterns of each species which can be easily observed with light microscope or stereomicroscope. In addition, their morphology transformed by the fixation with Lugol's solution are also characterized. which can help to discriminate each one in the fixed sample.

• Fisheries and Aquatic Sciences
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• v.2 no.2
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• pp.172-175
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• 1999

Relative DNA contents in some harmful algae were measured using DAPI staining and image analysis system. This method was useful to identify some morphologically similar species and isolates from harmful algal blooms (HABs). In exponential phase, Prorocentrum micans had higher relative DNA content (RD) of $1.83\pm0.52$ than any other isolates, followed by Cochlodinium polykrikoides $(1.10\pm0.46)$ Alexandrium tamarense $(0.93\pm0.32)$ Gyrodinium impudicum $(0.56\pm0.17)$, Scrippsiella trochoidea $(0.41\pm0.26)$ and P. minimum$(0.05\pm0.01)$. When they were fixed with Lugol's solution, it was difficult to d,iscern C. polykrikoides from G. impudicum under the light microscope, but the DNA contents were quite different in two species. C. polykrikoides contained about twice as much RD as G. impudicum under the same culture conditions and exponential phase. DAPI­stained DNA feature in C. polykrikodes showed concentrated in the peripheral part of the cell, but in G. impudicum showed a compact structure in the central part. Although A. tamarense and S. trochoidea were morphologically similar under the light microscope, nuclear DNA content of A. tamarense was twice as much as that of S. trochoidea.

• Journal of Aquaculture
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• v.10 no.3
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• pp.227-237
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• 1997

Five species of intertidal clams including Ruditapes philippinarum, Tegillarca granosa, Solen strictus, Heteromacoma irus, and Coecella chinensis were tested for the presence of the protozoan parasite, Perkinsus sp. using fluid thioglycollate medium (FTM) fortified with antibiotics and histological techniques. Each individual clam was placed in a test tube filled with 10ml FTM, placed in totally dark place, and incubated over a week. After incubation the clam tissues were stained with Lugol's iodine solution and examined under a light microscope to find out any hypnospores of Perkensus sp. in the tissues. Cross-sections of the clams were also embedded in paraffin, sliced to 3um, and stained with Harry's hematoxylene and Picro eosine to observe the presence of tomont or trophozoites. Perkinsus sp. were found in the presence of tomont or trophozoites. Perkinsus sp. were found in the tissues of R. philippinarum collected from Kangjin and Wando, along the south coast of Korea. However, Perkinsus sp. was not found in four other species of clams nor R. philippinaurm collected from Kimnyong and Waido in Cheju. A size-dependent Perkinsus sp. infection was found in R. philippinarum collected rom Kangjin and Wando the clams smaller than 15mm in shell width do not exhibit and Perkinsus sp. while other clams greater than 20mm in shell width exhibit almost 100% infection. To determine the number of Perkinsus sp. in the clams, FTM cultured clam tissues were digested with 2M NaOH solution and the number of hypnospores in the tube were counted. The number of hypnospores counted from the tissues indicated that each Manila clam contains 100,000 to 3,500,000 Perkinsus cells or 20,000 to 1,000,000 cells per gram tissue wet weight. The results of cell counts also suggests that such a high occurrence of Perkinsus sp. in the clam may cause mortality, as already reported from other studies of Perkinsus spp.

• Journal of agricultural medicine and community health
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• v.15 no.2
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• pp.107-111
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• 1990

A survey was undertaken to evaluate the present status of intestinal infection in army soldiers. Stool specimen of 417 soldiers fixed by SAF solution were collected from a camp located in area of Nakdong river during the period from August to October of 1990. And these stool samples were examined by formalin-ether sedimentation technique once for helminths and protozoan cysts stained with Lugol's iodine solution. The results obtained in this survey were summarized as follows : l) The overall positive rate of intestinal parasite was 18.0%. 2) The egg positive rate of intestinal helminth was 15.1%, : and 11.5% for Clonorchis sinensis. 5.0% for Metagonimus vokogawai, 1.2% for Ascaris lumbricoides 1.7% for Trichiuris trichiura. 0.2% for Taenia sp. 3) The cyst positive rate of intestinal protozoa was 4.1% ; and 1.4% for Entamoeba coli, 1.9% for Giardia lamblia 0.7% for Entamoeba histolytica, 0.5% for Endolimax nana. 4) Most of samples were positive(85.3%) by single species. 10.7% by two species, 2.7% by three species and 1.3% by four species. 5) Infection rate of intestinal parasites among army soldiers decreased distinctly compared with previous data but it is revealed that the infection rate of Clonorchis sinensis among army soldiers in area of Nakdong river is still high in comparison with ever-reported data. 6) SAF fixatives used in this field survey during summertime was useful to conserve protozoan cyst and helminths ova. Also we could examine stool samples directly by formalin-ether sedimentation technique.