• ALGAE
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• v.24 no.2
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• pp.105-110
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• 2009

The rDNAs of figh-killing dinoflagellates Cochlodinium polykrikoides and Karlodinium veneficum were detected from the East China Sea by species-specific real-time PCR probes. Sequence analysesusing the partial ITS sequences from the real-time PCR products showed identical sequences with C. Polykrikoides and K. veneficum, respectively and low expectation values (E-value) of less than 1e-5 suggesting the presence of these organisms in the East Ching Sea shelf water that flows into the Tsushima Strait and the Yellow Sea.

• Korean Journal of Ecology and Environment
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• v.52 no.1
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• pp.28-39
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• 2019

This study was conducted to elucidate microplastics detection of freshwater ecosystems in Geum river. Samples are collected at 6 points in freshwater, 5 points in fishes. Freshwater was sampled 100 L per each points and fish species were Opsariichthys uncirostris amurensis, Hemibarbus labeo, Pseudogobio esocinus, Zacco platypus, Micropterus salmoides and Cyprinus carpio. FTIR analyis was adopted to identify microplastic types. Extracted microplastics were PES (polyester), PE (polyethylene), PP (polypropylene), PET (polyethylene terephthalate), PVC(Polyvinyl chloride) in freshwater, and PES, PE, PP, PET, PVC in fishes. Our results were expected to be used basic research information for further study in microplastics of freshwater ecosystems.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.132-133
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• 2003

The marine dinoflagellate genus Alexandrium comprise PSP producing A. acatenella, A. angustitabuzatum, A. catenella, A. fundyense, A. minutum, A. ostenfezdii, A. tamiyavanichii and A. tamarense. In monitoring toxic Alexandrium, rapid and exact species identification is one of the significant prerequisite work, however we have suffered confusion of species definition in Alexandrium. To surmount this problem, we chose DNA probing, which has long been used as an alternative for conventional identification methods, primarily relying on morphological approaches using microscope in microbial field. Oligonucleotide DNA probes targeting rRNA or rDNA have been commonly used in diverse studies to detect and enumerate cells concerned as a culture-indetendent powerful tool. Despite of the massive literature on the HAB species containing Alexandrium, application of DNA probing for species identification and detection has been limited to a few documents. DNA probes of toxic A. tamarense, A. catenella and A. tamiyavanichii, and non-toxic A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax were designed from LSU rDNA D1-D2, and applied to whole cell-FISH. Each DNA probes reacted only the targeted Alexandrium cells with very high species-specificity within Alexandrium. The probes could detect each targeted cells obtained from the natural sea water samples without cross-reactivity. Labeling intensity varied in the growth stage, this showed that the contents of probe-targeted cellular rRNA decreased with reduced growth rate. Double probe TAMID2S1 achieved approximately two times higher fluorescent intensity than that with single probe TAMID2. This double probe did not cross-react with any kinds of microorganisms in the natural sea waters. Therefore we can say that in whole-cell FISH procedure this double DNA probe successfully labeled targeted A. tamiyavanichii without cross-reaction with congeners and diverse natural bio-communities.

• Development and Reproduction
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• v.18 no.2
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• pp.99-106
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• 2014

Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival. However, the knowledge of the development of immune system in fish is limited and in demand now. In vertebrates, recombination-activating gene 1 (RAG-1) and immunoglobulin M (IgM) have been considered as very useful markers of the physiological maturity of the immune system. In this study, the expression of the both genes was assessed throughout the early developmental stages of olive flounder larvae (5-55 dph) and used as markers to follow the development of immune system. RAG-1 and IgM mRNA expression was detectable at 5 dph and remained so until 55 dph. These patterns of expression may suggest that the olive flounder start to develop its function around 5 dph. Tissue distribution was found that both genes mRNAs are only expressed in the immune-related organ such as spleen, kidney and gill. The early detection of IgM mRNA led to the investigation of its presence in oocytes. Both RAG-1 and IgM mRNA transcripts were detected in unfertilized oocytes, suggesting that they are maternally transferred. The biological significance of such a phenomenon remains to be investigated.

• Journal of fish pathology
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• v.20 no.3
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• pp.307-313
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• 2007

In this study, a QCM biosensor was made to detect Edwardsiella tarda (E. tarda) using a specific antibody. A 9 MHz AT-cut piezoelectric wafer layered with two gold electrodes of 5mm diameter had a reproducibility of 0.1 Hz in frequency response and was used as the transducer of the QCM biosensor. Self assembled layer (SAM) was conformed on a quartz crystal by treating with 3-mer-captopropionic acid (MPA) and activated with N-ethyl-N'-(3-dimethyl-aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The resulting NHS group was further converted to hydrazide by the reaction with hydrazine. Aldehyde group was introduced into the carbohydrate moiety of anti-E. tarda antibody by the reaction with periodic acid and was used to immobilise the antibody through the reaction with hydrazide group on the electrode surface. A baseline was established in the presence of phosphate-buffered saline (PBS) and a resonant frequency (F1) was measured. Sample was added to the sensor surface and second resonant frequency (F2) was measured after unbound substances were washed out with PBS several times. Finally, the frequency shift (ΔF) representing the mass change was calculated by subtracting F2 from F1. After adding the oxidized anti-E. tarda antibody to the electrode surface containing hydrazide group, frequency shift of 288.811.4 Hz (mean S.E) was observed, thus proving that considerable amount of antibody was immobilized. In the immunoassay test, the frequency shift of 1877.75 Hz, 580.67 Hz, 221.39 Hz, 7.671.83 Hz (mean S.E) were observed at doses of 1000, 500, 100, 50 g of bacterial cells, respectively. It was also demonstrated that the prepared sensor chip was stable enough to withstand repeated surface regeneration with 0.2 M Tris-glycine and 1 % DMSO, pH 2.3 more than ten times.

• Parasites, Hosts and Diseases
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• v.21 no.2
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• pp.187-192
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• 1983

In order to clarify infestation Pattern for the encysted larvae of digenetic trematodes from fresh-water fishes, this survey was carried out from March to September, 1983. A total of 380 fishes of 32 species wore collected with netting at the three reaches, upper, middle and lower in Mangyeong riverside area. After the fishes were dissected into small scraps, they were pressed under cover glass and examined-for the presence of those of digenetic trematodes with a microscope. The resillts obtailled were as follows; Out of a total of 380 knishes inspected, 320 fishes(84%) from 31 species were found positive with, digenetic trematode metacercariae; more than 10 species of the metacercariae Ivere detected in Pseuderasbora parva; .Gnnthepegen mtajimae, Microphysogokio yaluensis, Cultriculus eigenmanni and Gnnthopogon coreanus (more than 8 species) ; Aphyocypris chinensis (8 species) and etc. respectively. Clonorchis sinensis metacercariae were found positive from 93 fishes (25%) from 12 species and detection rates in other species of digenetic trematode metacercariae from various fishes were; Exorchis oviformis, 261 fisles (57%) from 28 species; Cyathocotyle criensalis, 47 fishcs (12%) from 12 species; Metorchis orientalis, 21 fishes (6%) from 12 species; Metagonimus yokogawai, 164 fishes (43%) from 26 species; Pseudesorchis major, 71 fishes (19%) from 18 species; Metacercaria haiegawai, 77 fishes (20%) from 25 species; Centrocestus armatus, 24 fishes (6%) from 7 species; Echinochasmus japonicus, 2 fishes (0.5%) from 2 species, and unidentified species, 34 fishes (9%) from 15 species respectively. The sums of average number of the encysted larvae of all species found in fish body/gram showed 83 in P.parva, Cobitis taenia (74.2), A. chinensis (28.5), Pseudoperilampus uyekii (26.6), G. majimae (19.6) and etc. respectively and the average peak number of each metacercaria in fish body/gram showed 21.7 C. sinensis, 245. ovifcrmis, 15, 3 M, crientalis and 6.1 5. japonicus in P. parva. 42.7 C. orientalis and 25.1 M. yohogawai in C. taenia; 8.3 C. armatus and 8.3 M. hasegawai in P. uyekii. 6.3 P. major in Carassius carassius, and 2.9 unidentified species in G, niajimae respectively.

• Environmental Analysis Health and Toxicology
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• v.22 no.1 s.56
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• pp.19-26
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• 2007

Accumulation of tributyltin (TBT) in serum, liver and muscle of olive flounder (Paralichtys olivaceus) was investigated in a 60-d static renewal exposure at $0.1{\mu}g$ TBT/L. Tributyltin accumulated rapidly from 83 ng Sn/g to 2,227 ng Sn/g on a wet weight basis in the serum of the olive flounder and to a greater extent than in the other tissues. The accumulated TBT concentrations in tissues were in the order of serum>liver>muscle on wet or dry-weight basis. High concentrations of dibutyltin (DBT: 990 ng Sn/g dry wt) and monobutyltin (MBT: 141 ng Sn/g dry wt), degradation products of TBT were found in liver of olive flounder at the end of exposure. On the other hand, DBT and MBT was below the detection limits in muscle during the exposure, and only low concentration of DBT (56 ng Sn/g) were detectefd in serum. Butyltin compounds were also quantitatively determined in feral fine-spotted flounder (Pleuronichthys cornutus) collected from Gwangyang Bay as one of polluted area and Sori Island as a reference site. All three butyltin compounds were detected from the fine-spotted flounder from Gwangyang Bay up to 3,107 ng Sn/g of total butyltins in liver, while 120 ng Sn/g of total butyltin concentration was found in the liver of fish from Sori Island.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1672-1678
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• 2009

We studied the effects of aged total petroleum hydrocarbons (aged TPH) on the survival of allochthonous diesel-degrading Rhodococcus sp. strain YS-7 in both laboratory and field investigations. The aged TPH extracted from a crude-oil-contaminated site were fractionized by thin-layer chromatography/flame ionization detection (TLC/FID). The three fractions identified were saturated aliphatic (SA), aromatic hydrocarbon (AH), and asphaltene-resin (AR). The ratio and composition of the separated fractions in the aged TPH were quite different from the crude-oil fractions. In the aged TPH, the SA and AH fractions were reduced and the AR fraction was dramatically increased compared with crude oil. The SA and AH fractions (2 mg/l each) of the aged TPH inhibited the growth of strain YS-7. Unexpectedly, the AR fraction had no effect on the survival of strain YS-7. However, crude oil (1,000 mg/l) did not inhibit the growth of strain YS-7. When strain YS-7 was inoculated into an aged crude-oil-contaminated field and its presence was monitored by fluorescent in situ hybridization (FISH), we discovered that it had disappeared on 36 days after the inoculation. For the first time, this study has demonstrated that the SA and AH fractions in aged TPH are more toxic to an allochthonous diesel-degrading strain than the AR fraction.

• Journal of Microbiology and Biotechnology
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• v.28 no.11
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• pp.1823-1833
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• 2018

Fluorescence in-situ hybridization (FISH) is a common and popular method used to investigate microbial communities in natural and engineered environments. In this study, two specific 16S rRNA-targeted oligonucleotide probes, CLZ and KCLZ, were designed and verified to quantify the genus Clostridium and the species Clostridium kluyveri. The optimal concentration of hybridization buffer solution for both probes was 30% (w/v). The specificity of the designed probes was high due to the use of pellets from pure reference strains. Feasibility was tested using samples of Chinese liquor from the famed Luzhou manufacturing cellar. The effectiveness of detecting target cells appears to vary widely in different environments. In pit mud, the detection effectiveness of the target cell by probes CLZ and KCLZ was 49.11% and 32.14%, respectively. Quantitative analysis by FISH technique of microbes in pit mud and fermented grains showed consistency with the results detected by qPCR and PCR-DGGE techniques, which showed that the probes CLZ and KCLZ were suitable to analyze the biomass of Clostridium spp. and C. kluyveri during liquor fermentation. Therefore, this study provides a method for quantitative analysis of Clostridium spp. and C. kluyveri and monitoring their community dynamics in microecosystems.

• Journal of fish pathology
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• v.9 no.1
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• pp.33-40
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• 1996

A filamentous form of mollicute-like bacterium was detected during a routine health survey of wild adult kuruma shrimp Penaeus japonicus. The kuruma shrimp were native to Japan and were imported to Korea. The histology showed no pathological changes. Transmission electron microscopy revealed extensive infections of hepatopancreatic epithelial cell by a pleomorphic, filamentous intracellular bacterium. The filamentous bacterium was of about 60 nm in diameter and 300 nm to more than $1{\mu}m$ in length. Teh morphology of bacteria were filamentous and branched with terminal blebs, or knobs, on the branches. They lacked the cell wall, and were bounded by the plasma membrane. They contained typical prokaryotic ribosomes and fibrillar DNA-like strands. No additional internal structure has been observed. They are considered to be mollicutes, based upon the morphological appearance and upon the cellular location.