• Journal of Korean Society of Environmental Engineers
• /
• v.29 no.8
• /
• pp.904-908
• /
• 2007

The protozoan pathogen Giardia lamblia has been major cause of waterborne enteric disease. In this study, we tried to identify G. lamlbia of human infectious species and to detect viable C. lamblia in river water samples including three sites of Han River mainstream and an its creek using PCR and RT-PCR technique. The PCR/RT-PCR methods were performed by using giardin primer based on the giardin gene targeting ventral disk of Giardia. Sensitivity testing in the DNA/RNA extraction and PCR/RT-PCR amplification steps showed that it was possible to detect a single cyst of G. lamblia and viable G. lamblia. The PCR/RT-PCR methods were compared with immunofluorescence(IF) assay by analyzing 48 samples collected from the mainstream water and the creek water. The mean concentration of the total cysts were 6.3 cysts/10 L(arithmetic mean, n = 48) and the positive detection rate were 62.5%(30/48). And the mean concentration of the cysts excluding empty cysts were 4.5 cysts/10 L and the positive detection rate were 52.1%(25/48). It resulted that 24 of 48 samples included Giardia lamblia by PCR assay and 10 of 48 samples included viable G. lamblia by RT-PCR assay. It resulted that the PCR/RT-PCR technique would be available to river water samples with low concentration of Giardia cysts. And it could support the Korean protozoan standard method, which provides useful information for species and viability.

• Biomedical Science Letters
• /
• v.27 no.4
• /
• pp.255-263
• /
• 2021

Human Astrovirus (HuAstV) is an important gastrointestinal pathogen that is frequently reported worldwide. Monitoring of contaminated groundwater has been suggested since HuAstV is transmitted through the fecal-oral route. This study developed a test method based on conventional reverse transcription (RT)-nested polymerase chain reaction (PCR) that involves SL^® non-specific reaction inhibitor for unknown non-specific amplification taking place in the groundwater environment. An optimal method for detecting HuAstV in groundwater sample through analysis and comparison against conventionally reported method was also suggested. The developed method enabled the production of nested PCR amplicon of 630 nt, which is a sufficient length for similarity analysis based on sequencing and genotyping. Amplicons suspected to be HuAstV were amplified in two out of the twenty groundwater samples collected in Korea, presenting 99.77% and 99.73% similarity against HuAstV 1 strain lhar/2011/kor (JN887820.1) in sequencing, respectively. These amplicons were identified as HuAstV 1.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.32 no.1
• /
• pp.35-41
• /
• 2003

The study was conducted to develope a rapid method for the detection of Yersinia enterocolitica in spring water and vegetables via multiplex polymerase chain reaction (PCR) technique using ail, yst, uirF and subgenus-specific Y16S primers. Specificity and sensitivity of multiplex PCR and application of best primers for the detection of Y. enterocolitica from spring water and vegetables were investigeted. Y. enterocolitica ATCC 27729 strains gave 356 bP and 200 bp (Y16S) and 134 bp (yst) bands. but Y. enterocolitica ATCC 9610 and ATCC 23715 strains gave 200 bp and 134 bp bands.In the meanwhile, non-pathogenic Yersinia species, such as Y. frederikseni, Y. inter-media, Y. kristenseni and Y. pseudotuberculosis gave only single 200 bp band, and other bacteria including Escherichia coli O157:H7 ATCC 25392, Shigella dysenteri. Staphylococcu aureus ATCC 25923 and Listeria mo-nocytogenes ATCC 19111 did not show any bands. Among primers, yst and Y16S primer showed the best sensitivity. Seven CFU/mL Y. enterocolitica cells could be detected with yst and Y16S primers and the sensitivity was significantly improved by the further 2nd PCR after 38 cycles of first PCR amplication. Spring water, cabbage and mushroom were inoculated with Y. enterocolitica to determine the sensitivity of multiplex-PCR for the rapid detection of Y. enterocolitica. Multiplex-PCR assay could detect 7 or 70 cells in spring water and vegetables using whole cell lysate with repeating PCR amplication.

• Journal of Aquaculture
• /
• v.13 no.2
• /
• pp.129-135
• /
• 2000

Genetic variabilities of local populations of Anthocidaris crassispina and Hemicentrotus pulcherrimus were analysed by random amplified polymorphic DNA (RAPD) technique with phenotypic variabilities in timing of gonadal maturation, reproductive output and size of individuals in Cheju. H, pulcherrimus, collected from 4 locations during March 1997, indicated that Daepo individuals were significantly smaller than that at Hamdok, Wimi site A and Wimi site B (ANOVA, p>0.001). Gonodal-somatic index (GSI) of the Wimi site B population was significantly higher than that of three other locations (ANOVA, P>0.0001). An ANOVA test conducted on test size of A. crassispina, harvested from six different locations of Cheju during June 1997, indicated that the size of individuals from Pophwan was significantly smaller (P>0.0001) than that from five other locations. GSIs of urchin m Wimi and Hanlim were significantly higher than that from Pohwan and Oedo (ANOVA, p>0.0001). Genetic similarity, calculated from k13 primer analysis of total DNA, among the six different populations of A. crassispina varied from 0.67 to 0.92, and the values for H. pulcherrimus from 0.60 to 0.73; thus there was no genetic variation among different populations of the same species. Therefore, the populations are genetically homologous and the observed phenotypic variabilities were possibly associated with water temperature and food.

• The Journal of Korean Academy of Prosthodontics
• /
• v.61 no.3
• /
• pp.179-188
• /
• 2023

Purpose. The purpose of this study was to evaluate the effect of surface treatments on the shear bond strength of two types of zirconia (3-TZP and 5Y-PSZ) with resin cement. Materials and methods. Two different types of zirconia specimens with a fully sintered size of 14.0×14.0×2.0 mm³ were prepared, polished with 400, 600, and 800 grit silicon carbide paper, and buried in epoxy resin. They were classified into four groups each control, sandblasting, primer, and sandblasting & primer. Cylindrical resin adhered to the surface-treated zirconia with resin cement. It was stored in distilled water (37℃) for 24 hours, and a shear bond strength test was performed. The normality of the experimental group was confirmed with the Kolmogorov-Smirnov & Shapiro-Wilk test. The interaction and statistical difference were analyzed using a two-way ANOVA. A post-hoc analysis was performed using Dunnett T3. Results. As a result of two-way ANOVA, there was no significant difference in shear bonding strength between zirconia types (P > .05), but there was a significant correlation in the sandblasting, primer, and alumina sandblasting & primer group (P < .05). Dunnett T3 post-test showed that, regardless of the type of zirconia, shear bonding strength was sandblasting & primer > Primer > sandblasting > control group (P < .05). Conclusion. There was no difference in shear bond strength between the types of zirconia. The highest shear bond strength was shown when the mechanical and chemical treatments of the zirconia surface was performed simultaneously.

• Journal of environmental and Sanitary engineering
• /
• v.15 no.4
• /
• pp.105-113
• /
• 2000

We have isolated 65 strains(2.0%) of Y. enterocolitica among 3,219 water samples from 380 spring water sites in Seoul from 1994 to 1999. The biochemical characteristics of isolated strains revealed that TSI was A/A, urea, M.R.($37^{\circ}C$), nitrate, motility($37^{\circ}C$), sorbitol, maltose, manitol, arabinose, mannose, trehalose, xylose were positive(100%) and H$_2$S, arginine, lysine, oxidase, citrate, V.P.($37^{\circ}C$), DNase, motility($37^{\circ}C$), dulcitol, adonitol, lactose and raffinose were negative(100%). In in vitro virulence test, positive rate of AAG and CRMOX were 9.2% and 4.6%, respectively. However in the virulence gene detectable gene detectable test by multiplex-PCR using ail, yst, virF genes, 65 strains were all negative, meaning that Y.enterocolitica strains from domestic spring water were not detected for the virulence. Otherwise, mutiplex-PCR which using ail, yst and subgenus-specific primer pair was the best for identifying the virulence of Y. enterocolitica.

• Journal of Dental Rehabilitation and Applied Science
• /
• v.22 no.2
• /
• pp.193-202
• /
• 2006

Objective To evaluate the influence of water storage on the mechanical properties of dental adhesives over 1 and 3 months. Materials and Methods Adhesive resin sheets were prepared by pouring either All-bond 2(AB), Clearfil SE Bond(SE) into a mold measuring $15{\times}15{\times}0.9mm$. After solvent in primer evaporation, the adhesives were light-cured and removed from the mold and divided in two pieces, trimmed to hourglass shape that were used to determine the micro-tensile strength(MTS). Another hourglass shaped metal mold measuring $2.0{\times}1.5mm$ in cross-section area was made to determine the Young's modulus(E). Adhesive specimens for Young's modulus(E) were prepared in the same method. Specimens were stored at $37^{\circ}C$ in distilled water and tested after 1 and 3 months. The data were analyzed by one-way ANOVA and Tukey's test. Results Water storage significantly decreased the micro-tensile strength(MTS) of AB and SE specimens after 1 and 3 months(P<0.05). The Young's modulus(E) were also decreased after water storage for 1 and 3 months, but statistically not significant in each group of AB and SE group respectively. Conclusions Long-term exposure of adhesive resin to water can cause reduction of mechanical properties. It may compromise resin/dentin bonds and affect longevity of restorations.

• Journal of fish pathology
• /
• v.31 no.2
• /
• pp.93-99
• /
• 2018

Edwardsiella tarda predominantly causes edwardsiellosis in fish at high temperature, but is rarely isolated from water when water temperature is low. However, E. tarda is viable but nonculturable (VBNC) in low water temperature, but it can be revived when water temperature rises and cause disease to fish. Therefore, in order to prevent disease, it is very important to identify pathogens that are in the VBNC state in environmental water. In this study, E. tarda cells in the VBNC state were detected by the ethidium monoazide (EMA)-PCR method using the low-temperature oligotrophic sea water microcosm obtained by inoculation of E. tarda at a concentration of $10^8CFU/ml$. In order to distinguish between live and dead bacteria in E. tarda, each sample was treated with EMA at different concentrations, photoactivated with a 500 W halogen lamp, and PCR was performed with E. tarda specific primer. At the concentration of $10^7CFU/ml$ bacterium, DNA amplification was observed only in the live cells when treated with $60{\mu}g/ml$ of EMA, and smaller amounts of live cells could be distinguished from dead cells by adjusting the EMA concentration. In addition, the VBNC cells of E. tarda in the oligotrophic low temperature seawater microcosm were estimated to be in the range of $10^4{\sim}10^5CFU/ml$ by EMA-PCR. Therefore, it is possible to detect VBNC cells that will act as potential pathogens in environmental water using EMA-PCR method, and quantitative confirmation using concentration change is also possible.

• Biomedical Science Letters
• /
• v.27 no.1
• /
• pp.35-43
• /
• 2021

Human Sapovirus (HuSaV) is one of the major causes of acute gastroenteritis in humans, and it is used as a molecular diagnostic technique based on polymerase chain reaction (PCR) from humans, food, shellfish, and aquatic environments. In this study, the HuSaV diagnosis technique was used in an aquatic environment where a number of PCR inhibitors are included and pathogens, such as viruses, are estimated to exist at low concentration levels. HuSaV-specific primers are improved to detect 38 strains registered in the National Center for Biotechnology Information (NCBI). The established optimal condition and the composition, including the RT-nested PCR primers and SL® Non-specific reaction inhibitor, were found to have 100 times higher sensitivity based on HuSaV plasmid than the previously reported methods (100 ag based on HuSaV plasmid 1 ng/μL). Through an artificial infection test, the developed method was able to detect at least 1 fg/μL of HuSaV plasmid contaminated with total nucleic acid extracted from groundwater. In addition, RT-nested PCR primer sets for HuSaV detection can react, and a positive control is developed to verify false positives. This study is expected to be used as a HuSaV monitoring method in the future and applied to the safety response to HuSaV from water environments.

• Advances in environmental research
• /
• v.8 no.1
• /
• pp.71-84
• /
• 2019

Effective waterproofing of structures was a compulsory constraint to avoid leaks and dampness or humidity in walls, ceilings, roofs underground tank and underground room. Traditionally used methods of roof waterproofing were bitumen with tinny seared clay tiles are very troublesome, overwhelming time and involving high labor cost. These waterproofing methods are not allocation the purpose due to their intrinsic disadvantages. Prepackaged polymer modified slurries (PPPMS) are now attainment the vogue and easy to use, easily available in the market, cheaper in cost and more workable than the traditional methods of waterproofing. An experimental study has shown that prepackaged polymer modified slurries (PPPMS) are superior in cost and performance to as a roof water proof coatings. Bituminous coatings were mixed with water and different combination of prepackaged polymer modified slurries and primer respectively, to find optimum coverage underneath worst atmospheric conditions. Every specimen of different proportioned was applied on plane roofs and through the passage of time, their performance was checked, assessed and associated with each other. The roof of approximately 40000 ft² area of prepackaged polymer modified slurries was used will give us hundred percent result (no water seepage or no water absorption) therefore no complaints as compare to roofs area of approximately 24000 ft² bituminous coating was used for waterproofing they have shown the result of 30 to 40 percent water seepage. This result shows that prepackaged polymer modified slurries were two times cheaper than bituminous coating. Comparing an equal number of surfaces coated with a polymer modified prepackaged mortar and bitumen the prepackaged polymer modified slurries (PPPMS) showed excellent performance, ease of application and low bitumen coating cost.