• Korean Journal of Veterinary Service
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• v.38 no.1
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• pp.37-42
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• 2015

The aim of the present study was to compare the non-invasive methods for the diagnosis of H. felis with HpSA kit-based detection method and H. felis-specific PCR assay with dog's stool samples without sacrifice. Male Beagle dogs (n=6) were infected with H. felis ATCC 49179 ($1.0{\times}10^9CFU/dog$) by intra-gastric inoculation two times at 3-day intervals, and the stool specimens of dogs were collected 1, 3, 5, 7, 14, 21 days after infection to submit to HpSA test and H. felis-specific PCR. As the results, the sensitivity of the HpSA and the PCR analysis was 50.0%, 83.3% respectively. Although HpSA test is less sensitive, it could be used for rapid, cheap and easy screening assay for H. felis infection in dog and cats. We suggest that the H. pylori stool antigen kit, HpSA, is useful and effective for monitoring H. felis infection. If HpSA test would be made with H. felis antibodies in the future, its sensitivity could be increased. Also, PCR assay could be successfully used to detect the H. felis in stools. Applying the H. pylori stool antigen kit and PCR assay may be the recommended non-invasive strategy to identify H. felis in dog and cats.

• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1616-1621
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• 2007

We have established a SYBR Green-based realtime PCR method using AnyDirect solution, which enhances PCR from whole blood, for direct amplification of the virA gene of Shigella flexneri and the invA gene of Salmonella typhimurium from human feces without prior DNA purification. When we compared the efficiency of conventional or realtime PCR amplification of the virA and invA genes from the supernatant of boiled feces supplemented with S. flexneri and S. typhimurium in the presence or absence of AnyDirect solution, amplification products were detected only in reactions to which AnyDirect solution had been added. The detection limit of real-time PCR was $1{\times}10^4\;CFU/g$ feces for S. flexneri and $2{\times}10^4\;CFU/g$ feces for S. typhimurium; this sensitivity level was comparable to other studies. Our real-time PCR assay with AnyDirect solution is simple, rapid, sensitive, and specific, and allows simultaneous detection of S. flexneri and S. typhimurium directly from fecal samples without prior DNA purification.

• The Plant Pathology Journal
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• v.28 no.1
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• pp.1-9
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• 2012

In this study, 200 isolates of fluorescent pseudomonads were isolated from different fields of East and West Azarbaijan and Ardebil provinces of Iran. These bacterial isolates were screened on the basis of a dual culture assay, the presence of known antibiotic genes, and their ability to successfully colonize roots and to promote plant growth. Twelve isolates exhibited 30% or more inhibition of mycelia growth of $P.$ $drechsleri$. Genes encoding production of the antibiotics 2,4-diacetylphloroglucinol, phenazine-1-carboxylic acid, and pyoluteorin were detected in some strains but none of the strains possessed the coding gene for production of antibiotic pyrrolnitrin. In an $in$ $vitro$ test for root colonization, the population density on roots of plants treated with most of the above strains was more than 6 $\log_{10}$ CFU $g^{-1}$ roots, with a maximum of 7.99 $\log_{10}$ CFU $g^{-1}$ roots for strain 58A. Most of the strains promoted significant plant growth in comparison to non-treated controls. In green house studies, the percentage of healthy plants in pots treated with strains 58A and 8B was 90.8% and 88.7%, respectively. The difference between these treatments and treatment with the fungicide metalaxyl was not significant.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.310-320
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• 2005

A membrane-based oligonucleotide array was used to detect predominant bacterial species in human fecal samples. Digoxygenin-labeled 16S rDNA probes were generated by PCR from DNA that had been extracted from fecal samples or slurries. These probes were hybridized to an array of 120 oligonucleotides with sequences specific for 40 different bacterial species commonly found in human feces, followed by color development using an alkaline phosphatase-conjugated antibody and NBT /BCIP. Twenty of the species were detected by this method, but E. coli, which was present at $\~$1 $\times 10$^5$ CFU per gram feces, was not detected. To improve the sensitivity of this assay, reverse transcriptase-PCR was used to generate probes from RNA extracted from fecal cultures. Coupled with a chemiluminescence detection method, this approach lowered the detection limit for E. coli from $\~1$ $\times 10$^6$ to ${\leq}$ 1 $\times 10$^5$ These results indicate that the membrane-array method with reverse transcriptase-PCR and chemiluminescence detection can simultaneously identify bacterial species present in fecal samples at cell concentrations as low as${\leq}$ 1 $\times 10$^5$ CFU per gram.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.2
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• pp.259-267
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• 1998

Typical characteristics of Mejus must be understood to get the basic data for setting up mass production system of traditional fermented soybean products. One hundred and twenty one Mejus were collected from various places and analyed. Most of shapes were rectangular and some were spherical, conical, cylindrical and doughnut types. The weight of Mejus was 0.4~4.2kg. Chemical analysis showed: moisture content, 9.73~58.22% ; pH, 4.95~8.15; acidity, 0.6~3.8% ; soluble protein content, 4.45~12.31%; soluble sugar content, 0.82~10.95%. Enzyme assay showed: $\alpha$-amylase activity, 5.0~874.2 units/g; $\beta$-amylase activity, 0.02~27.74units/g; acidic protease activity, 31.3~225.1unts/g; lipase activity 1.0~53.0units/g. Total viable cells were 3.72$\times$107~1.35$\times$1010cfu/g, and yeast and mold count 6.46$\times$104~8.91$\times$106cfu/g. respectively. $\alpha$-Amylase activity of a traditional Meju from Incheon showed the highest activity of 732.8 units/g(interior section) and 823.2units/g (exterior section). $\beta$-Amylase activity was the highest{3.57 units/g (interior sectin) and 4.25units/g (exterior section)} in Meju from Chunbuk. Acidic protease activity was the highest in sample from Seoul, whereas traditional Meju from Kyongnam showed the highest activity of 21.5units/g(interior section) and 37.5units/g(exterior sectin).

• Preventive Nutrition and Food Science
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• v.17 no.2
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• pp.172-176
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• 2012

Doenjang is a traditional Korean fermented soybean product that provides a major source of protein. In this study, a total of 18 different home-made doenjang samples were examined for the presence of foodborne pathogens and the total aflatoxin levels. Using an enzyme-linked immunosorbent assay to assess microbial quality and potential public health risk, we showed that total coliform levels in the doenjang samples ranged from 0 to $4.43{\pm}2.32{\times}10^6\;CFU/g$, and the maximum limit of Bacillus cereus was $4.67{\pm}2.0{\times}10^5\;CFU/g$. However, other foodborne pathogens, such as Staphylococcus aureus, Escherichia coli O157:H7 and Salmonella spp., were not detected among the tested samples. One of the samples (S3) showed a maximum limit of $42.2{\pm}9.1\;{\mu}g/kg$ for aflatoxin levels, which was above the safety limit allowed by the Codex Alimentarius Commission (CAC) regulatory agency. Further research is necessary to determine whether and how doenjang safety can be improved via elimination/reduction of microbial contamination during fermentation and storage or using microbial starter cultures for its fermentation.

• Journal of Veterinary Clinics
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• v.40 no.5
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• pp.323-329
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• 2023

Peripheral blood-derived mesenchymal stem cells (PB-MSCs) have shown promise in cell-based therapy, as they can be harvested with ease through minimally invasive procedures. This study aimed to isolate PB-MSCs from foals and mares and to compare the proliferation and cellular characteristics of the PB-MSCs between the two groups. Six pairs of mares and their foals were used in this study. MSCs were isolated from PB by direct plating in a tissue culture medium, and cell proliferation (population doubling time [PDT], and colony-forming unit-fibroblast assay [CFU-F]), and characterization (morphology, plastic adhesiveness, colony formation, trilineage differentiation) were examined. There was no significant difference in the PB-MSC yield, CFU-F, and PDT between the mares and foals. PB-MSCs from both mares and foals showed typical MSC characteristics in terms of spindle-shaped morphology, plastic adhesive properties, formation of colonies, trilineage differentiation. These results suggest that PB-MSCs isolated from horses, both adult horses, and foals, can be used for equine cell-based therapy.

• Molecules and Cells
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• v.39 no.11
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• pp.807-813
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• 2016

Escherichia coli are important indicator organisms, used routinely for the monitoring of water and food safety. For quick, sensitive and real-time detection of E. coli we developed a 2'F modified RNA aptamer Ec3, by Cell-SELEX. The 31 nucleotide truncated Ec3 demonstrated improved binding and low nano-molar affinity to E. coli. The aptamer developed by us out-performs the commercial antibody and aptamer used for E. coli detection. Ec3(31) aptamer based E. coli detection was done using three different detection formats and the assay sensitivities were determined. Conventional Ec3(31)-biotin-streptavidin magnetic separation could detect E. coli with a limit of detection of $1.3{\times}10^6CFU/ml$. Although, optical analytic technique, biolayer interferometry, did not improve the sensitivity of detection for whole cells, a very significant improvement in the detection was seen with the E. coli cell lysate ($5{\times}10^4CFU/ml$). Finally we developed Electrochemical Impedance Spectroscopy (EIS) gap capacitance biosensor that has detection limits of $2{\times}10^4CFU/mL$ of E. coli cells, without any labeling and signal amplification techniques. We believe that our developed method can step towards more complex and real sample application.

• Journal of Food Hygiene and Safety
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• v.38 no.5
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• pp.347-355
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• 2023

In this study, polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) were compared in terms of their ability to detect shiga-toxin-producing Escherichia coli (STEC). Various foods were artificially inoculated with STEC to evaluate the limit of detection (LOD), limit of quantification (LOQ), sensitivity, specificity, and efficiency of PCR and LAMP. The LODs were ≤10⁴ and ≤10³ CFU/mL for PCR and LAMP, respectively. The LOQs did not differ between PCR and LAMP. However, of the four considered food types, the sensitivities differed by a maximum of 11.1% for seasoned meat and by a minimum of 8.1% for ground beef. LAMP had higher sensitivity than that of PCR and 100% specificity for all four food types. Therefore, LAMP is a reliable molecular method for detecting STEC as comparable to PCR assay, and its specificity and sensitivity are superior to those of PCR, depending on the food type.

• Korean Journal of Ichthyology
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• v.29 no.1
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• pp.62-68
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• 2017

The Alaska pollock (Gadus chalcogrammus) belongs to the family Gadidae; it is a cold water fish, and has been developed as a novel aquaculture species in Korea. In this study, we describe ongoing surveillance for aquatic animal pathogens based on growth stages. We investigated bacterial flora in rearing water, and monitored pathogens; we also analyzed histopathological traits of abnormal fish. In rearing water, the total bacterial counts were $2.1{\times}10^3cfu/mL$ and Vibrio spp. (52%) were predominant in the larvae stage. In the juvenile and adult stages, the total bacterial counts were $3.4{\times}10^3$ and $3.2{\times}10^2cfu/mL$, respectively (with Pseudomonas sp. as the predominant species; 90% and 52%). This result revealed that the bacterial flora in rearing water changed depending on the feeding types. No virulent-bacteria or problematic viruses (VHSV, viral hemorrhagic septicemia virus; NNV, nervous necrosis virus; MBV, marine birnavirus) were detected from outwardly healthy fish using either culture or PCR assay. Some juveniles (less than 5%) had gas bubbles on the gill lamellae, degeneration of the corneal epithelium, and choroid gland degeneration, suggesting that these symptoms were caused by external injury and secondary infection by opportunistic bacteria. Disease management is important to cope with disease emergence in the novel aquaculture species Alaska pollock.