• Tuberculosis and Respiratory Diseases
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• v.53 no.5
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• pp.497-509
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• 2002

Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.

• Food Science and Biotechnology
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• v.16 no.3
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• pp.337-342
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• 2007

Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Biosensors have shown great potential for the rapid detection of foodborne pathogens. Fiber-optic biosensors have been used to rapidly detect pathogens because they can be very sensitive and are simple to operate. However, many fiber-optic biosensors rely on manual sensor handling and the sandwich assay, which require more effort and are less sensitive. To increase the simplicity of operation and detection sensitivity, a binding inhibition assay method for detecting Listeria monocytogenes in food samples was developed using an automated, fiber-optic-based immunosensor: RAPTOR (Research International, Monroe, WA, USA). For the assay, fiber-optic biosensors were developed by the immobilization of Listeria antibodies on polystyrene fiber waveguides through a biotin-avidin reaction. Developed fiber-optic biosensors were incorporated into the RAPTOR to evaluate the detection of L. monocytogenes in frankfurter samples. The binding inhibition method combined with RAPTOR was sensitive enough to detect L. monocytogenes ($5.4{\times}10^7\;CFU/mL$) in a frankfurter sample.

• Journal of Microbiology and Biotechnology
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• v.24 no.3
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• pp.427-430
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• 2014

We developed a novel immunochromatographic assay (ICA) (EZ-Step VanA rapid kit; Dinona, Korea) for the detection of VanA ligase from vancomycin-resistant enterococci (VRE). Of eight monoclonal antibodies screened by ELISAs, the VanA ligase ICA constructed with 1H9 plus 3G11 showed the greatest reactivity. The detection limit of the kit was $6.3{\times}10^6$ CFU per test. Of 127 vancomycin-resistant microorganisms, 100 vanA VRE were positive in the VanA ligase ICA, and 27 non-vanA vancomycin-resistant isolates were negative. These results were consistent with those of the PCR analyses. Thus, our ICA is a reliable and easy-to-use immunological assay for detecting VanA-producing VRE in clinical laboratories.

• Food Science and Biotechnology
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• v.16 no.4
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• pp.515-519
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• 2007

Rapid immunochromatographic assay (ICA) kits were developed using flagella-specific monoclonal antibodies (MAbs) and rabbit polyclonal antibodies for screening Listeria spp. in food. The establishment of different formats, MAb 2B1 as capture antibody and MAb 7A3 or rabbit polyclonal antibodies as detector antibody, was compared. The 2 formats of the ICA kit were shown to have specific reactions with Listeria and no cross-reactivity with any of the non-Listeria including Escherichia coli O157:H7 and Salmonella enteritidis. The detection limits of the ICA kit using the combination of gold-labeled MAb 7A3 and MAb 2B1 showed $1{\times}10^5$ and $1{\times}10^6\;CFU/0.1\;mL$ at 22 and $30^{\circ}C$, respectively. The other format of the ICA kit using the combination of gold-labeled rabbit polyclonal antibodies and MAb 2B1 showed $1{\times}10^6\;CFU/0.1\;mL$ at $22^{\circ}C$ but weak signal at 30 culture. The format utilizing MAb was more sensitive than the one using polyclonal antibodies for capture antibody. Samples contaminated with L. monocytogenes 4b culture (9-10, 5-6, and 1-2 CFU/mL) on pork and pasteurized milk were confirmed as positive results. Current data suggests that this ICA kit is a rapid, simple and effective tool to screen for Listeria spp. in food.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.9
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• pp.1415-1422
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• 2014

Hot water extraction concentrate was prepared from Alliun hookeri root (AHR) to evaluate its applicability to yogurt. The highest antioxidant activity of hot water concentrates was obtained under extraction conditions of 4 hr at $95^{\circ}C$. Antioxidant activities measured by DPPH radical assay, ABTS radical cation assay, reducing power, and cheating activity were highly correlated with total phenolic (89.51 mg/g) and total flavonoid (52.71 mg/g) contents, with R values of 0.94 and 0.96, respectively. Yogurt was fermented with a commercial lactic acid bacteria mixed strain (Yo-mix$^{TM}$ 305) for 10 hr at $42^{\circ}C$ after addition of 0~10% (w/w) hot water concentrates from AHR to yogurt base. As fermentation proceeded, pH and $^{\circ}Brix$ of yogurt decreased from 6.57~6.60 to 4.34~4.51 and from 8.10~8.90% to 4.60~5.25%, respectively, whereas titrate acidity, viscosity, and viable cell numbers increased from 0.22~0.23% to 1.01~1.10%, from $0mPa{\cdot}s$ to $202.55{\sim}290.50mPa{\cdot}s$, and from 6.40~6.80 log CFU/mL to 8.60~9.20 log CFU/mL, respectively. There was no significant difference in any sensory attribute between the control and 2.5% addition group, suggesting that 2.5% hot water concentrate from AHR could be used to manufacture yogurt.

• Preventive Nutrition and Food Science
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• v.19 no.2
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• pp.115-123
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• 2014

In the current study, wheat flour was mixed with high quality cassava flour (HQCF) in several ratios: 90:10, 80:20, 70:30, and 60:40, and used to prepare 10%, 20%, 30%, and 40% National Root Crops Research Institute (NRCRI) cassava bread, respectively. 100% wheat bread was prepared as a control (100% wheat bread). Five bread samples were prepared per group. Antioxidant assays [i.e., 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) scavenging assay, reducing power assay] revealed that the bread samples had considerable antioxidant capacities. Substitution of wheat flour with HQCF at various concentrations resulted in dose dependent decreases in the mineral and protein contents of the resulting bread samples. The crude fiber content of the bread samples was minimal, while the carbohydrate content of the bread samples ranged from 43.86% to 48.64%. A 20% substitution of wheat flour with HQCF yielded bread samples with a general acceptability that was comparable to that of 100% wheat bread. The mean bacteria counts of the bread samples ranged from $2.0{\times}10^3CFU/mL$ to $1.4{\times}10^4CFU/mL$, while the fungal counts ranged from 0 CFU/mL to $3{\times}10^3CFU/mL$. There was a positive correlation between the DPPH antioxidant activities and the reducing powers of the bread samples ($R^2=0.871$) and a positive correlation between the DPPH antioxidant activities and the flavonoid contents of the bread samples ($R^2=0.487$). The higher microbial load of the NRCRI cassava bread samples indicates that these bread samples may have a shorter shelf life than the 100% wheat bread. The significant positive correlation between total flavonoid content and reducing power ($R^2=0.750$) suggests that the flavonoids present in the lipophilic fractions of the bread samples could be responsible for the reductive capacities of the bread samples.

• Food Science of Animal Resources
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• v.44 no.2
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• pp.372-389
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• 2024

This study investigated the efficacy of ultraviolet-C (UV-C) irradiation in enhancing the quality of raw bovine milk by targeting microbial populations and lipid peroxidation, both of which are key factors in milk spoilage. We categorized the raw milk samples into three groups based on initial bacterial load: low (<3 Log 10 CFU/mL), medium (3-4 Log 10 CFU/mL), and high (>4 Log 10 CFU/mL). Using a 144 W thin-film UV-C reactor, we treated the milk with a flow rate of 3 L/min. We measured the bacterial count including standard plate count, coliform count, coagulase-negative staphylococci count, and lactic acid bacteria count and lipid peroxidation (via thiobarbituric acid reactive substances assay) pre- and post-treatment. Our results show that UV-C treatment significantly reduced bacterial counts, with the most notable reductions observed in high and medium initial load samples (>4 and 3-4 Log 10 CFU/mL, respectively). The treatment was particularly effective against coliforms, showing higher reduction efficiency compared to coagulase-negative staphylococci and lactic acid bacteria. Notably, lipid peroxidation in UV-C treated milk was significantly lower than in pasteurized or untreated milk, even after 72 hours. These findings demonstrate the potential of UV-C irradiation as a pre-treatment method for raw milk, offering substantial reduction in microbial content and prevention of lipid peroxidation, thereby enhancing milk quality.

• Food Science of Animal Resources
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• v.30 no.4
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• pp.590-596
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• 2010

Meat and meat products are a potential source of food-borne pathogens, including Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7, and Bacillus cereus. A sensitive and specific PCR assay for the detection of these pathogens in meat and meat products was developed in this study, as part of a broader effort to reduce the potential health hazards posed by these pathogens. Initially, PCR conditions were standardized with purified DNA. Under standard conditions, the detection level for PCR was as low as 10 pg of purified bacterial DNA. After overnight growth of bacteria in a broth medium, as few as $10^2$ CFU of bacteria were detected by PCR assay. The primers employed in the PCR assay were found to be highly specific for individual organisms, and evidenced no cross-reactivity with heterologous organisms. Additionally, the multiplex PCR assays also amplified some target genes from the four pathogens, and multiplex amplification was obtained from as little as 10 pg of DNA, thus illustrating the excellent specificity and high sensitivity of the assay. In conclusion, this PCR-based technique provides a sensitive and specific method for the detection of S. aureus, Salmonella spp., E. coli O157:H7, and B. cereus in meat and meat products.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1855-1862
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• 2016

Cronobacter sakazakii (C. sakazakii) is a foodborne pathogen, posing a high risk of disease to infants and immunocompromised individuals. In order to develop a quick, easy, and sensitive assay for detecting C. sakazakii, a rabbit anti-C. sakazakii immunoglobulin G (IgG) was developed using sonicated cell protein from C. sakazakii. The developed anti-C. sakazakii (IgG) was of good quality and purity, as well as species-specific. The developed rabbit anti-C. sakazakii IgG was attached to the surface of a sulforhodamine B-encapsulated liposome to form an immunoliposome. A test strip was then prepared by coating goat anti-rabbit IgG onto the control line and rabbit anti-C. sakazakii IgG onto the test line, respectively, of a plastic-backed nitrocellulose membrane. A purple color signal both on the test line and the control line indicated the presence of C. sakazakii in the sample, whereas purple color only on the control line indicated the absence of C. sakazakii in the sample. This immunochromatographic strip assay could produce results in 15 min with a limit of detection of $10^7CFU/ml$ in C. sakazakii culture. The immunochromatographic strip assay also showed very good specificity without cross-reactivity with other tested Cronobacter species. Based on these results, the developed immunochromatographic strip assay is efficient for the detection of C. sakazakii and has high potential for on-site detection.

• Korean Journal of Soil Science and Fertilizer
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• v.46 no.6
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• pp.615-622
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• 2013

The purpose of this study was to evaluate microbial contamination of melons in Korea. A total of 123 samples including melon fruits, leaves, seeds, soils, and irrigation water were collected from farms and markets to detect total aerobic bacteria, coliform, Escherichia coli, and pathogenic bacteria such as Bacillus cereus, Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus. Samples were collected from Iksan and Nonsan farms to monitor bacterial levels on pre-market melons. The total aerobic and coliform bacteria on melon cultivation were between 0.43 and 6.65 log CFU $g^{-1}$, and 0.67 and 2.91 log CFU $g^{-1}$, respectively. Bacillus cereus, a fecal coliform, was detected in soils and melon leaves from Iksan farm at 2.95, 0.73 log CFU $g^{-1}$, respectively, and in soils from Nonsan farm at 3.16 log CFU $g^{-1}$. Market melon samples were collected to assay bacterial load on melon being sold to consumers. The contamination levels of total aerobic bacteria in agricultural markets, big-box retailers, and traditional markets were 4.82, 3.94, 3.99 log CFU $g^{-1}$, respectively. The numbers of coliform in melon on the markets ranged from 0.09 to 0.49 log CFU $g^{-1}$. Listeria monocytogenes, Salmonella spp., and Staphylococcus aureus were not detected in any samples. The count of total aerobic bacteria on melon seeds ranged from 0.33 to 3.34 log CFU $g^{-1}$. This study found that irrigation water, soil, manure and various farm work activities including post-harvest processes were latent sources of microbial contamination. These results suggest that hygienic management and monitoring of soil, water, and agricultural material should be performed to reduce microbial contamination in melon production.