• Proceedings of the Korean Vacuum Society Conference
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• 2014.02a
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• pp.126-126
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• 2014

Label-free biomolecular assay based localized surface plasmon resonance (LSPR) of noble metal nanoparticles enables simple and rapid detection with the use of simple equipment. Nanosized metal nanoparticles exhibit a strong absorption band when the incident light frequency is resonant with the collective oscillation of the electrons, which is known as the LSPR. Here we demonstrate localized surface plasmon resonance (LSPR) substrates such as plasmonic Au nanodisks fabricated by a nanoimprinting process and gold nanorod-immobilized surfaces and their applications to highly sensitive and/or label-free biosensing. To increase detection sensitivity various bioreceptors weree designed. A single chain variable fragment (scFv) was used as a receptor to bind C-reactive protein (CRP). The results of this effort showed that CRP in human serum could be quantitatively detected lower than 1 ng/ml. Aptamers, which were immobilized on gold nanorods, were used to detect mycotoxins. The specific binding of ochratoxin A (OTA) to the aptamer was monitored by the longitudinal wavelength shift of LSPR peak in the UV-Vis spectra resulting from the changes of local refractive index near the GNR surface induced by accumulation of OTA and G-quadruplex structure formation of the aptamer. According to our results, OTA could be quantitatively detected lower than 1 nM level. Additionally, aptamer-functionalized GNR substrate was quite robust and can be regenerated many times by rinsing at 70 OC to remove bound target. During seven times of washing steps, the developed OTA sensing system could be reusable. Moreover, the proposed biosensor exhibited selectivity over other mycotoxins with an excellent recovery for detection in grinded corn samples, suggesting that the proposed LSPR based aptasensor plays an important role in label-free detection of mycotoxins.

• Korean Journal of Veterinary Service
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• v.42 no.3
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• pp.127-133
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• 2019

The purpose of this study was to investigate the contamination level of representative mycotoxins that have adverse effects on livestock by using LC-MS/MS method and to utilize the results as basic data for the establishment of quality control system for feed, and to provide information on production and storage. A total of nine mycotoxins, including aflatoxin $B_1$, aflatoxin $B_2$, aflatoxin $G_1$, aflatoxin $G_2$, ochratoxin A, fumonisin $B_1$, fumonisin $B_2$, deoxynivalenol (DON), zearalenone (ZEN) were simultaneously analyzed in LC-MS/MS under ESI positive mode. Fumonisin $B_1$ and fumonisin $B_2$ were detected from 3 cases of 75 forage produced in Incheon area, the detection rate was 4.0%. The detection concentration was 0.01~0.02 mg/kg, which was lower than the domestic recommended limit. Fumonisins were detected in a slightly different manner from the results of mycotoxin studies reported in Korea, which is attributed to the high temperature and dry summer weather of the year. The result of LC-MS/MS method performance of 9 mycotoxins, the recovery of DON was quite low as $41.53{\pm}3.91%$ that is not suitable for simultaneous analysis. This is probably due to that the extract solution used in this study was not suitable for the extraction of DON, along with the characteristics of a very dry forage. For the study of mycotoxins in Incheon area forage for the first time, further investigation is needed for the safe supply of livestock products.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.12
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• pp.1903-1912
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• 2018

Objective: To evaluate the effect of lactic acid bacteria and storage temperature on the microbial, chemical and mycotoxin composition of corn silage. Methods: Corn was harvested at 32.8% dry matter, and chopped to 1 to 2 cm. The chopped material was subjected to three treatments: i) control (distilled water); ii) $1{\times}10^6$ colony forming units (cfu)/g of Lactobacillus plantarum; iii) $1{\times}10^6cfu/g$ of Pediococcus pentosaceus. Treatments in triplicate were ensiled for 55 d at $20^{\circ}C$, $28^{\circ}C$, and $37^{\circ}C$ in 1-L polythene jars following packing to a density of approximately $800kg/m^3$ of fresh matter, respectively. At silo opening, microbial populations, fermentation characteristics, nutritive value and mycotoxins of corn silage were determined. Results: L. plantarum significantly increased yeast number, water soluble carbohydrates, nitrate and deoxynivalenol content, and significantly decreased the ammonia N value in corn silage compared with the control (p<0.05). P. pentosaceus significantly increased lactic acid bacteria and yeast number and content of deoxynivalenol, nivalenol, T-2 toxin and zearalenone, while decreasing mold population and content of nitrate and 3-acetyl-deoxynivalneol in corn silage when stored at $20^{\circ}C$ compared to the control (p<0.05). Storage temperature had a significant effect on deoxynivalenol, nivalenol, ochratoxin A, and zearalenone level in corn silage (p<0.05). Conclusion: Lactobacillus plantarum and Pediococcus pentosaceus did not decrease the contents of mycotoxins or nitrate in corn silage stored at three temperatures.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.1
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• pp.21-29
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• 2013

The studies were conducted to investigate real condition of mycotoxin contamination in the fields before harvest and by the storages of rice straw used as roughage in Korea. It was found mycotoxin contamination before harvest of rice straw that the rice plants were detected deoxynivalenol at the initial growth, ochratoxin A and deoxynivalenol at the middle growth, and deoxynivalenol and zearalenone at the harvest periods in the fields. Also, the rice plants were contaminated by various fungi such as Fusarium sp., Fusarium proliferatum, Penicillium sp., Gibberella sp., Gibberella zeae, Mucor circinelloides and Aspergillus oryzae. The levels of fungal contamination were $10^{3-4}$ cfu/g at the initial growth, and $10^{4-5}$ cfu/g at the middle growth and harvest periods. All storage types of rice straw were contaminated with zearalenone, deoxynivalenol and ochratoxins A. The samples of rice straw contaminating mycotoxins were account for 3% in bundle rice straw, and 38% in both types of square rice straw and rice round bale silage, respectively. When 105 samples of rice bale silage were analyzed for mycotoxins depending on the regional area, mycotoxin contamination was found in 46% of total samples in Korea. Regional contaminations of mycotoxins were respectively 48, 33, 40, 50 and 57% of samples in Gyeonggi, Gangwon, Chungcheng, Yeongnam and Honam area. Rice round bale silage was contaminated by three kinds of mycotoxins (zearalenone, deoxynivalenol and ochratoxinsA) in the all of area without Chungcheong area where was contaminated zearalenone and deoxynivalenol. Ochratoxins A, deoxynivalenol and zearalenone were respectively determinated with the average levels of 2.6, 413 and $338{\mu}g/kg$ in rice round bale silage for the overall area, even if it was some difference depending on each regional area. Therefore, the above results clearly show that the rice straws were exposed to the contamination by mycotoxin and mycotoxigenic fungi before harvest in the fields, and mycotoxin contamination was not dependent on the regional area or the storage types such as bundle rice straw, square rice straw and rice round bale silage.

• Journal of Food Hygiene and Safety
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• v.36 no.1
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• pp.24-33
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• 2021

For this study, we surveyed concentrations of 8 mycotoxins (aflatoxin B1, B2, G1, G2, ochratoxin A, fumonisin B1, B2 and zearalenone) in agricultural products used for food and medicine by liquid chromatography-tandem mass spectrometry and conducted a risk assessment. Samples were collected at the Yangnyeong Market in Seoul, Korea, between January and November 2019. Mycotoxins were extracted from these samples by adding 0.1% formic acid in 50% acetonitrile and cleaned up by using an ISOLUTE Myco cartridge. The method was validated by assessing its matrix effects, linearity, limit of detection (LOD), limit of quantification (LOQ), recovery and precision using four representative matrices. Matrix-matched standard calibration was used for quantification and the calibration curves of all analytes showed good linearity (r2>0.9999). LODs and LOQs were in the range of 0.02-0.11 ㎍/kg and 0.06-0.26 ㎍/kg, respectively. Sample recoveries were from 81.2 to 118.7% and relative standard deviations lower than 8.90%. The method developed in this study was applied to analyze a total of 187 samples, and aflatoxin B1 was detected at the range of 1.18-7.29 ㎍/kg (below the maximum allowable limit set by the Ministry of Food and Drug Safety, MFDS), whereas aflatoxin B2, G1 and G2 were not detected. Mycotoxins that are not regulated presently in Korea were also detected: fumonisin (0.84-14.25 ㎍/kg), ochratoxin A (0.76-17.42 ㎍/kg), and zearalenone (1.73-15.96 ㎍/kg). Risk assessment was evaluated by using estimated daily intake (EDI) and specific guideline values. These results indicate that the overall exposure level of Koreans to mycotoxins due to the intake of agricultural products used for food and medicine is unlikely to be a major risk factor for their health.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.4
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• pp.451-462
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• 2011

The purpose of this study was to investigate fungi and mycotoxin contamination of the rice straw bale silage in Korean. It was tested the 33 samples of rice straw round bale silage with various condition which fed cattle in the farm. The level of fungal contamination was $2.1{\times}10^6\;cfu\;g^{-1}$ in the average and $9.2{\times}10^8\;cfu\;g^{-1}$ in the maximum. The fungal contamination was detected in the all of normal samples which good condition of rice straw bale silage. When the fungi was isolate and identify, it was found 28 species and mycotoxin producing fungi were 8 species as following as Aspergillus flavus, Aspergillus fumigatus, Fusarium culmorum, Fusarium verticillioides, Penicillium carneum, Penicillium paneum, Penicillium roqueforti, Penicillium viridicatum. Specially, Penicillium paneum was found 42% of samples and Aspergillus sp. (A. flavus, A. fumigatus) are 21% of samples. In case of mycotoxin contamination, the 42% of samples are detected more than one kind of mycotoxin. Some samples are contaminated three kinds of mycotoxin. This study was not found aflatoxin ($B_1$, $B_2$, $G_1$, $G_2$) and fumonisin ($B_1$, $B_2$), but were detected the contamination of ochratoxin A (1.0~5.8 ug/kg), deoxynivalenol (DON, 156.0~776.7 ug/kg) and zearalenone (ZON, 38.0~750.0 ug/kg). Therefore, the above results show that rice straw round bale silage expose on hazard factors as mycotoxigenic fungi and mycotoxin contamination, and than need more research about mycotoxin in animal feed to protect animal and human healthy.

• Culinary science and hospitality research
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• v.23 no.8
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• pp.153-162
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• 2017

Bacillus coagulans was added as probiotics in instant coffee with microground roasted coffee which is recently rising premium coffee and the instant coffee (ProBio coffee) was compared with 3 commercial instant coffees with microground roasted coffee in quality characteristics to understand the competitiveness of ProBio coffee. In sensory evaluation, ProBio coffee had inferior aroma intensity and overall acceptance compared with control group (brewed coffee) (p<0.05) but it had equal quality or more compared with 3 commercials. Total polyphenol content, chlorogenic acid content, DPPH free radical scavenging activity and caffeine content were $110.72{\pm}1.99mg/g$, $2,700{\pm}20mg/g$, $146.22{\pm}3.62TEAC\;mg/g$, $28.1{\pm}3.2ppm$ respectively. In general quality characteristics, water content, solubility, particle size and particle strength of ProBio coffee were similar to general instant coffee but had darker color than it. In safety assessment, acrylamide content was $502{\pm}10ppb$, and residual pesticides and ochratoxin A were not detected. Over 70% of B. coagulans were maintained in 4 months of storage and 16 months of shelf life was predicted in ProBio coffee by $Q_{10}$ model. Therefore, ProBio coffee was confirmed to have a sufficient product competiveness compared with 3 commercials.

• Korean Journal of Poultry Science
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• v.43 no.2
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• pp.111-118
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• 2016

Mycotoxins are secondary metabolites produced by molds, such as Aspergillus, Fusarium and Penicillium, that have adverse effects on animals and humans. Aflatoxin, ochratoxin, zearalenone, fumonisin and deoxynivalenol are the mycotoxins of greatest agro-economic importance and cause acute disease called mycotoxicoses. Mycotoxicosis in poultry birds results in decreased meat/egg production, immunosuppressant, and hepatotoxicosis. Some of toxins or their metabolites may be retained in animal or human tissues and induce health problems. This study was designed to develop a sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS) method for the simultaneous detection and quantification of mycotoxins, such as aflatoxin $B_1$, aflatoxin $M_1$, ochratoxin A, zearalenone, fumonisin B and deoxynivalenol, in chicken liver and kidney tissues. The mycotoxins were extracted and purified using modified QUECHERS methods, separated by LC and detected by an electrospray ionisation interface (ESI) and tandem MS. Good precision and linearity were observed for most of six mycotoxins. The recovery test for each mycotoxin in liver and kidney tissues mostly indicated good average recovery rates between 80.94% and 98.10% and the coefficient of variation mostly under 13.78%, except for aflatoxin $M_1$ and fumonisin $B_1$. The limit of detection (LOD) for six mycotoxins was $7.6{\sim}145.79{\mu}g/kg$ in liver tissues and $6.07{\sim}197.20{\mu}g/kg$ in kidney tissues. The quantification limits (LOQ) for 6 mycotoxins were in the range $23.04{\sim}441.78{\mu}g/kg$ in liver tissues and $18.40{\sim}597.59{\mu}g/kg$ in kidney tissues, respectively. The developed multi-mycotoxin method in this study permits simultaneous, simple, and rapid determination of several co-existing mycotoxins in chicken liver and kidney tissues.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1197-1204
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• 2018

Objective: The objective was to characterize the fermentative and microbiological profile of Tifton 85 bermudagrass haylage with different layers of polyethylene film and storage time. Methods: The experimental design consisted of a randomized block design with four and six wrapping layers (100 and 150 microns in total. respectively) allocated in the main plots, through repeated measures analysis (30, 60, and 90 days of storage) with four replicates. Results: The storage time and number of wrapping layers did not show changes in the population of Clostridium and lactic acid bacteria. A decrease was observed in the enterobacteria population with an increase in the storage period in the two wrapping layers studied. Upon opening of the haylage at 30 days, the population of Bacillus was lower in haylages made with six layers of wrapping (3.63 log colony forming units/g). No growth of Listeria sp. or Salmonella sp. was observed during the experimental period. The fungal genera with a greater occurrence were Penicillium sp. and Fusarium sp. The following mycotoxins were not detected: ochratoxin A, fumonisins, and zearalenone. Relative to the organic butyric, propionic, and acetic acids, the haylages presented a low concentration of lactic acid; this may have prevented a drop in the pH, which was high when the silos were opened (5.4). The levels of ammoniacal nitrogen and soluble carbohydrates presented no variation among the number of wrapping layers, with an overall average of 35.55 and 38.04 g/kg. Conclusion: Tifton 85 bermudagrass haylage wrapped with four and six layers presented adequate fermentation and microbiological characteristics in the evaluated periods.

• Journal of Food Hygiene and Safety
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• v.34 no.3
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• pp.269-276
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• 2019

A colorimetric rapid detection method based on acetylcholinesterase (AChE) was developed for the analysis of organophosphorus (OP) and carbamate (CB) pesticides. The AChE catalyzes acetylthiocholine into thiocholine having (-) and (+) charges, and the (+) charge results in gold nanoparticle (GNP) aggregation. The in-activation of AChE by OP and CB has been well known. In order to optimize the colorimetric method, optimal dilution times of commercial serum containing AChE, diameter of GNP, and concentration of acetylthiocholine were tested as a key parameter. The colorimetric detection limits of the method were 7.5 ng/mL for both dimethyl amine and carbofuran pesticides in 60% ethanol. No cross-reaction to other chemicals, such as aflatoxin B1 and ochratoxin A, which can be contaminated with pesticides in agricultural products, was observed. Recoveries from lettuce, sesame leaf, and cabbage lettuce spiked with known concentrations of dimethyl amine and carbofuran were found to be ranged from 83.85 to 133.16%. These results indicated that the colorimetric rapid method based on AChE can be a useful tool for the sensitive, specific, rapid, and accurate detection of OP and CB pesticides in fresh vegetables.