• Korean Journal Plant Pathology
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• v.10 no.2
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• pp.129-135
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• 1994

Ninety-two isolates of Fusarium species were obtained from Chinese corn samples. The predominant Fusarium species isolated from corn seeds were F. moniliforme, F. proliferatum, F. oxysporum and F. subglutinans, and all 13 species were identified. Each isolate was grown on autoclaved wheat grains and wheat cultures were fed by twenty-one-day-old female rats for the toxicity test. Twenty-six out of 92 isolates caused the death accompanying feed refusal, severe weight loss, liver damage, and hemorrhages in the stomach and intestines. Of the toxigenic isolates, 17 isolates of F. moniliforme, 4 of F. oxysporum, 3 of F. proliferatum, and one of each F. sporotrichioides and unknown species were lethal to rats. The analyses of fumonisin B1 production of the 26 toxigenic Fusarium isolates were carried out by thin layer chromatography and high-performance liquid chromatography, and fumonisin B1 was confirmed by mass spectrometry. Fumonisin B1 was produced in wheat culture at levels ranging from 280 $\mu\textrm{g}$/g to 3,952 $\mu\textrm{g}$/g by all of toxigenic F. moniliforme and F. proliferatum, but by none of the other toxigenic Fusarium species. The present results suggest the high possibility of natural occurrence of fumonisin B1 in corn samples imported from China.

• Korean Journal of Food Science and Technology
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• v.27 no.1
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• pp.119-123
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• 1995

In the present study, an attempt was made to investigate the effect of water activity(Aw) of rough rice on the growth of Fusarium moniliforme NRRL 13569 as well as on the production of Fumonisin $B_1(FB_1)$. The maximum growth of F. moniliforme and the production of $FB_1$, occurred at Aw 0.97 when the Aw of rough rice was controlled from 0.85 to 0.97. The fungal growth and $FB_1$, production decreased with the decrease of Aw. Sparse growth of fungus was observed even at Aw 0.85. $FB_1$ production on rough rice decreased considerably under Aw 0.97 and the trace amounts of $FB_1$ were observed at Aw 0.93 and 0.90. Therefore Aw required for the prevention of $FB_1$ production was turn out to be below 0.90 while that of fungal growth below 0.85.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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• pp.501-505
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• 1997

The effects of some nutrients and culture conditions on the growth and the production of fumonisin B$_{1}$, (FB$_{1}$) from Fusarium moniliforme NRRL 13569 were investigated in liquid culture. Xylose and soytone yielded the highest mycelial growth as the C- and N-source, respectively. The highest level of FB$_{1}$, was obtained when yeast extract was used as the N-source but no FB$_{1}$, from NaNO$_{3}$. While Fe$^{+++}$ showed inhibition effect on FB$_{1}$, production, Zn$^{++}$ enhanced the FB$_{1}$, production as well as the mycelial growth. FB$_{1}$, was maximally produced when the initial pH value and the specific surface area of the medium was adjusted to 5 and 1.4 cm$^{2}$/ml, respectively. FB$_{1}$ formation reached the maximum value (210, 000 ng/ml) in 30 days and then decreased in Czapek medium substitued with 1% xylose and 0.3% yeast extract, and supplemented with 0.2% NH$_{4}$H$_{2}$PO$_{4}$ where the initial pH value and the specific surface area of the medium are optimally controlled.

• Research in Plant Disease
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• v.18 no.4
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• pp.310-316
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• 2012

Gibberellea fujikuroi species (Gf) complex comprises at least 15 species, most of which not only causes serious plant diseases, but also produces mycotoxins including fumonisins. Here, we focused on the abilities of the field isolates belonging to the Gf complex associated with rice and corn, respectively in Korea to produce fumonisin, all of which were confirmed to carry FUM1, the polyketide synthase gene essential for fumonisin biosynthesis. A total of 88 Gf complex isolates (55 F. fujikuroi, 10 F. verticillioides, 20 F. proliferatum, 2 F. subglutinans, and 1 F. concentricum), and 4 isolates of F. commune, which is a non-member of Gf complex, were grown on rice substrate and determined for their production levels of fumonisins by a HPLC method. Most isolates of F. verticillioides and F. proliferatum, regardless of host origins, produced fumonisin $B_1$ and $B_2$ at diverse ranges of levels ($0.5-2,686.4{\mu}g/g$, and $0.7-1,497.6{\mu}g/g$, respectively). In contrast, all the isolates of F. fujikuroi and other Fusarium species examined produced no fumonisins or only trace amounts ($<10{\mu}g/g$) of fumonisins. Interestingly, the frequencies of relatively high fumonisin-producers among the F. proliferatum and F. fujikuroi isolates derived from corn were higher than those among the fungal isolates from rice. In addition, it is a first report demonstrating the ability of the FUM1-carrying F. commune isolates from rice to produce fumonisins.

• 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.15 no.4
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• pp.610-614
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• 2002

The purpose of this study was to investigate the prevalence of fumonisin contamination in corn and corn-based feeds in Taiwan. A total of 233 samples was collected from 8 feed mill factories located in four different regions in Taiwan. The presence of fumonisin $B_1$ ($FB_1$) and $B_2$ ($FB_2$) was determined by thin layer chromatograph, while the total fumonisin content was determined using immuno-affinity column cleanup and fluorometer quantitation. Our results showed that 55 samples of swine feeds had the highest percentage of incidence of $FB_1$ and $FB_2$ (41.8% and 41.8%, respectively), followed by 66 samples of duck feeds (40.9% and 37.8%). However, the percentage of incidence of $FB_1$ and $FB_2$ was much lower in 43 samples of broiler feeds (23.2% and 13.9%) and 69 samples of corn (17.3% and 10.1%). Corn and duck feeds were found to have a significant higher level of means of total fumonisins ($5.4{\pm}1.5$ and $5.8{\pm}0.6$ ppm, respectively) than swine feeds ($2.9{\pm}0.4$ ppm) and broiler feeds ($3.0{\pm}0.5$ ppm). Comparing fumonisins distribution in different regions, the highest percentage of $FB_1$ incidence (39.2%) was found in the eastern region of Taiwan, and total fumonisins level ($4.5{\pm}0.7$ ppm) was significantly higher than other regions. However, the highest percentage of $FB_2$ incidence (32.0%) was found in the central region of Taiwan. Trimonthly analysis of data showed that both high percentage of $FB_1$ and $FB_2$ incidence (39.3% and 37.7%) and total concentration of fumonisin ($5.7{\pm}0.4$ ppm) were found in the period of Jan. to Mar., The incidence and concentration were significantly higher than other trimothly periods. These results indicate that fumonisin B mycotoxins are both widespread and persistent in feed-grade corn and corn-based feeds in Taiwan.

• The Plant Pathology Journal
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• v.17 no.5
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• pp.281-289
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• 2001

We analyzed 88 strains of Gibberella fujikuroi (Analmorph: Fusarium section Liseola) from maize in Korea for mating population, mating type, fumonisin production vegetative compatibility, and random amplified polymorphic DNA (RAPD) patterns. We found 50 strains that were MATA-2, 22 that were MATA-1, 1 that was MATD-1, and 15 that were not reproducibly fertile with any of the mating type testers. Of the 50 MATA-2, 15 were female fertile, while 10 of the 22 MATA-1 strains were female fertile. A total of 1,138 nitrate non-utilizing (nit) mutants were recovered from a total of 88 strains. These strains were grouped into 39 vegetative compatability groups (VCGs) by demonstrating heterokaryosis between nit mutants. A single maize ear could be infected by more than one VCG of F. moniliforme. RAPD analysis measured genetic diversity among 63 strains of F. moniliforme. Several VCGs were distinguished by RAPD fingerprinting patterns. Most strains produced significant levels of fumonisins. However, 6 MATA-2 strains from a single VCG produced higher levels of fumonisin $\textrm{B}_3$ than that of fumonisin $\textrm{B}_1$ or $\textrm{B}_2$. From these data, we concluded that most Korean strains of F. moniliforme associated with maize belonged to mating population A and produced significant levels of fumonisins. Futhermore, RAPD analysis could differentiate strains associated with different VCGs.

• The Plant Pathology Journal
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• v.32 no.3
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• pp.173-181
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• 2016

Gene disruption by homologous recombination is widely used to investigate and analyze the function of genes in Fusarium fujikuroi, a fungus that causes bakanae disease and root rot symptoms in rice. To generate gene deletion constructs, the use of conventional cloning methods, which rely on restriction enzymes and ligases, has had limited success due to a lack of unique restriction enzyme sites. Although strategies that avoid the use of restriction enzymes have been employed to overcome this issue, these methods require complicated PCR steps or are frequently inefficient. Here, we introduce a cloning system that utilizes multi-fragment assembly by In-Fusion to generate a gene disruption construct. This method utilizes DNA fragment fusion and requires only one PCR step and one reaction for construction. Using this strategy, a gene disruption construct for Fusarium cyclin C1 (FCC1), which is associated with fumonisin B1 bio-synthesis, was successfully created and used for fungal transformation. In vivo and in vitro experiments using confirmed fcc1 mutants suggest that fumonisin production is closely related to disease symptoms exhibited by F. fujikuroi strain B14. Taken together, this multi-fragment assembly method represents a simpler and a more convenient process for targeted gene disruption in fungi.

• Microbiology and Biotechnology Letters
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• v.24 no.1
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• pp.119-125
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• 1996

In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishment of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin $B_1(FB_1)$ conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit was tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1:16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigated by the ciELISA, it was very low against $B_3$ (2%) but high against fumonisin $B_2$ (179%). The sensitivity of the ELISAs was also very high, because the detection limit for $FB_1$ was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of $FB_1$ was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of $FB_1$ from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

• 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.