• Food Science and Biotechnology
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• v.18 no.3
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• pp.641-648
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• 2009

A one-step simultaneous immunchromatographic (OS-ICG) assay using colloidal gold-monoclonal antibody (gold-MAb) conjugates was developed for the rapid multianalysis of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in feed samples. Visual detection limits for AFB1 and OTA were 0.5 and 2.5 ng/mL, respectively, and the results were obtained within 15 min. Matrix interference from the feed extracts was efficiently reduced by appropriate dilution with buffer. Cut-off values of the OS-ICG assay for the feed spiked with AFB1/OTA mixtures (5/5, 10/10, 25/25, 50/55, 100/100 ${\mu}g/kg$) were 10 and 50 ${\mu}g/kg$ for AFB1 and OTA. The comparative analyses of 65 feed samples by OS-ICG, enzyme-linked immunosorbent assay (ELISA), and high performance liquid chromatography (HPLC) showed good agreement. In this study, we confirmed that simultaneous analysis based on immunoassay is possible and it can be used as an on-site multianalysis of AFB1 and OTA in feed, food, and agricultural products.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.2
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• pp.198-205
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• 2011

This study was conducted to investigate the effect of feed types on Ochratoxin A (OTA) degradation by Korean native goats. Rumen fluid from canulated goats fed whole roughage or 50% roughage served as a source of micro-organisms. Experiments were undertaken i) to investigate OTA degradation ability in a $2{\times}4$ factorial arrangement with different feed types (100% roughage vs. 50% roughage) and rumen fluid fractions (whole rumen fluid, cells, autoclaved rumen fluid and supernatant) supplemented with OTA ii) to evaluate OTA degradation by the rumen fluid of goats fed two different diets at different time points (0, 3, 6, 9 and 12 h) of feeding iii) to isolate potential rumen microorganisms and iv) to identify elements responsible for OTA degradation. Rumen fluid from goats fed 100% roughage had higher (p<0.05) OTA degradability than 50% roughage diets. OTA degradation based on rumen fluid collection times showed that rumen fluid at 0 h showed significantly higher (p<0.05) degradability. Carboxypeptidase A (CPA) enzyme has been reported to be responsible for OTA degradation. Thus, using real time PCR, primers designed to target the CPA gene from Bacillus licheniformis could be amplified using genomic DNA from rumen fluid of goats and sequenced, thus enabling evaluation of the Bacillus population under different feeding condition and times. Our findings showed that the Bacillus population was significantly higher (p<0.05) before feeding (0 h) in animals which were fed a whole roughage diet, giving indirect evidence of OTA degradation being influenced by Bacillus sps. Thus, it can be concluded that OTA degradability is influenced by feed, feeding time and Bacillus licheniformis population.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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• pp.505-513
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• 2018

Objective: This experiment investigated the effects of aflatoxin B1 (AFB1) alone or mixed with ochratoxin A (OTA) and/or zearalenone (ZEA) on the metabolism, immune function, and antioxidant status of dairy goats. Methods: Fifty lactating Laoshan dairy goats were randomly assigned to one of five treatment groups (n = 10) for 14 days. Goats were fed no additive (control) or administered with $50{\mu}g\;AFB1/kg$ dry matter (DM) (AFB1), $50{\mu}g\;AFB1/kg$ $DM+100{\mu}g\;OTA/kg$ DM (AFB1+OTA), $50{\mu}g\;AFB1/kg$ $DM+500{\mu}g\;ZEA/kg$ DM (AFB1+ZEA), or $50{\mu}g\;AFB1/kg$ $DM+100{\mu}g\;OTA/kg$ $DM+500{\mu}g\;ZEA/kg$ DM (AFB1+OTA+ZEA). Results: Dry matter intake and milk production were lower in goats fed AFB1+OTA+ZEA than in controls. Supplementation with AFB1, OTA, and ZEA significantly decreased red blood cell count, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, and mean platelet volume, and significantly increased white blood cell count, when compared with the control group. Compared with control, the combination of AFB1, OTA, and ZEA significantly increased alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities, total bilirubin (TBIL), interleukin-6, and malondialdehyde (MDA), but significantly reduced immunoglobulin A concentration, the activities of superoxide dismutase (SOD) and glutathione peroxides (GSH-Px), and total antioxidant capacity (T-AOC) in serum. Administration of AFB1 combined with OTA led to higher ALP, ALT, TBIL, and MDA, as well as lower milk production, SOD and GSH-Px activities, and T-AOC, than administration of AFB1 combined with ZEA. Conclusion: The mixture of AFB1, OTA, and ZEA exerted the greatest adverse effects on dairy goats, meanwhile the deleterious damage of the other mycotoxin combinations were in varying degrees. The findings of this study could provide guidance for the prevention and treatment of the consequences of contamination of animal feeds with combinations of mycotoxin.

• Journal of Food Hygiene and Safety
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• v.15 no.1
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• pp.41-45
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• 2000

Induction of DNA fragmentation of rat embryonic midbrain cells was studied to see whether apoptosis plays a role in OTA-induced microcephaly observed in cultured rat whole embryos during embryogenesis. We first cultured whole embryos (prepared from day 9.5 gestation rats) for 48 hrs with OTA and found that OTA induced microcephaly in cultured rat whole embryos. We also examined whether the microcephaly seen in cultured whole embryos is partially related to the increase of apoptosis of undifferentiated embryonic midbrain cells. Embryonic midbrain cells were prepared from day 12 gestation rat embryos, and cultured in the mixture media of Dulbecco's modified eagle's medium nutrient and Ham's F12 (1：1) containing 10% Nuserum, 100 $\mu\textrm{g}$/ml of streptomycin and 100 units/ml of penicillin for 96 hrs. Induction of DNA fragmentation was increased by 0.25-1 $\mu\textrm{g}$/ml OTA in a dose dependent manner in the embryonic midbrain cells. We also tested whether increase of apoptosis by OTA would be associated with change of apoptosis-related proteins (TNF-$\alpha$ and P$^{53}$ ) level in embryonic midbrain cells. OTA also increased TNF-$\alpha$ and P$^{53}$ levels. These results show that OTA induced microcephaly in cultured whole embryos and this effect may be at least a part due to the induction of apoptosis and apoptosis-related protein levels of undifferentiated embryonic midbrain cells.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.14
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• pp.5987-5991
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• 2015

Our previous study detected aflatoxins in red chili peppers from Chile, Bolivia, and Peru, each of which have a high incidence of gallbladder cancer (GBC). Since the aflatoxin B1 concentration was not so high in these peppers, it is important to clarify the presence of other mycotoxins. Here we attempted to determine any associations between the concentrations of aflatoxins and ochratoxin A (OTA) in red chili peppers, and the corresponding GBC incidences. We collected red chili peppers from three areas in Peru: Trujillo (a high GBC incidence area), Cusco (an intermediate GBC incidence area), and Lima (a low GBC incidence rate), and from Chile and Bolivia. Aflatoxins and OTA were extracted with organic solvents. The concentrations of aflatoxins B1, B2, G1, and G2, and OTA were measured by high-performance liquid chromatography. The values obtained were compared with the incidence of GBC in each area or country. All of the red chili peppers from the three areas showed contamination with aflatoxins below the Commission of the European Communities (EC) recommended limits ($5{\mu}g/kg$), but the OTA contamination of two samples was above the EC recommended limit ($15{\mu}g/kg$). The mean concentrations of OTA in the peppers from Chile (mean $355{\mu}g/kg$, range < $5-1,059{\mu}g/kg$) and Bolivia (mean $207{\mu}g/kg$, range $0.8-628{\mu}g/kg$), which has a high incidence of GBC, were higher than that in Peru ($14{\mu}g/kg$, range < $5-47{\mu}g/kg$), which has an intermediate GBC incidence. The OTA contamination in the red chili peppers from Chile, Bolivia, and Peru was stronger than that of aflatoxins. Our data suggest that OTA in red chili peppers may be associated with the development of GBC.

• Proceedings of the Korean Society of Toxicology Conference
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• 2001.05a
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• pp.168-168
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• 2001

The mycotoxin, ochratoxin A (OTA) has been known to induce microcephaly in animals and in vitro whole embryo. Cytotoxic effect and inhibition of cell differentiation were proposed as underlying mechanisms responsible for OTA-induced microcephaly.(omitted)

• Journal of Food Hygiene and Safety
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• v.24 no.4
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• pp.299-306
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• 2009

Aflatoxin (AF) and Ochratoxin A (OTA) are carcinogenic and possible carcinogenic mycotoxins respectively produced by Aspergillus spp or Penicillium spp. The study for contamination survey and proposal for reduction of mycotoxin in red pepper were carried out. 192 samples were collected at such various stages and markets as pre/post-harvest stages, internet shopping mall /super-market and small stakeholder mill/geographically indicated company. As only 2 samples were positive for aflatoxin, so contamination rate was 1.04%. In the meanwhile, contamination rate for ochratoxin A was 21.88% and a various amount of OTA was detected in 42 positive samples. 6 samples were found to be contaminated at higher level than $5\;{\mu}gkg^{-1}$ for ochratoxin A, which was established recently as a maximum permissible limit in korea. There was no difference in degree of contamination with regard to cultivation type because any mycotoxin was not found at all in both organically and conventionally grown red pepper. But, there was statistically significant difference in the process of manufacturing. Finished products were OTA-contaminated at a level of $2.32\;{\pm}\;6.54\;{\mu}gkg^{-1}$(mean ${\pm}$ SD), even though OTA was not detected in deep frozen red peppers right after long term storage. And contamination for OTA was a level of $0.33\;{\pm}\;0.91\;{\mu}gkg^{-1}$(mean ${\pm}$ SD) in red paprika powder after uv sterilization, while the contamination for OTA was $2.78\;{\pm}\;4.49\;{\mu}kg^{-1}$(mean ${\pm}$ SD) in non-uv sterilized powder. In addition, our investigation shows that higher OTA contamination occurred in some of famous brand products sold in super-market and domestic products than products collected through on-line shopping or from small stakeholder mills and imported products respectively, however, difference was not statistically significant.

• Journal of The Korean Society of Grassland and Forage Science
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• v.36 no.4
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• pp.333-339
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• 2016

The objective of this study was to optimize analytical methods for ochratoxin A (OTA) and zearalenone (ZEA) in rice straw silage and winter forage crops using ultra-high performance liquid chromatography (UHPLC). Samples free of mycotoxins were spiked with $50{\mu}g/kg$, $250{\mu}g/kg$, or $500{\mu}g/kg$ of OTA and $300{\mu}g/kg$, $1500{\mu}g/kg$, or $3000{\mu}g/kg$ of ZEA. OTA and ZEA were extracted by acetonitrile and cleaned-up using an immunoaffinity column. They were then subjected to analysis with UHPLC equipped with a fluorescence detector. The correlation coefficients of calibration curves showed high linearity ($R^2{\geq_-}0.9999$ for OTA and $R^2{\geq_-}0.9995$ for ZEA). The limit of detection and quantification were $0.1{\mu}g/kg$ and $0.3{\mu}g/kg$, respectively, for OTA and $5{\mu}g/kg$ and $16.7{\mu}g/kg$, respectively, for ZEA. The recovery and relative standard deviation (RSD) of OTA were as follows: rice straw = 84.23~95.33%, 2.59~4.77%; Italian ryegrass = 79.02~95%, 0.86~5.83%; barley = 74.93~97%, 0.85~9.19%; rye = 77.99~96.67%, 0.33~6.26%. The recovery and RSD of ZEA were: rice straw = 109.6~114.22%, 0.67~7.15%; Italian ryegrass = 98.01~109.44%, 1.65~4.81%; barley = 98~113.53%, 0.25~5.85%; rye = 90.44~108.56%, 2.5~4.66%. They both satisfied the standards of European Commission criteria (EC 401-2006) for quantitative analysis. These results showed that the optimized methods could be used for mycotoxin analysis of forages.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.82-87
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• 2007

Five dogs with renal failure were referred to the Veterinary Medical Teaching Hospital at Kangwon National University. These dogs had the common history of consumption of Pedigree dry dog food produced in Thailand plant for over 1 month. The dogs showed anorexia, emaciation, vomiting, and polydipsia/polyuria. And in one severely affected dog, bloody diarrhea and hypothermia were seen. The remarkable clinicopathological signs were high value of BUN and creatinine. In some dogs, GGT, phosphorus and lipase were increased. However, no significant changes of complete blood count were found. In urinalysis, hematuria, low specific gravity urine, proteinuria, and calcium oxalate-like crystals were observed. Two severely affected dogs were died. The remained dogs were recovered gradually after change of dog food and supportive therapy. Pathological findings were seen typically in kidneys. Renal atrophy, congestion of the glomerular capillary, and diffuse degeneration, necrosis, dystrophic calcification and regeneration in the tubular epithelium were seen. Yellowish brown fluorolucent laminated materials or particles were quite often found in the lumina of the necrotizing renal tubules of cortex and medulla. Proliferation of fibrous tissue in the interstitium was also seen. By the mycotoxin analysis of the Pedigree dry dog food, ochratoxin A (OTA) and citrinin were detected as much as the concentration of 372.8 ppb and 8.3 ppb, respectively. The final diagnosis of renal failure caused by OTA and citrinin toxicosis was made on the basis of history takings, clinical signs, clinicopathological and pathological findings, and analysis of mycotoxins.

• Korean Journal of Environmental Biology
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• v.31 no.3
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• pp.189-191
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• 2013

Mycotoxins such as aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) are widespread contaminants of food and feedstuffs. It is very likely, that humans and animals are always exposed to mixtures of mycotoxins rather than to individual compounds. Therefore, risk assessments should consider mixture toxicity data. In the present study the combination of AFB1, OTA and ZEA was tested for genotoxicity in rat bone marrow and blood leukocytes after 15, 30 and 60 days treatment. The level of DNA damage was determined by the comet assay. The tail intensity and Olive tail moment in leukocytes and bone marrow cells were significantly higher than in controls. At the same time, the level of DNA damage in bone marrow cells was higher than in leukocytes. The data suggests that prolonged exposure to mycotoxins combination through food consumption can induce DNA damage contributing to the harmful effects in vivo.