• Asian-Australasian Journal of Animal Sciences
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• 제16권5호
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• pp.706-713
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• 2003

Two experiments were conducted to evaluate the efficacy of dietary FERMKIT, a commercial toxin binder consisting of probiotic-fermented natural product containing chitin, chitosan and chitosan oligosaccharides ($FERMKITO^{(R)}$, EASY-BIO SYSTEM, Inc., Korea), in binding aflatoxin (AF) and zearalenone (ZEN) and ameliorating their mycotoxicity in meat type ducks. FERMKIT was supplemented to AF contaminated diets (at 120 ppb) at either 0.3 or 0.6% in experiment 1 and to ZEN contaminated diets (at 150 ppb) at 0.6% in experiment 2. In experiment 1 body weight gains were reduced by 37% and mortality was increased by 18% in ducks fed diet contaminated with AF at 120 ppb compared to ducks fed control diet (<10 ppb AF) for the 4-wk experimental period. However, dietary FERMKIT supplementation effectively alleviated overall toxicity induced by AF. The significant treatment-related changes in feather growth, web-toe hemorrhage, leg deformity, liver paleness, organ weights, hematological values and serum biochemical values, as compared to the control, were observed. The FERMKIT supplementation significantly diminished the adverse effects of AF and restored all the parameters measured back (<0.05) toward the control values. These findings indicated that FERMKIT, when added at the levels of 0.3 or 0.6% in the 120 ppb AF diets, could modulate the toxicity of AF with percentage sorption capacity of 52.70% at the level 0.3% and 79.85% at the level 0.6% of the diets (experiment 1). In experiment 2, FERMKIT, when added at 0.6% to the 150 ppb ZEN diets for the 4-wk experimental period, diminished the toxicity as shown by body weight gain, weights of testicles, oviducts, Bursa of Fabricius and cloaca eversion score as compared with the controls (<10 ppb ZEN) and 150 ppb ZEN diet with no added FERMKIT. The findings indicated that FERMKIT could be protective against the effects of ZEN in young growing ducks with percentage sorption capacity of 67.11% as evaluated from toxicity index parameter measured when added at 0.6% of the diets containing 150 ppb ZEN.

• Asian-Australasian Journal of Animal Sciences
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• 제18권9호
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• pp.1305-1309
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• 2005

Experiments were conducted to determine the efficacy of modified montmorillonite nanocomposite (MMN) to reduce the toxic effects of aflatoxin (AF) in growing/finishing pigs. 96 weaned pigs were assigned to four dietary treatment groups (0 g of MMN and 0 mg of AF/kg feed, 3 g of MMN/kg feed, 0.1 mg of AF/kg feed, and 3 g of MMN plus 0.1 mg of AF/kg feed). Body weight gain (BW gain), feed/gain ratio, serum biochemical values and enzyme activities were evaluated. Compared with the control, AF alone markedly reduced BW gain and resulted in a significantly higher feed/gain ratio. There were no differences in BW gain and feed/gain ratio between 0.3% MMN or 0.3% MMN plus AF and the control. These results suggested that the deleterious effects of AF were ameliorated by MMN addition. AF intake markedly increased relative organ weights of liver, kidney, spleen and pancreas, and resulted in significant alterations of serum parameters. However, these parameters for pigs fed diets containing MMN and AF returned to normal values, indicating that MMN had the ability to recover the AF-decreased performance, organ damage and to correct aberrations in serum parameters. These findings in our study suggested that MMN can effectively modulate the toxicity of AF in growing/finishing pigs and may offer a novel approach to the preventive management of aflatoxicosis in animals.

• Journal of Microbiology and Biotechnology
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• 제24권10호
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• pp.1397-1404
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• 2014

A few starters have been developed and used for doenjang fermentation but often without safety evaluation. Filamentous fungi were isolated from industrial doenjang koji, and their potential for mycotoxin production was evaluated. Two fungi were isolated; one was more dominantly present (90%). Both greenish (SNU-G) and whitish (SNU-W) fungi showed 97% and 95% internal transcribed spacer sequence identities to Aspergillus oryzae/flavus, respectively. However, the SmaI digestion pattern of their genomic DNA suggested that both belong to A. oryzae. Moreover, both fungi had morphological characteristics similar to that of A. oryzae. SNU-G and SNU-W did not form sclerotia, which is a typical characteristic of A. oryzae. Therefore, both fungi were identified to be A. oryzae. In aflatoxin gene cluster analysis, both fungi had norB-cypA genes similar to that of A. oryzae. Consistent with this, aflatoxins were not detected in SNU-G and SNU-W using ammonia vapor, TLC, and HPLC analyses. Both fungi seemed to have a whole cyclopiazonic acid (CPA) gene cluster based on PCR of the maoA, dmaT, and pks-nrps genes, which are key genes for CPA biosynthesis. However, CPA was not detected in TLC and HPLC analyses. Therefore, both fungi seem to be safe to use as doenjang koji starters and may be suitable fungal candidates for further development of starters for traditional doenjang fermentation.

• Toxicological Research
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• 제27권2호
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• pp.125-131
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• 2011

Through the present study, we produced a monoclonal antibody against aflatoxin B1 (AFB1) using AFB1-carboxymethoxylamine BSA conjugates. One clone showing high binding ability was selected and it was applied to develop a direct competitive ELISA system. The epitope densities of AFB1-CMO against BSA and KLH were about 1 : 6 and 1 : 545, respectively. The monoclonal antibody (mAb) from cloned hybridoma cell was the IgG1 subclass with ${\lambda}$-type light chains. The $IC_{50}s$ of the monoclonal antibody developed for AFB1, AFB2, AFG1 and AFG2 were 4.36, 7.22, 6.61 and 29.41 ng/ml, respectively, based on the AFB1-KLH coated ELISA system and 15.28, 26.62, 32.75 and 56.67 ng/ml, respectively, based on the mAb coated ELISA. Cross-relativities of mAb to AFB1 for AFB2, AFG1 and AFG2 were 60.47, 65.97 and 14.83% in the AFB1-KLH coated ELISA, and 59.41, 46.66 and 26.97% in the mAb coated ELISA, respectively. Quantitative calculations for AFB1 from the AFB1-Ab ELISA and AFB1-Ag ELISA ranged from 0.25 to 25 ng/ml ($R^2$ > 0.99) and from 1 to 100 ng/ml ($R^2$ > 0.99), respectively. The intra- and inter-assay precision CVs were < 10% in both ELISA assay, representing good reproducibility of developed assay. Recoveries ranged from 79.18 to 91.27%, CVs ranged from 3.21 to 7.97% after spiking AFB1 at concentrations ranging from 5 to 50 ng/ml and following by extraction with 70% methanol solution in the Ab-coated ELISA. In conclusion, we produced a group specific mAb against aflatoxins and developed two direct competitive ELISAs for the detection of AFB1 in feeds based on a monoclonal antibody developed.

• Journal of Preventive Medicine and Public Health
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• 제9권1호
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• pp.77-86
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• 1976

This study was attempted to know that the interactions of various fungi, and methionine and $MgSO_4$ introduced as the substrate of culture media for fungi were affected to produce aflatoxins by Asp. flavus. 5 different fungi were isolated from the fermented soybean mash and were cultured in Chemically Defined medium (C. D. media) and soybean mash at $25^{\circ}C$ for 10 days. (1) It was confirmed that Asp. flavus produced aflatoxins in the C. D. medium and soybean mast, but that Asp. niger, Asp. oryzae, Asp. awamori and Asp. terreus did not produced them respectively. (2) Asp. flavus cultured with Asp. niger did not produce aflatoxins in C. D. medium, but produced in soybean mash, in other hand, Asp. flavus with other fungi except Asp. niger produced aflatoxins in C. D. medium and soybean mash. (3) The growth of fungi were more prosperous in the seperate culture than in the mixed culture. (4) In the C. D. medium added 20% of cultured medium of Asp. niger, Asp. flavus did not produce aflatoxins but other cultured medium did not prohibit the production of aflatoxins by Asp. flavus. (5) On the contrary, $MgSO_4$ increasing the productivity of aflatoxins by Asp. flavus in the C. D. medium, methionine known as one of precursor of aflatoxins did not affected the increasing productivity with significance.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1740-1748
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• 2012

A home-made meju sample contaminated naturally with aflatoxins was used for isolation of fungal strains. Overall, 230 fungal isolates were obtained on dichloran rosebengal chloramphenicol (DRBC) and dichloran 18% glycerol (DG18) agar plates. Morphological characteristics and molecular analysis of a partial ${\beta}$-tubulin gene and the internal transcribed spacer (ITS) of rDNA were used for the identification of the isolates. The fungal isolates were divided into 7 genera: Aspergillus, Eurotium, Penicillium, Eupenicillium, Mucor, Lichtheimia, and Curvularia. Three strains from 56 isolates of the A. oryzae/flavus group were found to be aflatoxigenic A. flavus, by the presence of the aflatoxin biosynthesis genes and confirmatory aflatoxin production by high-performance liquid chromatography (HPLC). The predominant isolate from DRBC plates was A. oryzae (42 strains, 36.2%), whereas that from DG18 was A. candidus (61 strains, 53.5%). Out of the 230 isolates, the most common species was A. candidus (34.3%) followed by A. oryzae (22.2%), Mucor circinelloides (13.0%), P. polonicum (10.0%), A. tubingensis (4.8%), and L. ramosa (3.5%). A. flavus and E. chevalieri presented occurrence levels of 2.2%, respectively. The remaining isolates of A. unguis, P. oxalicum, Eupenicillium cinnamopurpureum, A. acidus, E. rubrum, P. chrysogenum, M. racemosus, and C. inaequalis had lower occurrence levels of < 2.0%.

• Asian-Australasian Journal of Animal Sciences
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• 제31권1호
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• pp.91-97
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• 2018

Objective: A 14-d trial was conducted to determine the effects of feeding corn naturally contaminated with aflatoxin $B_1$ ($AFB_1$) on growth performance, apparent ileal digestibility, serum hormones levels and gene expression of $Na^+$, $K^+-ATPase$ in ducklings. Methods: A total of 704 ducklings were blocked on the basis of sex and body weight (BW), and then allocated randomly to one of the following two treatments: i) CON, basal diet and ii) $AFB_1$, diets with 100% of normal corn replaced with $AFB_1$ contaminated corn. There were 22 pens per treatment and 16 birds per pen. The concentration of $AFB_1$ was 195.4 and $124.35{\mu}g/kg$ in the contaminated corn and $AFB_1$ diet, respectively. Results: The $AFB_1$ decreased average daily gain, average daily feed intake, d 7 BW, final BW in the whole trial, and feed conversion ratio (FCR) during d 8 to 14 and d 1 to 14 by 10% to 47% (p<0.05), while FCR during d 1 to 7 was increased (p<0.05). $AFB_1$ did not affect mortality to 7 d of age, and then increased to 5.8% from 8 to 14 d of age (p<0.01). Apparent ileal gross energy digestibility was reduced by $AFB_1$, whereas apparent ileal digestibility of dry matter, nitrogen, and amino acid was improved (p<0.01). Feeding $AFB_1$ diets increased serum concentration of leptin and insulin-like growth factors-1 (IGF-1) (p<0.05), but had no effect on neuropeptide Y, ghrelin, cholecystokinin-8 or insulin (p>0.05). Dietary treatments did not influence relative expression of jejunal $Na^+$, $K^+-ATPase$ gene (p>0.05). Conclusion: Taken together, feeding corn naturally contaminated with $AFB_1$ reduced growth performance, improved apparent ileal digestibility, and affected serum leptin and IGF-1 in ducklings from d 1 to 14.

• Interdisciplinary Bio Central
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• 제2권4호
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• pp.10.1-10.5
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• 2010

Total aflatoxin and ochratoxin were detected in 3 naturally contaminated fish feed samples using immune-affinity method. The results revealed that the average levels of aflatoxins in the 3 examined samples were (15, 22 and $12\;{\mu}g/kg$) respectively while the average levels of ochratoxins were (15, 6 and $6\;{\mu}g/kg$). The results of determination of the effects of clay as a mycotoxin binder on the health status and performance of Oreochromis niloticus in comparing with a control group revealed that the survival rate in control group was 81% after the end of the culture season. The results also revealed that the survival rate in group 2 which received clay treated feed was 86%. The results of regular parasitological examination revealed the identification of trichodina as external protozoa in Oreochromis niloticus from both ponds but without manifestation of disease signs. The results of bacteriological examination revealed the isolation and identification of Pseudomonas flouresence from some moribund Oreochromis niloticus. Higher performance parameters were recorded in group 2 that received feeds treated with clay which reflected in the total production which reaches 1646.47 kg while in the control pond, the total production was1308.36 kg.

• Mycobiology
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• 제35권2호
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• pp.47-53
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• 2007

The prevalence and population density of the mycobiota of 50 samples belonging to 10 kinds of spices (anise, black pepper, red pepper, black cumin, peppermint, cardamom, clove, cumin, ginger and marjoram) which collected from different places in Jeddah Governorate were studied. The natural occurrence of mycotoxins in those samples was also investigated. Fifteen genera and thirty-one species of fungi in addition to one species variety were isolated and identified during this study. The most common genera were Aspergillus, Penicillium and Fusarium. Aflatoxins ($12{\sim}40\;{\mu}g/kg$) were detected in the extract of 5 samples of each of anise seeds and black pepper fruits; three samples of black cumin seeds and on sample only of each of peppermint and marjoram leaves out of 5 samples tested of each. Sterigmatocystin ($15{\sim}20\;{\mu}g/kg$) was detected in some samples of red pepper, cumin and marjoram. The inhibitory effects of 10 kinds of powdered spices were tested against 3 toxigenic isolates of fungi (Aspergillus flavus, A. versicolor and Penicillium citrinum). Clove proved to be antimycotic compounds. It inhibited the growth of the tested toxigenic fungi. Black pepper, peppermint, cardamom, cumin and marjoram completely inhibited aflatoxins production, while black pepper and cardamom also completely inhibited sterigmatocystin production.

• 한국미생물·생명공학회지
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• 제47권2호
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• pp.234-241
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• 2019

In Korea, traditional doenjang is manufactured using the conventional method at home and by small-scale enterprises. Because this age-old process depends on natural inoculation of various microorganisms, it is difficult to reproduce or maintain consistency in the final product quality across batches. Moreover, doenjang occasionally prepared by this method raises safety concerns related to aflatoxin, biogenic amine, and Bacillus cereus contamination. To develop starters that can be conveniently used at home or in small industry settings for the manufacturing of safe and flavor-improved doenjang, autochthonous microbe starters were developed in dried forms as ready-to-use starters. Each starter powder prepared by heat- or freeze-drying methods remained stable even after 24-week storage. These ready- to-use starter powders were successfully applied to lab-scale fermentation for the production of safe and flavor-improved doenjang. We believe that these ready-to-use starter powders will benefit small-scale enterprises in the manufacturing of doenjang of good reproducible quality.