Toxicity of mycotoxins in vivo on vertebrate organisms: A review.

The EFSA Panel on Contaminants in the Food Chain (CONTAM) established a tolerable daily intake (TDI) for fumonisin B1 (FB 1) of 1.0 μg/kg body weight (bw) per day based on increased incidence of megalocytic hepatocytes found in a chronic study with mice. The CONTAM Panel considered the limited data available on toxicity and mode of action and structural similarities of FB 2-6 and found it appropriate to include FB 2, FB 3 and FB 4 in a group TDI with FB 1. Modified forms of FBs are phase I and phase II metabolites formed in fungi, infested plants or farm animals. Modified forms also arise from food or feed processing, and include covalent adducts with matrix constituents. Non-covalently bound forms are not considered as modified forms. Modified forms of FBs identified are hydrolysed FB 1-4 (HFB 1-4), partially hydrolysed FB 1-2 (pHFB 1-2), N-(carboxymethyl)-FB 1-3 (NCM-FB 1-3), N-(1-deoxy-d-fructos-1-yl)-FB 1 (NDF-FB 1), O-fatty acyl FB 1, N-fatty acyl FB 1 and N-palmitoyl-HFB 1. HFB 1, pHFB 1, NCM-FB 1 and NDF-FB 1 show a similar toxicological profile but are less potent than FB 1. Although in vitro data shows that N-fatty acyl FBs are more toxic in vitro than FB 1, no in vivo data were available for N-fatty acyl FBs and O-fatty acyl FBs. The CONTAM Panel concluded that it was not appropriate to include modified FBs in the group TDI for FB 1-4. The uncertainty associated with the present assessment is high, but could be reduced provided more data are made available on occurrence, toxicokinetics and toxicity of FB 2-6 and modified forms of FB 1-4.

https://efsa.onlinelibrary.wiley.com/doi/pdf/10.2903/j.efsa.2018.5172

Appropriateness to set a group health‐based guidance value for fumonisins and their modified forms

Chemistry, bioactivities, extraction and analysis of azadirachtin: State-of-the-art.

Factors affecting content and bioefficacy of neem (Azadirachta indica A. Juss.) phytochemicals used in agricultural pest control: A review

Synthesis and characterization of newly cationic surfactants based on 2-(2-(dimethylamino)ethoxy)ethanol: physiochemical, thermodynamic and evaluation as biocide

The effect of surfactants (two cationic, one anionic and three non-ionic) at 0.001, 0.01, 0.1 and 1.0 % concentrations on aflatoxin production, ergosterol content and sugar consumption by Aspergillus parasiticus (NRRL 2999) in YES liquid culture medium is reported. At 0.01% concentration, the cationic surfactants, cetyl dimethyl ammonium bromide (CDAB) and dodecyl trimethyl ammonium bromide (DTAB), and the anionic surfactant, sodium dodecyl sulfate (SDS), completely inhibited spore germination, while DTAB also inhibited the production of ergosterol and toxin (p < 0.05). At a concentration of 0.001%, CDAB was found to enhance toxin production, while SDS was found to inhibit it when compared with other surfactants. Non-ionic surfactants, polyoxyethylene sorbitan monolaurate (Tween-20), polyoxyethylene lauryl ether (Brij-35) and ethoxylated p-tert-octylphenol (Triton X-100) delayed the spore germination up to day 5 at all concentrations and inhibited toxin and ergosterol production at 0.001% concentration. The affect was found to be dose-dependent from 0.001% to 1%, for Triton X-100 only. Positive correlation between ergosterol content and toxin production in the presence of different surfactants at various time periods (3, 5, 7, 9 and 12 days) was found. Tween-20 was most effective in inhibiting toxin production on day 7, when aflatoxin production was found to be maximal in control group. Sugar consumption was directly proportional to the ergosterol content, showing a significant correlation with aflatoxin production.

Effect of various surfactants (cationic, anionic and non-ionic) on the growth of Aspergillus parasiticus (NRRL 2999) in relation to aflatoxin production

The genotoxic potential of fumonisin B1 (FB1) in vivo in BALB/c mice (male and female) was assessed by induction of micronuclei (MN) formation in bone marrow polychromatic erythrocytes. The ratio of polychromatic erythrocytes to normochromatic erythrocytes (PCE/NCE) was also determined. Mice were injected intraperitoneally (i.p.) with FB1 at a low dose (0.1 mg kg−1 body mass), middle dose (1.0 mg kg−1 body mass) and high dose (10 mg kg−1 body mass) as single and multiple doses in normal saline to test the genotoxicity. Mitomycin C, a known clastogen, was used as positive control. The frequency of MN and the PCE/NCE value in animals treated with FB1 at low, middle, and high doses in single dose studies, and the frequency of MN in multiple dose studies, were statistically non-significant from that of the controls injected with saline only. The multiple dose studies at all doses revealed that the PCE/NCE value was found to be reduced upon exposure to FB1 as compared to the controls. In animals injected with multiple low doses of FB1, the PCE/NCE value was found to be 0.66 in males and 0.82 in the females; at multiple middle doses the value was 0.30 in males and 0.41 in the females and was statistically significant (p < 0.001); however, at multiple high doses, the ratio was found to be 0.36 in both males and females. The present study confirms that FB1 is non-genotoxic in nature while the reduced ratio of PCE/NCE suggests the cytotoxic nature of FB1.

Lack of micronuclei induction by fumonisin B(1) mycotoxin in BALB/c mice.

Neem seed kernels artificially infested with Aspergillus parasiticus (NRRL 2999) was evaluated for aflatoxin elaboration and fungal growth, and compared with groundnut, a high risk commodity for aflatoxin contamination. At optimal moisture content (10-12%) the total, individual toxins (AFB1, AFB2, AFG1 and AFG2) and ergosterol content increased and showed maximum levels on day 9. Crude protein and polyphenols increased while, fat and total sugar content decreased during the period of infection. The protein content correlated positively (r = 0.734) with total toxin levels, whereas fat content (r = -0.761) and total sugars (r = -0.891) showed negative correlation and they were all statistically significant (p<0.01). The polyphenols showed negative and non-significant correlation with total toxin levels. Azadirachtin one of the major active principles of neem seed kernel showed significant decrease on day 3 (P<0.05) and day 6 (P<0.01). Neem seed kernel has shown 54 and 74% less aflatoxin production on day 9 and 12, respectively in comparison to groundnut seeds. Ergosterol content also showed 60% decrease on day 9, conferring it a poor substrate for fungal growth and aflatoxin elaboration.

http://nopr.niscair.res.in/bitstream/123456789/14821/1/IJNPR%203%283%29%20395-406.pdf

Substrate suitability of neem seed kernel for the growth and elaboration of aflatoxins by Aspergillus parasiticus (NRRL 2999)

The effect of neem seed kernel extracts and purified terpenoids on Aspergillus parasiticus in relation to fungal growth, ergosterol synthesis and aflatoxin elaboration was investigated in yeast extract sucrose (YES) liquid medium. Aqueous (0.05, 0.25, 0.5 and 1.0 %), hexane, methanolic, ethyl acetate and acid-methanolic extracts (0.01, 0.1, 0.5 and 1.0 %) of neem seed kernel were tested at 3, 5, 7, 9 and 12 days of incubation. Fungal growth was not affected by aqueous and hexane extracts, while it was inhibited by the methanolic, ethyl acetate and acid-methanolic extracts. The inhibition of ergosterol biosynthesis was 73 and 82 % by the aqueous extract at 0.5 and 1.0 % conc, while hexane and ethyl acetate inhibited in a dose dependent manner on day 7. The AFB 1 /total toxin altered significantly ( p <0.01) in the presence of various neem seed kernel extracts, except that of hexane extract. Purified terpenoids, azadirachtin, nimbin and salannin had no inhibitory effect on fungal biomass and aflatoxin elaboration at 0.01, 0.1, 0.25 and 0.5 % levels. Azadirachtin at 0.25 and 0.5 % level inhibited ergosterol biosynthesis by 30 and 34 %, respectively. Neem seed kernel extracts were inhibitive to the fungal growth, ergosterol biosynthesis and toxin elaboration as compared to purified neem terpenoids, suggesting anti-fungal and anti-aflatoxigenic properties.

http://nopr.niscair.res.in/bitstream/123456789/27472/1/IJNPR%205%281%29%2020-33.pdf

Inhibitory role of neem seed kernel extracts and terpenoids on growth and aflatoxin production by Aspergillus parasiticus (NRRL 2999)

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Effect of various surfactants (cationic, anionic and non-ionic) on the growth of Aspergillus parasiticus (NRRL 2999) in relation to aflatoxin production

Lack of micronuclei induction by fumonisin B(1) mycotoxin in BALB/c mice.

Substrate suitability of neem seed kernel for the growth and elaboration of aflatoxins by Aspergillus parasiticus (NRRL 2999)

Inhibitory role of neem seed kernel extracts and terpenoids on growth and aflatoxin production by Aspergillus parasiticus (NRRL 2999)