Vibrio gazogenes-dependent disruption of aflatoxin biosynthesis in Aspergillus flavus: the connection with endosomal uptake and hyphal morphogenesis.

Aflatoxins, a family of fungal secondary metabolites, are toxic and carcinogenic compounds that pose an enormous threat to global food safety and agricultural sustainability. Specifically agricultural products in African, Southeast Asian and hot and humid regions of American countries suffer most damage from aflatoxin producing molds due to the ideal climate conditions promoting their growth. Our recent studies suggest that Vibrio gazogenes (Vg), an estuarine bacterium non-pathogenic to plants and humans, can significantly inhibit aflatoxin biosynthesis in the producers. In this study, we investigated the mechanism underlying Vg-dependent aflatoxin inhibition using the prominent aflatoxin producer, Aspergillus flavus. We show that aflatoxin inhibition upon Vg treatment was associated with fungal uptake of Vg-prodigiosin, a red pigment, which was consistently visible inside fungal hyphae during treatment. The association of prodigiosin with aflatoxin inhibition was further evident as Serratia marcescens, another prodigiosin producer, significantly inhibited aflatoxin, while non-producers like Escherichia coli, Staphylococcus aureus, Vibrio harveyi, and Vibrio fischeri did not. Also, pure prodigiosin significantly inhibited aflatoxin biosynthesis. Endocytosis inhibitors, filipin and natamycin, reduced the Vg-prodigiosin uptake by the fungus leading to a significant increase in aflatoxin production, suggesting that uptake is endocytosis-dependent. The Vg treatment also reduced hyphal fusion (>98% inhibition) and branching, which are both endosome-dependent processes. Our results, therefore, collectively support our theory that Vg-associated aflatoxin inhibition is mediated by an endocytosis-dependent uptake of Vg-prodigiosin, which possibly leads to a disruption of normal endosomal functions.

Restricting mycotoxins without killing the producers: a new paradigm in nano-fungal interactions.

Over the past several years, numerous studies have demonstrated the feasibility of using engineered nanoparticles as antifungals, especially against those fungal pathogens that produce mycotoxins and infect plants, animals, and humans. The high dosage of nanoparticles has been a concern in such antifungal applications due to the potential toxicological and ecotoxicological impacts. To address such concerns, we have recently introduced the idea of inhibiting mycotoxin biosynthesis using low doses of engineered nanoparticles. At such low doses these particles are minimally toxic to humans and the environment. From our studies we realize that for the effective use of nanotechnology to intervene in the biology of fungal pathogens and for an accurate evaluation of the impacts of the increasingly growing nanomaterials in the environment on fungi and their interacting biotic partners, there is a pressing need for a rigorous understanding of nano-fungal interactions, which is currently far from complete. In this minireview, we build on the available evidence from nano-bio interaction research and our recent interaction studies with Aspergillus cells and engineered silver nanoparticles to introduce a potential theoretical model for nano-fungal interactions. The aim of the proposed model is to provide an initial insight on how nanoparticle uptake and their transformation inside fungal cells, possibly influence the production of mycotoxins and other secondary metabolites of filamentous fungi .

Acute and short-term administrations of delta-9-tetrahydrocannabinol modulate major gut metabolomic regulatory pathways in C57BL/6 mice.

Delta-9-tetrahydrocannabinol (THC) is the primary psychoactive compound in Cannabis, which is studied extensively for its medicinal value. A central gap in the science is the underlying mechanisms surrounding THC's therapeutic effects and the role of gut metabolite profiles. Using a mass-spectrometry based metabolomics, we show here that intraperitoneal injection of THC in C57BL/6 mice modulates metabolic profiles that have previously been identified as integral to health. Specifically, we investigated the effects of acute (single THC injection denoted here as '1X') and short -term (five THC injections on alternate days denoted as '5X') THC administration on fecal and intestinal tissue metabolite profiles. Results are consistent with the hypothesis that THC administration alters host metabolism by targeting two prominent lipid metabolism pathways: glycerophospholipid metabolism and fatty acid biosynthesis.

Size and coating of engineered silver nanoparticles determine their ability to growth-independently inhibit aflatoxin biosynthesis in Aspergillus parasiticus.

Recent studies from our laboratory indicate that engineered silver nanoparticles can inhibit aflatoxin biosynthesis even at concentrations at which they do not demonstrate antifungal activities on the aflatoxin-producing fungus. Whether such inhibition can be modified by altering the nanoparticles' physical properties remains unclear. In this study, we demonstrate that three differently sized citrated-coated silver nanoparticles denoted here as NP1, NP2, and NP3 (where, sizes of NP1 < NP2 < NP3) inhibit aflatoxin biosynthesis at different effective doses in Aspergillus parasiticus, the plant pathogenic filamentous fungus. Recapping NP2 with polyvinylpyrrolidone coating (denoted here as NP2p) also altered its ability to inhibit aflatoxin production. Dose-response experiments with NP concentrations ranging from 10 to 100 ng mL-1 indicated a non-monotonic relationship between aflatoxin inhibition and NP concentration. The maximum inhibitory concentrations differed between the NP types. NP1 demonstrated maximum inhibition at 25 ng mL-1. Both NP2 and NP3 showed maximum inhibition at 50 ng mL-1, although NP2 resulted in a significantly higher inhibition than NP3. While both NP2 and NP2p demonstrated greater aflatoxin inhibition than NP1 and NP3, NP2p inhibited aflatoxin over a significantly wider concentration range as compared to NP2. Our results, therefore, suggest that nano-fungal interactions can be regulated by altering certain NP physical properties. This concept can be used to design NPs for mycotoxin prevention optimally.

Complementary feeding may pose a risk of simultaneous exposures to aflatoxin M1 and deoxynivalenol in Indian infants and toddlers: Lessons from a mini-survey of food samples obtained from Kolkata, India.

A mini-survey of 29 different foods produced by 21 different Indian manufacturers was conducted for the presence of aflatoxins B1, B2, G1 and G2, aflatoxin M1 and deoxynivalenol. The products were purchased from local markets in Kolkata, India and commonly used in the complementary feeding of infants and toddlers in India. Using a previously established direct competitive enzyme-linked immunoassay for this analysis we show that 100% of the samples contained aflatoxin M1 at levels exceeding the recommended European Union levels of 25 ng kg-1 by more than an order of magnitude. Also, several (66%) of them contained detectable concentrations of deoxynivalenol with two samples (6.9%) exceeding European Union guidelines for baby food products (200 µg kg-1) and 51.7% samples with DON levels that can lead to dietary intake higher than 1  µg kg-1 recommended by the joint FAO/WHO expert committee on food additives. None of the samples contained aflatoxins B1, B2, G1 and G2. The results, therefore, suggest that complementary feeding can put Indian infants and toddlers at risk of simultaneous exposures to deoxynivalenol and aflatoxin M1 and warrant an urgent in-depth research to track, increase surveillance and reduce mycotoxin contamination of baby foods manufactured in India.