Calcium Montmorillonite for the Mitigation of Aflatoxicosis and Gastrointestinal Inflammation

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2014-08-06

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Abstract

Clays have been used for centuries as ?ancient medicine? for their therapeutic benefits. One particular clay, calcium montmorillonite, has historically been used as an anti-caking agent in animal feeds, but has also demonstrated the ability to bind toxins and alleviate infectious diarrhea. The full breadth of therapeutic applications and molecular mechanisms of montmorillonite is still unknown. Therefore, the purpose of this research was to explore novel therapeutic applications for NovaSil (NS), a calcium montmorillonite clay to reduce the risk of aflatoxicosis in farm-raised fish and alleviate gastrointestinal inflammation and dysbiosis in a mouse model of Crohn?s disease (CD).

Aflatoxin B_(1) (AFB_(1)) is a fungal mycotoxin that commonly contaminates corn and peanut crops. It is produced by the fungi Aspergillus flavus and A. parasiticus during times of drought or due to improper post-harvest storage. Aflatoxin B_(1) is known to cause hepatocellular carcinoma, immunosuppression and growth stunting in several species. Recently, incorporation of plant-based alternatives into feed for farm-raised fish has become a trend, thereby increasing the risk for mycotoxin contamination. Inexpensive strategies to reduce AFB_(1) exposure are needed. Calcium montmorillonite clay, which is both inexpensive and abundant, has a dioctahedral structure that is known to sequester AFB_(1) in its negatively-charged interlayer, thereby reducing systemic bioavailability. There is also some evidence to suggest that calcium montmorillonite clays may possess gastrointestinal anti-inflammatory properties.

NovaSil was used as a strategy to reduce the effects of AFB_(1) in Nile tilapia (Oreochromis niloticus) and red drum (Scieanops ocellatus). Juvenile tilapia and red drum were dosed with AFB_(1) and NS over the course of 10 and 7 weeks, respectively. Additionally, proinflammatory cytokine-clay binding was characterized using isothermal analysis, X-ray diffraction (XRD) and transmission electron microscopy (TEM). Furthermore, a TNBS (2,4,6-Trinitrobenzenesulfonic acid)-colitis gastrointestinal mouse model was employed to study the anti-inflammatory properties of NS and its ability to protect the gut microbiome.

Results suggest that NS can prevent aflatoxicosis in red drum at a 2% inclusion level over the course of 7 weeks. NovaSil also prevented some toxicity in Nile tilapia; however, these results were not significant. In vitro results also indicate that NS sorbs proinflammatory cytokines such as TNF? and IL-1? in its interlayers. Additionally, NS was found to reduce serum pro-inflammatory cytokine levels in TNBS-induced mice and reduce gut dysbiosis. These results could positively impact both human and animal populations with AFB_(1) exposure and/or chronic gastrointestinal inflammation.

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