Naiyi Yin

In Vitro Method To Assess Soil Arsenic Metabolism by Human Gut Microbiota: Arsenic Speciation and Distribution

Arsenic (As) speciation and distribution are two important factors in assessing human health risk from As-contaminated soil. In this study, we used the combination of physiologically based extraction test (PBET) and Simulator of Human Intestinal Microbial Ecosystem (SHIME) to determine soil As metabolism by human gut microbiota. The results showed that the percentage of soil arsenate [As(V)] transformation reached 22.1-38.2%, while that of arsenite [As(III)] attained 66.5-92.0%; 30.1-56.4% of As(V) transformed was attached to the soil solid phase. In comparison to sequential extraction results, almost all amorphous Fe/Al-oxide-bound As was liberated in the colon phase. An X-ray absorption near-edge structure (XANES) showed that the As(III) percentage in the soil solid phase reached 16.6-26.9% and reached 73.4% (soil 1) in the colon phase. Additionally, plenty of As(III) and different extents of methylation were also observed in colon extraction solution. As bioaccessibility in the colon phase was 1.8-2.8 times that in the small intestinal phase. Our results indicated that human gut microbiota increased As bioaccessibility, and large amounts of As(III) were adsorbed onto the soil solid phase as a result of microbial reduction. Determining As speciation and distribution in extraction solution and soil solid phases will allow for an accurate assessment of the risk to human health upon soil As exposure.

Variability of arsenic bioaccessibility and metabolism in soils by human gut microbiota using different in vitro methods combined with SHIME.

Arsenic (As) speciation analysis is essential when evaluating the risks upon oral exposure to As-contaminated soils. In this study, we first investigated the variability in the As bioaccessibility and speciation using a combination of five common in vitro methods (SBRC, PBET, DIN, UBM and IVG) (gastric and small intestinal phases) and the SHIME model (colon phase). Our results indicate that the As bioaccessibility varies in the colon phase. An increase in the As bioaccessibility for SBRC and PBET, and a decrease for UBM and IVG were observed in the colon phase. In addition, we found different extents of methylation and large amounts of arsenite [As(III)] due to microbial reduction in the colon digests. The UBM-SHIME method displayed a higher methylation percentage of 13.5-82.1%, but a lower methylation percentage of 0.2-21.8% was observed in the SBRC-SHIME method. Besides, The MMA(V) levels in the colon digests were positively correlated with those of As(III) and DMA(V), so DMA(V) can be considered an indicator to evaluate the As metabolic speed of in vitro cultured human gut microbiota. Based on the standard reference soil of NIST 2710a, the As bioaccessibility in the colon phase of PBET-SHIME and SBRC-SHIME were the closest to the in vivo results. Combining in vitro methods and SHIME will remarkably affect the accurate assessment of potential risks to human health associated with oral exposure to soil As.

Arsenic redox transformation by Pseudomonas sp. HN-2 isolated from arsenic-contaminated soil in Hunan, China

A mesophilic, Gram-negative, arsenite[As(III)]-oxidizing and arsenate[As(V)]-reducing bacterial strain, Pseudomonas sp. HN-2, was isolated from an As-contaminated soil. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that the strain was closely related to Pseudomonas stutzeri. Under aerobic conditions, this strain oxidized 92.0% (61.4μmol/L) of arsenite to arsenate within 3hr of incubation. Reduction of As(V) to As(III) occurred in anoxic conditions. Pseudomonas sp. HN-2 is among the first soil bacteria shown to be capable of both aerobic As(III) oxidation and anoxic As(V) reduction. The strain, as an efficient As(III) oxidizer and As(V) reducer in Pseudomonas, has the potential to impact arsenic mobility in both anoxic and aerobic environments, and has potential application in As remediation processes.

Formation of silver nanoparticles by human gut microbiota

Metal nanoparticles have received global attention and one of the most promising nanotechnologies is the use of microorganisms for synthesizing nanoparticles. The oral exposure study has demonstrated the formation of silver (Ag) nanoparticles with in vivo animal models. However, there is limitation in the effect of in vitro cultured human gut microbiota on silver. Here, the phenomenon of biosynthesis of Ag nanoparticles by human gut microbiota has been reported. The size, shape, and morphology of the nanoparticles were characterized using transmission electron microscopy (TEM). Exposure to aqueous AgNO3 solution resulted in the intracellular reduction of Ag+ ions and the formation of approximately spherical Ag nanoparticles with dimensions of 34 ± 10 nm. Electron microscopy analysis of the gut microbial cells by TEM images of thin sections and scanning electron microscopy (SEM), indicated that most of Ag nanoparticles were located on the cytoplasmic membrane and within the cytoplasm, where abundant silver reductases were present. In addition, some nanoparticles were bound to the cell wall, likely via exocytosis. The oral exposure to Ag+ led to in vivo formation of nanoparticles by gut microbiota, and their biological effects and potential applications in human body warrant further investigation.

Nutritional status affects the bioaccessibility and speciation of arsenic from soils in a simulator of the human intestinal microbial ecosystem

Arsenic (As) is a highly toxic contaminant in food and soil. In this study, we investigated the effects of four nutritional states (including a fed state with vitamin C, a fed state with protein powder, a fed state with glucose and a fasted state) on the variability of soil As bioaccessibility and biotransformation using the physiologically based extraction test (PBET) combined with a simulator of the human intestinal microbial ecosystem model (SHIME). The results indicated that the vitamin C and protein powder increased As bioaccessibility in gastric digests. In the colon phase, As bioaccessibility was observably enhanced by protein powder, and it varied under the vitamin C and glucose conditions. Additionally, the order of As methylation percentages in the four nutritional states was protein powder > vitamin C > fasted state > glucose (except S2); As bioaccessibility increased 1.3-13.7% and 15.8-35.4% in treatments of the vitamin C and protein powder, respectively. Meanwhile, large amounts of monomethylarsonic acid (MMAV) were observed in the colon digest in the protein powder condition. In contrast, As methylation was significantly decreased with the addition of glucose, with a decline of 25.9-45.5%. Additionally, glucose enhanced the reduction of As(V). Therefore, nutritional status is a crucial parameter for the prediction of bioaccessibility and speciation of As when assessing health risks from As following oral exposure.

Variability of Chromium Bioaccessibility and Speciation in Vegetables: The Influence of In Vitro Methods, Gut Microbiota and Vegetable Species

There is limited research concentrating on the effects of gut microbiota on the bioaccessibility and speciation of chromium (Cr) in vegetables. In this study, the physiologically based extraction test (PBET) and the unified BARGE method (UBM), were combined with the simulator of human intestinal microbial ecosystems (SHIME) to determine the bioaccessibility and speciation of Cr from vegetables. The results showed that the Cr bioaccessibility was the highest in the gastric phase. The Cr bioaccessibility from the water spinach was the highest, and was 1.6-3.4 and 1.1-1.8 times that of leaf lettuce and celery, respectively. The Cr bioaccessibilities of the UBM method were slightly greater than those of the PBET method. Additionally, the gut microbiota increased the Cr bioaccessibility and reduced more toxic Cr(VI) to less toxic Cr(III) from vegetables. Therefore, our study reveals the possible health risks of consuming Cr-contaminated vegetables based on the bioaccessibility and speciation of Cr.

Investigation of bioaccessibility of Cu, Fe, Mn, and Zn in market vegetables in the colon using PBET combined with SHIME

The in vitro bioaccessibility of trace metals associated with oral ingestion of market vegetables (lettuce, pak choi, cole, and leaf lettuce) of Beijing, China was studied. The physiologically based extraction test (PBET) combined with the Simulator of Human Intestinal Microbial Ecosystem (SHIME) was applied to simulate stomach, small intestine, and colon of human. In the gastro-intestinal phases, the bioaccessibility of Cu, Fe, Mn, and Zn varied within 5.7–75.5%, 17.3–50.4%, 13.3–49.1%, and 19.9–63.7%, respectively. There was no significant difference in the metal bioaccessibility between the gastric and small intestinal phases, except for higher Cu bioaccessibility in the small intestine. Besides, the bioaccessibility of the four trace metals in the colon phase was first ever reported. A significant decline in Cu bioaccessibility (1.8–63.7%) and slight increases in the bioaccessibility of Fe (16.7–56.4%), Mn (21.2–71.6%), and Zn (15.7–69.7%) were revealed, which could mainly be attributed to the effect of colon microbiota. In addition, the estimated daily intakes (EDIs) of Cu, Fe, Mn, and Zn were worked out to be 0.7, 8.8, 2.7, and 4.5 μg kg−1 body weight d−1, based on which the potential influences of these trace metals in vegetables on the health of the local consumers was demonstrated.