3D Visualization of Developmental Toxicity of 2,4,6-Trinitrotoluene in Zebrafish Embryogenesis Using Light-Sheet Microscopy

Abstract

Environmental contamination by trinitrotoluene is of global concern due to its widespread use in military ordnance and commercial explosives. Despite known long-term persistence in groundwater and soil, the toxicological profile of trinitrotoluene and other explosive wastes have not been systematically measured using in vivo biological assays. Zebrafish embryos are ideal model vertebrates for high-throughput toxicity screening and live in vivo imaging due to their small size and transparency during embryogenesis. Here, we used Single Plane Illumination Microscopy (SPIM)/light sheet microscopy to assess the developmental toxicity of explosive-contaminated water in zebrafish embryos and report 2,4,6-trinitrotoluene-associated developmental abnormalities, including defects in heart formation and circulation, in 3D. Levels of apoptotic cell death were higher in the actively developing tissues of trinitrotoluene-treated embryos than controls. Live 3D imaging of heart tube development at cellular resolution by light-sheet microscopy revealed trinitrotoluene-associated cardiac toxicity, including hypoplastic heart chamber formation and cardiac looping defects, while the real time PCR (polymerase chain reaction) quantitatively measured the molecular changes in the heart and blood development supporting the developmental defects at the molecular level. Identification of cellular toxicity in zebrafish using the state-of-the-art 3D imaging system could form the basis of a sensitive biosensor for environmental contaminants and be further valued by combining it with molecular analysis.

Developmental toxicity of dimethachlor during zebrafish embryogenesis mediated by apoptosis and oxidative stress

“…This result is in conjunction with body length shortening by dimethachlor. Also, elevated apoptosis might result in defects in the vascular and neural systems in the posterior regions, which negatively affects whole embryogenesis (Eum et al, 2016;Chen et al, 2017). Dimethachlor generated not only apoptotic cells but also ROS, which adversely affected embryonic organogenesis.…”

Biodegradation of 2,4,6-Trinitrotoluene (TNT) with Bacteria Isolated from TNT-polluted Waste Pink Water

“…TNT and its associated explosive compounds cause pollution if this water is delivered to the soil without purification [5]. TNT contamination from TNT-containing pink water in military regions has also been reported by researchers in the literature [5,7]. Chusova et al [5] collected TNT contaminated pink water released from a treatment plant at the demilitarisation factory Nammo Vingakersverken AB and they stated that the TNT level in the pink water was 12mM after serial dilutions (5 times) and removing of the insoluble components.…”

“…Chusova et al [5] collected TNT contaminated pink water released from a treatment plant at the demilitarisation factory Nammo Vingakersverken AB and they stated that the TNT level in the pink water was 12mM after serial dilutions (5 times) and removing of the insoluble components. Eum et al [7] collected pink water from Agency for Defense Development, Daejeon, Korea for researching the effects of TNT-contaminated pink water in Zebrafish embryogenesis. They stated that the amount of TNT in the pink water was 45.54 mg/L.…”

Intensification of the leachate treatment process of nitrocellulose production

“…Nitrocellulose industry wastewater is highly toxic to the environment because it contains chlorophenols, chlorolignins, organic acids, acid resins, and dioxins [23,32], as well as high levels of organic matter and metal ions [22]. There are few studies analyzing the toxicity of nitrocellulose to aquatic organisms [37-39], but many studies reported the detrimental effects of other explosives, such as 2,4,6-trinitrotoluene (TNT), 1,3,5-Trinitroperhydro-1,3,5-triazine (RDX), and 1,3,5,7tetranitro-1,3,5,7-tetrazocane (HMX) [40][41][42][43][44][45][46].…”