Huahong Shi

Microplastic Pollution in Table Salts from China

Microplastics have been found in seas all over the world. We hypothesize that sea salts might contain microplastics, because they are directly supplied by seawater. To test our hypothesis, we collected 15 brands of sea salts, lake salts, and rock/well salts from supermarkets throughout China. The microplastics content was 550-681 particles/kg in sea salts, 43-364 particles/kg in lake salts, and 7-204 particles/kg in rock/well salts. In sea salts, fragments and fibers were the prevalent types of particles compared with pellets and sheets. Microplastics measuring less than 200 μm represented the majority of the particles, accounting for 55% of the total microplastics, and the most common microplastics were polyethylene terephthalate, followed by polyethylene and cellophane in sea salts. The abundance of microplastics in sea salts was significantly higher than that in lake salts and rock/well salts. This result indicates that sea products, such as sea salts, are contaminated by microplastics. To the best of our knowledge, this is the first report on microplastic pollution in abiotic sea products.

Microplastics and mesoplastics in fish from coastal and fresh waters of China

Plastic pollution is a growing global concern. In the present study, we investigated plastic pollution in 21 species of sea fish and 6 species of freshwater fish from China. All of the species were found to ingest micro- or mesoplastics. The average abundance of microplastics varied from 1.1 to 7.2 items by individual and 0.2-17.2 items by gram. The average abundance of mesoplastics varied from 0.2 to 3.0 items by individual and 0.1-3.9 items by gram. Microplastics were abundant in 26 species, accounting for 55.9-92.3% of the total number of plastics items in each species. Thamnaconus septentrionalis contained the highest abundance of microplastics (7.2 items/individual). The average abundance of plastics in sea benthopelagic fishes was significantly higher than in freshwater benthopelagic fishes by items/individual. The plastics were dominanted by fiber in shape, transparent in color and cellophane in composition. The proportion of plastics in the stomach to the intestines showed great variation in different species, ranging from 0.5 to 1.9 by items/individual. The stomach of Harpodon nehereus and intestines of Pampus cinereus contained the highest number of plastics, (3.3) and (2.7), respectively, by items/individual. Our results suggested that plastic pollution was widespread in the investigated fish species and showed higher abundance in comparison with worldwide studies. The ingestion of plastics in fish was closely related to the habitat and gastrointestinal tract structure. We highly recommend that the entire gastrointestinal tract and digestion process be used in future investigations of plastic pollution in fish.

Electron paramagnetic resonance investigation of in vivo free radical formation and oxidative stress induced by 2,4-dichlorophenol in the freshwater fish Carassius auratus

In the present study, electron paramagnetic resonance coupled with spin-trapping technique was used, with alpha-phenyl-N-tert-butylnitrone (PBN) as a spin-trapping agent, to investigate free radical generation in freshwater fish with acute 2,4-dichlorophenol (2,4-DCP) poisoning. The PBN-radical adducts were detected in fish liver samples following treatments of 2,4-DCP (0.025, 0.05, 0.5, 5, or 25 mg/kg) 24 h after intraperitoneal (i.p.) injection and 2,4-DCP (0.5 mg/kg) at 2, 4, 8, 24, or 72 h after i.p. injection in Carassius auratus. The hyperfine splitting constants for the PBN-radical adducts are aN = 13.7 G, aH = 1.8 G, and g = 2.0058, which is consistent with those of PBN/hydroxyl radical (*OH). The results indicate that the hydroxyl radical is probably produced during acute intoxication of 2,4-DCP. The relative similarity in the kinetics (from 2 to 72 h) of superoxide dismutase activity induction and *OH generation implies that the generation of *OH possibly depends on the superoxide anion (O2*-). Superoxide anion (O2*-) might be the precursor radical undergoing the Haber-Weiss reaction to form *OH. Possible pathways for radical chain reactions in the formation of the hydroxyl radical in vivo after 2,4-DCP administration are proposed. Other parameters with respect to antioxidant defense (e.g., superoxide dismutase and catalase) and oxidative damage (lipid peroxidation level) indicate that the fish were subjected to oxidative stress induced by 2,4-DCP and that the mechanisms of oxidative stress possibly involve the in vivo stimulation of hydroxyl radical formation.

Effects of tributyltin (TBT) on Xenopus tropicalis embryos at environmentally relevant concentrations

Tributyltin (TBT) has been widely used as a biocide in antifouling paints and is a known endocrine disrupting chemical. In this paper, we exposed embryos of Xenopus tropicalis to 50-400ngL(-1) tributyltin chloride. TBT significantly decreased the survival rate, reduced the body length and retarded the development of embryos after 24, 36 and 48h of exposure. These effects of TBT were concentration- and time-dependent. Embryos treated with TBT showed multiple malformations. The most obvious alterations were abnormal eyes, enlarged proctodaeum, narrow fins, and skin hypopigmentation. Enlarged proctodaeum and narrow fins were mainly observed after 36 and 48h of exposure. The loss of eye pigmentation or the absence of external eyes occurred after 24 and 36h of exposure, while extended lenses or edemas of eyes were more commonly observed after 48h of exposure. Additional malformations included: small anterior region of heads, pericardial edemas, enlarged trunks, and bent tails. These results suggested that TBT is very toxic to X. tropicalis embryos at environmentally relevant concentrations.

Teratogenic effects of triphenyltin on embryos of amphibian (Xenopus tropicalis): A phenotypic comparison with the retinoid X and retinoic acid receptor ligands

Triphenyltin (TPT) has high binding affinity with the retinoid X receptor (RXR) in animals. The natural ligand of RXR, 9-cis-retinoic acid (RA), is known to induce featured malformations in vertebrate embryos by disrupting RA signal. Limited information is available on the TPT effects on amphibians. We exposed embryos of amphibian (Xenopus tropicalis) to TPT, 9-cis-RA, all-trans-RA (ligand of retinoic acid receptor, RAR), and LGD1069 (a selective ligand of RXR). The 72h LC50 of TPT was 5.25 μg Sn/L, and 72h EC50 was 0.96 μg Sn/L. TPT induced multiple malformations including enlarged proctodaeum and narrow fins. TPT at 5 μg Sn/L inhibited the differentiation of skins and muscles. The reduced brain, loss of external eyes and bent axis were observed in RXR and RAR ligands treatments. TPT and tributyltin (TBT) inhibited the mRNA expression of RXRα and increased that of TRβ. The phenotypes of malformations induced by TPT were similar to those by TBT and were much different from those by the RXR and RAR ligands. These results indicated that TPT was acute toxic and had high teratogenicity to amphibian embryos, and that TPT induced phenotypes of malformations. TPT and TBT might have a similar teratogenic mechanism, which seems not to be mainly mediated through RA signal.

Microplastic particles cause intestinal damage and other adverse effects in zebrafish Danio rerio and nematode Caenorhabditis elegans

Microplastics have been frequently detected in aquatic environments, and there are increasing concerns about potential effects on biota. In this study, zebrafish Danio rerio and nematode Caenorhabditis elegans were used as model organisms for microplastic exposure in freshwater pelagic (i.e. water column) and benthic (i.e. sediment) environments. We investigated the toxic effects of five common types of microplastics: polyamides (PA), polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC) and polystyrene (PS) particles. Results showed no or low lethality in D. rerio after exposure for 10d at 0.001-10.0mgL-1 microplastics. The PA, PE, PP and/or PVC microplastics with ~70μm size caused intestinal damage including cracking of villi and splitting of enterocytes. Exposure to 5.0mgm-2 microplastics for 2d significantly inhibited survival rates, body length and reproduction of C. elegans. Moreover, exposure to microplastics reduced calcium levels but increased expression of the glutathione S-transferase 4 enzyme in the intestine, which indicates intestinal damage and oxidative stress are major effects of microplastic exposure. Among 0.1, 1.0 and 5.0μm sizes of fluorescently labeled PS, 1.0μm particles caused the highest lethality, the maximum accumulation, the lowest Ca2+ level in the intestine and the highest expression of glutathione S-transferase 4 in nematodes. Taken together, these findings suggest that intestinal damage is a key effect of microplastics; and that the toxicity of microplastics is closely dependent on their size, rather than their composition.

Teratogenic effects of tetrabromobisphenol A on Xenopus tropicalis embryos.

Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardant and a known thyroid disruptor. We reported exposing Xenopus tropicalis embryos (NF10) to 0.01, 0.1 or 1 mg/L of TBBPA with or without 70 microg/L triiodothyronine (T(3)). Compared with the controls, 1 mg/L of TBBPA significantly reduced the body length of embryos after 24, 36 and 48 h of exposure. Embryos treated with TBBPA showed multiple malformations, including: abnormal eyes, skin hypopigmentation, enlarged proctodaeum, narrow fins and pericardial edemas. The effect of abnormal eyes manifested itself in the loss of pigmentation, reduction in size, or absence of external eyes. The degree of eye malformations was quantified with the index of eye malformations (IEM) with 0 being normal and 3 being severe. In the 1 mg/L TBBPA treatment groups, the incidence of total malformations (ITM) was 68-93%, and IEM was 0.8-0.9. T(3) showed no teratogenic effects on embryos, but it significantly enhanced TBBPA-induced teratogenic effects. In the T(3)+1 mg/L TBBPA treatment groups, ITM was 91-99%, and IEM was 1.8-1.9. Histological observations showed that the retinas were generally smaller, and the lenses were underdeveloped or even absent. These results indicate that TBBPA at relatively high concentration has teratogenic effects on X. tropicalis embryos. The results also suggest that thyroid hormone signaling might be involved in TBBPA induced-teratogenicity.

Adherence of microplastics to soft tissue of mussels: A novel way to uptake microplastics beyond ingestion

Microplastic pollution is recognized as an emerging threat to aquatic ecosystems. One of the main environmental risks associated with microplastics is their bioavailability to marine organisms. Up to date, ingestion has been widely accepted as the sole way for the animals to uptake microplastics. Nevertheless, microplastics have also been found in some organs which are not involved in the process of ingestion. We hypothesize that the animal might uptake microplastics through adherence in addition to ingestion. To test this hypothesis, we collected mussels from the fishery farms, conducted exposure/clearance experiments and analyzed the accumulation of microplastics in specific organ of mussels. Our studies clearly showed the uptake of microplastic in multiple organs of mussels. In the field investigations, we found that the abundance of microplastic by weight but not by individual showed significant difference among organs, and the intestine contained the highest level of microplastics (9.2items/g). In the uptake and clearance experiment, the accumulation and retention of microfibers could also be observed in all tested organs of mussels including foot and mantle. Our results strongly suggest that adherence rather than ingestion led to the accumulation of microplastics in those organs which are not involved in ingestion process. To our best knowledge, it is the first time to propose that adherence is a novel way for animals to uptake microplastics beyond ingestion. This new finding makes us rethink about the bioavailability, accumulation and toxicity of microplastics to aquatic animals.

Strong lethality and teratogenicity of strobilurins on Xenopus tropicalis embryos: Basing on ten agricultural fungicides

Agricultural chemical inputs have been considered as a risk factor for the global declines in amphibian populations, yet the application of agricultural fungicides has increased dramatically in recent years. Currently little is known about the potential toxicity of fungicides on the embryos of amphibians. We studied the effects of ten commonly used fungicides (four strobilurins, two SDHIs, two triazoles, fludioxonil and folpet) on Xenopus tropicalis embryos. Lethal and teratogenic effects were respectively examined after 48 h exposure. The median lethal concentrations (LC50s) and the median teratogenic concentrations (TC50s) were determined in line with actual exposure concentrations. These fungicides except two triazoles showed obvious lethal effects on embryos; however LC50s of four strobilurins were the lowest and in the range of 6.81-196.59 μg/L. Strobilurins, SDHIs and fludioxonil induced severe malformations in embryos. Among the ten fungicides, the lowest TC50s were observed for four strobilurins in the range of 0.61-84.13 μg/L. The teratogenicity shared similar dose-effect relationship and consistent phenotypes mainly including microcephaly, hypopigmentation, somite segmentation and narrow fins. The findings indicate that the developmental toxicity of currently-used fungicides involved with ecologic risks on amphibians. Especially strobilurins are highly toxic to amphibian embryos at μg/L level, which is close to environmentally relevant concentrations.

Effects of tributyltin on metamorphosis and gonadal differentiation of Xenopus laevis at environmentally relevant concentrations

Tributyltin (TBT), a well known endocrine disruptor, has high teratogenicity to embryos of amphibian (Xenopus tropicalis). An amphibian metamorphosis assay (AMA) and a complete AMA (CAMA) were conducted for TBT. In AMA, the body weight, the snout-to-vent length and the hind limb length of X. laevis tadpoles were decreased in tributyltin chloride (TBTCl; 12.5–200 ng/L) treatment groups after 7 days exposure. TBT greatly retarded the development of tadpoles, decreased the number of follicle and induced thyroid follicle cell hyperplasia after 19 days exposure. In CAMA, 10 and 100 ng/L TBTCl led to various malformations of gonad, including intersex, segmental aplasia and multiple ovary cavities of X. laevis following exposure from stages 46 to stage 66. The sex ratio was male-biased in TBT treatment groups. These results suggest that TBT delayed the metamorphosis, inhibited the growth of tadpoles and disrupted the gonadal differentiation of X. laevis at environmentally relevant concentrations.