Supplementary Materials

Microplastic particles cause intestinal damage and other adverse

effects in zebrafish Danio rerio and nematode Caenorhabditis

elegans

Lili Leia, Siyu Wua, Shibo Lua, Mengting Liua, Yang Songa, Zhenhuan Fua, Huahong Shib, Kathleen M. Raley-Susmanc, Defu Hea, d*

a Lab of Toxicology, School of Ecological and Environmental Sciences, East China Normal University, 500# DongChuan RD, Shanghai, 200241, Chinab State Key Laboratory of Estuarine and Coastal Research, East China Normal University, Shanghai 200062, China c Department of Biology, Vassar College, Poughkeepsie, NY 12604, USAd Shanghai Key Laboratory for Urban Ecological Processes and Eco-Restoration, East China Normal University, Shanghai, 200241, China

* Corresponding author: Defu He, Lab of Toxicology, School of Ecological and Environmental Sciences, East China Normal University, 500# DongChuan RD, Shanghai, 200241, China. Tel: +86 189 1786 4019; E-mail address: dfhe@des.ecnu.edu.cn

Supplementary Materials include 2 tables and 3 figures in Page S2-S6:

Table S1. Properties of five types of microplastic particles in the present study. Page S2Table S2. Scores and occurrence rates of intestinal damages in zebrafish after exposed to five types of microplastic particles. Page S3Fig. S1. FTIR spectroscopy for five types of microplastics in the present study. Page S4Fig. S2. Scores for intestinal fold disruption (a) and enterocyte damages (b) in D. rerio after microplastics exposure. Page S5Fig. S3. Representative photomicrographs of the gill (a), liver (c) and kidney (e) from unexposed Danio rerio. Right panel is the images of gill (b), liver (d), and kidney (f) from D. rerio exposed to microplastic particles of PA, PE, PP, PVC or PS for 10 days. Bar = 200 μm. Page S6

S1

Table S1 Properties of five types of microplastic particles in the present study

Chemical names Abbr. CAS numberMolecular

formula

Structural

formulaReagent company

Polyamide PA 63428-83-1 C6H3N3O7 Sinopharm Chemical

Polyethylene PE 9002-88-4 [C2H4]n SIGMA-ALDRICH

Polypropylene PP 9003-07-0 [C3H6]n SIGMA-ALDRICH

Poly(vinyl chloride) PVC 9002-86-2 [C2H3Cl]n SIGMA-ALDRICH

Polystyrene PS 9003-53-6 [C8H8]n Aladdin

S2

Table S2 Score values and occurrence rates of intestinal damages in zebrafish after exposed to five types of microplastic particles

Control PA PE PP PVC PS0.1 PS1.0 PS5.0

Damage

scores0.1±0.1 1.2±0.2*** 1.3±0.3*** 2.0±0.3*** 1.4±0.6*** 0.1±0.1 0.1±0.1 0.1±0.1

Occurrenc

e rates6.7% 80.0% 73.3% 86.7% 80.0% 6.7% 6.7% 13.3%

Note: Zebrafish were exposed to 1.0 mg/L microplastic particles. Intestine damages (inflammatory or broken) were

assessed by assigning a score value ranging from 0 to 4 (0 normal; 1 slight; 2 moderate; 3 pronounced; 4 severe

damages) in terms of the semi-quantitative method. Occurrence rates of intestinal damages were calculated as the

number of damage individual in total number of each group. *** p < 0.001, compared to the control. Data of

damage scores are means ± SE (n=15).

S3

Fig. S1. FTIR spectroscopy for five types of microplastics in the present study.

S4

Fig. S2. Scores for intestinal fold disruption (a) and enterocyte damages (b) in D. rerio after microplastics exposure. Zebrafish were exposed to 1.0 mg/L microplastic particles for 10 days. a: intestinal fold architecture were assessed by assigning score values ranging from 0 to 3 (0 normal, 1 slight disruption, 2 moderate disruption characterized by decreased intestinal fold and villi, 3 severe disruption). b: enterocyte damages were assessed by assigning scores ranging from 0 to 3 (0 normal, 1 slight, 2 moderate damages characterized by splitting of enterocytes, 3 severe damages). ***p < 0.001, compared to the control. Data are means ± SEM (n=15).

S5

Fig. S3. Representative photomicrographs of the gill (a), liver (c) and kidney (e) from unexposed Danio rerio. Right panel is the images of gill (b), liver (d), and kidney (f) from D. rerio exposed to microplastic particles of PA, PE, PP, PVC or PS for 10 days. Bar = 200 μm.

S6