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Page 6 of 18 Monaco et al. J Environ Expo Assess 2024;3:18 https://dx.doi.org/10.20517/jeea.2024.10

Table 1. Small and large intestinal weight and length and organ weights normalized by BW of piglets supplemented orally with 0
(CON), 20 (DEHP20), or 200 (DEHP200) mg DEHP/kg BW/day
Treatment groups
CON DEHP20 DEHP200 P-value
Intestinal length (cm/kg) 116 ± 2.5 118 ± 2.6 116 ± 2.0 0.74
Intestinal weight (g/kg) 34 ± 0.9 34 ± 1.2 34 ± 1.4 0.98
Cecum + colon (g/kg) 23 ± 1.1 22 ± 0.9 22 ± 0.7 0.53
Brain (g/kg) 6.2 ± 0.1 6.0 ± 0.1 6.3 ± 0.2 0.34
Kidneys (g/kg) 7.5 ± 0.3 7.7 ± 0.3 8.0 ± 0.5 0.59
Spleen (g/kg) 3.5 ± 0.5 4.2 ± 0.7 3.5 ± 0.6 0.68
Heart (g/kg) 6.1 ± 0.2 6.0 ± 0.2 6.7 ± 0.3 0.06
Liver (g/kg) 33 ± 0.9 31 ± 0.7 32 ± 0.7 0.20
Lung (g/kg) 13 ± 0.3 13 ± 0.5 13 ± 0.5 0.63

Data are expressed as mean ± SEM; n = 8 per group. DEHP: Di-(2-ethylhexyl) phthalate; BW: body weight; SEM: standard error of the means.

Figure 1. BW of piglets fed formula containing 0 (CON), 20 (DEHP20) or 200 (DEHP200) mg DEHP/kg BW/day from postnatal days 2
to 23. Values are means ± SEM. There were no significant differences in weight gain (P > 0.05). DEHP: Di-(2-ethylhexyl) phthalate;
BW: body weight; SEM: standard error of the means.

Urinary metabolites
Four different DEHP metabolites were detected in urine [Figure 2]. DEHP metabolite excretion in the urine
was positively correlated with DEHP dose. MEHP, MEHHP, MEOHP, and MECPP concentrations in
DEHP20 and DEHP200 were 6.6-, 7.5-, 6.3-, and 6.3-fold, and 51-, 115-, 47-, and 38-fold higher than CON,
respectively, while DEHP200 was 7.7-, 15.2-, 7.6-, and 6.1-fold higher than DEHP-20. MEOHP was the most
abundant DEHP metabolite found in urine, followed by MEHHP and MEHP.

Intestinal histomorphology
Morphological measurements of small intestine villi and crypts and AC cuffs are presented in Table 2.
Exposure to DEHP did not affect intestinal morphology in the ileum or AC. A statistical trend (P < 0.07)
was observed toward higher crypt depth in the duodenum of DEHP200 compared to CON and DEHP20.
Jejunal villus length was significantly shorter in piglets fed the high dose of DEHP, while villus area was
lower in DEHP200 than DEHP20. Jejunum villus area between DEHP and CON tended (P = 0.07) to differ.
Jejunal crypt depth was highest in DEHP200, intermediate in CON, and lowest in DEHP20, while crypt area
was lower in DEHP20 than CON and DEHP200.