Multigenerational exposure to trace concentrations of DDT residues in Wistar rats: Effects on biometric development and biochemical parameters

Abstract

The Organochlorine Dichlorodiphenyltrichloroethane (DDT) and its residues, Dichlorodiphenyldichloroethane (DDD) and Dichlorodiphenyldichloroethylene (DDE), are Persistent Organic Pollutants (POPs) that bioaccumulate, persist in the environment, and magnify through the food chain. Chronic exposure is linked to oxidative stress and mitochondrial dysfunction, emphasizing the need to study its multigenerational impacts on health and development. This study investigated the effects of multigenerational exposure to DDT residues in Wistar rats. Pregnant females were provided water containing trace concentrations of p,p'-DDD (0.015 µM) and p,p'-DDE (0.006 µM) from the first day of gestation (PD0) until the end of the life cycle of two generations (F1 and F2). Biometric and biochemical evaluations were conducted at PND35 and PND105, including weight, naso-anal length, and abdominal circumference. Hepatic, renal, and adipose tissues were analyzed macro- and microscopically, along with biochemical analyses. Statistical analyses included ANOVA and generalized linear models. The hypothetical model confirmed that no significant variations occurred between generations, indicating that effects were driven by group, age, and sex differences. The analysis revealed that DDD/DDE synergism and female sex significantly influenced hepatic, renal, cerebral, and white adipose tissues. DDD/DDE exposure increased hepatic enzyme activity, reduced cerebral cholinesterase and renal antioxidants, and altered adipocyte mass. Age also influenced enzymatic activity and development, with notable differences between PND35 and PND105 in tissues and biometric indices. In conclusion, DDD/DDE exposure, particularly in females, significantly impacted hepatic, renal, cerebral, and adipose tissues. The results highlight that observed effects depend on group, age, and sex, emphasizing the risks associated with environmental contamination.

Keywords: DDD; DDE; Organochlorine; Pesticides.

Fig. 1

Experimental design. g0 – Parental generation; F1 – First generation; F2 – Second generation; PND – Postnatal days; PD0 – Pregnancy day 0; F – Females; M – Males; ACS – Mating; F – Females; M – Males. Created in BioRender. Bittencourt guimarães, A. (2025) https://BioRender.com/b09u490.

Fig. 2

Hypothetical model of tissue response influenced by group, sex, age, and generation factors. The factors considered statistically significant standardized weights supporting the proposed model were categorized as Group (CTL – 0, DDD – 1, DDE – 2, DDD/DDE – 3), Sex (M – 1, F – 2), Age (PND35 – 1, PND105 – 2), and Generation (F1 – 1, F2 – 2). Created in BioRender. Bittencourt guimarães, A. (2025) https://BioRender.com/b09u490.

Fig. 3

Macroscopic morphological changes in renal tissue and the presence of urinary lithiasis in rats exposed to residues of DDT. (A) The photo shows the right kidney (1.76 ×1.2 cm; 0.842 g) and left kidney (1.48 ×0.89 cm; 0.530 g) of an animal from the DDD group, PND35, F1 generation, male sex; (B) Bladder of an animal from the DDE group, PND105, F1 generation, male sex, presenting a calculus; (C) Calculus extracted from the bladder of an animal from the DDE group, PND105, F1 generation, male sex.

Fig. 4

Benign hepatic nodule in rats exposed to residues of DDT at PND105 in F2 generation. Figures A-C show sessile nodules measuring approximately 343 mm³ , with a translucent surface and serous-looking liquid content found in an animal from the DDD/DDE group, PND105, F2 generation, female (A-B) and in an animal from the DDE group, PND105, F2 generation, male (C). Figures D-F show the histological analysis of the hepatic nodule, with a cavity surrounded by dense fibrous connective tissue (D), intense angiogenesis, chronic plasmocytic inflammation with Russell bodies and eosinophils (E), and the presence of foamy macrophages at the lumen cavity border and mucous material (F).

Fig. 5

Principal component analysis (PCA) ordination diagrams. A) Cerebral response; B) Hepatic response; C) Renal response; D) White adipose tissue response; E) Body development. Dim.1 – Dimension 1; Dim. 2 – Dimension 2. AChE – Cerebral Cholinesterase; BuChE – Butyrylcholinesterase; CAT – Catalase; FC Total – Total food consumption; GPx – Glutathione Peroxidase; GR – Glutathione Reductase; GST – Glutathione-S-Transferase; GPT – Glutamic Pyruvic Transaminase; LI – Lee index; LPO – Lipoperoxidation; PAT – Perigonadal adipose tissue; SI – Somatic index of each organ; SOD – Superoxide Dismutase; VAT – Visceral adipose tissue; WC Total – Total water consumption; WGTIR – Weight Gain-to-Caloric Intake Ratio; WGR – Specific weight gain rate; ∆AC – Abdominal circumference gain; ∆NAL – Naso-anal length gain.

Fig. 6

Confirmatory analysis of the hypothetical model of each tissue's response to the influence of the group, sex, age, and generation factors. The statistically significant standardized weights that support the proposal were categorized as Group (CTL – 0, DDD – 1, DDE – 2, DDD/DDE – 3), Sex (M – 1, F – 2), and Age (PND35 – 1, PND105 – 2). WAT: White Adipose Tissue; BD: Body Development. Lines represent cause-and-effect relationships, and dashed lines represent associations between the effects in each tissue. *p < 0.05; * *p < 0.01; * **p < 0.001; * ** *p < 0.0001.