Plastic waste is continuously introduced not only into marine, but also freshwater environments, where it fragments into microplastics. Organisms may be affected by the particles themselves due to ingestion and indirectly via incorporated additives such as plasticizers, since these substances have the ability to leach out of the polymer matrix. Although it has been indicated that the likelihood of additives leaching out into the gut lumen of organisms exposed to microplastics is low, studies distinguishing between the effects of the synthetic polymer itself and incorporated additives of the same polymer are scarce. Since this is obligatory for risk assessment, we analyzed the chronic effects of flexible polyvinylchloride (PVC), a widely used polymer, containing the plasticizer diisononylphthalate (DiNP) on morphology and life history of the freshwater crustacean Daphnia magna and compared these effects with the effects of rigid PVC, lacking DiNP, as well as a glass bead control. After up to 31 days of exposure, rigid PVC and glass beads did not affect body length and relative tail spine length of D. magna, whereas flexible PVC led to an increased body length and a reduced number of offspring. None of the treatments increased the mortality significantly. We were able to show that 2.67μg/L DiNP leached out of the flexible PVC into the surrounding medium using GC-MS. Yet, we were not able to measure leachate inside the gut lumen of D. magna. The effects emerged towards the end of the experiment, due to the time dependent process of leaching. Therefore, the results highlight the relevance of long-term chronic exposure experiments, especially as leaching of additives takes time. Further, our study shows the importance to distinguish between microplastics containing leachable additives and the raw polymer in ecotoxicological testing.

Thais gradata samples were collected from six locations along the southern shores of Peninsular Malaysia and analyzed for imposex incidence and organotin (butyltins (BTs), including tributyltin, dibutyltin, and monobutyltin, and phenyltins (PTs), including triphenyltin, diphenyltin, and monophenyltin concentrations), in tissue burden. On average, the BT levels were found to be higher than the levels reported a year before for other biota. In contrast, the PT compounds were found to be lower than in the previous report. The present study also classified the morphological expressions of the imposex scheme for this species into seven stages (stage 0 to stage 6) by observing the development of vas deference sequence (VDS) and penis bulk. This latest imposex scheme clearly indicates the mechanism of VDS growth, the structural changes from penis bulk to a penis with flagellum, and from a normal vaginal opening to a swollen vaginal opening. The degree of imposex was assessed using the vas deference sequence index and the percentage of females possessing the imposex characteristics. It was found that locations possessing high imposex levels also tended to show high BT levels in the snail tissue samples. However, correlation analysis did not show a significant relationship among the two parameters. A better result could be obtained if more samples and sampling locations were added in order to prove the hypothesis. The nonsignificant correlation between the shell height and any of the organotin compounds, along with no significant differences between BT levels in female and lower imposex stage samples, suggests that the BT concentration detected was a recent contamination.

The effect of aluminium on histopathological and behavioral changes in the planarian Polycelis felina (Daly.) in laboratory conditions was studied. The planarians were treated with seven concentrations of aluminium sulfate for five days and compared to three control groups of animals. Microscopical, histological and morphometric methods were used. The results showed distinguished morphological changes on the planarian body as well as behavioral changes: various depigmentations, disordered locomotion, twisting of the body parts, hardly reacting to the mechanical stimuli, body contractions, deformations and mortality. Histomorphometric analyses showed changes in the size and number of neoblasts and reticular cells. Changes in epithelial cells and segments, parenchyma and muscular layer were also noticed. Concentrations of 600 and 1100 mg/L caused most distinct changes in behavior, morphological and histological structure of planarians. The highest concentrations caused irreversible changes. Considering high LC₅₀ (1100 mg/L), the used concentrations of aluminium presented no actual environmental threat to planarian populations in nature, but showed the extent of damages in a possible concentrated aluminium environment.

Amphibians are considered to be excellent bioindicators to their morphophysiological characteristics and life cycle. In this context, the present study investigated the morphological integrity of anuran larvae collected in preserved environments in the Emas National Park, in the municipality of Mineiros (Goiás state, Brazil), and in environments representative of the agricultural matrix of the Rio Verde region, also in Goiás, where there is a long history of the use of agricultural pesticides. Samples of water from temporary ponds, permanent dams, and veredas were analyzed for the presence of pesticides and, especially atrazine (5350 μg/L), found at significantly higher concentrations in the agricultural matrix. We observed a high percentage (approximately 10%) of morphological malformations including alterations of the fins in Boana albopunctatus and Scinax fuscovarius; alteration in oral structures in B. albopunctatus, Dematonotus muelleri, Physalaemus centralis, Physalaemus cuvieri, and Leptodactylus fuscus mainly in the tadpoles collected in the agricultural environment in comparison with those from the protected area (3.5%; P < 0.0001, χ² 31.75). However, changes in the eyes, mouth, intestines, and nostrils, as well as amelia were observed only in the agricultural environment. The vast majority of the observed malformations were associated with the tail and oral disc, which suggests that these anatomical parameters may be used as sensitive morphological biomarkers. Given these findings, we reinforce that areas of agricultural land may have a deleterious effect on the morphological integrity of the tadpoles and consequently, on their development, and that these features may be used as indicators of environmental quality and health.