Oxidative stress and histopathological effects by microplastic beads, in the crayfish Procambarus clarkii, and fiddler crab Leptuca pugilator.

The present study was aimed at evaluating the in vivo effects of microplastics (MP), in terms of oxidative stress and histopathological effects, in two crustacean species: Procambarus clarkii and Leptuca pugilator. In addition, MP accumulation in the hepatopancreas (HP) of both species was also determined. Adults of both crayfish and crabs were exposed for one month to fluorescent polystyrene beads (size: 1 µm) at nominal concentrations of 1000 or 5000 particles/mL. During the exposure, animals were maintained under controlled feeding, aeration, temperature, and photoperiod conditions. At the end of the exposure, HP and hemolymph (HL) samples were harvested for analysis of oxidative damage and total antioxidant levels. Additionally, the presence of MPs in both tissues was confirmed. Significant differences with the control groups were observed in lipid peroxidation levels in HP in animals exposed to the lowest concentration in P. clarkii and to the highest concentration in L. pugilator. A marked increase in antioxidant levels was also observed in the HL at both concentrations in P. clarkii, and at the highest MPs concentration in L. pugilator. Moreover, several histopathological changes were detected in both gills and HP, including hypertrophied lamellae, lifting or collapse of gill epithelia, loss of normal shape of hepatopancreatic tubules, and epithelial atrophy in the HP tissue. We conclude that exposure to MP beads at selected concentrations results in oxidative damage, induces histopathological changes in gills and HP, and triggers an antioxidant response in two crustacean species.

Effects of atrazine on ovarian growth, in the estuarine crab Neohelice granulata.

Atrazine, a herbicide that is intensively used in Argentina, was assayed to evaluate the alteration of reproduction in a wild species of crustaceans. Adult females of the estuarine crab Neohelice granulata were exposed to formulated atrazine during the 3-month pre-reproductive period. Three atrazine concentrations (0.03, 0.3 and 3mg/L) were assayed, together with a water dilution control. At the end of the exposure period, several variables concerning the energetic status of animals were measured, such as weight gain, glycemia, and energy reserves in both muscle and hepatopancreas. The gonadosomatic index (GSI) was also determined, as well as the proportion and relative area of each oocyte type in histological sections. Besides, the total content of vitellogenin proteins (Vg) in both ovary and hepatopancreas was measured. A significant (p<0.05) decrease of glycogen content was observed in muscle, while a significant (p<0.05) lower area of both previtellogenic and vitellogenic oocytes was verified in the ovary by effect of atrazine, in correspondence with a Vg content significantly (p<0.05) diminished in the ovary and augmented in the hepatopancreas. Besides, a higher proportion of previtellogenic oocytes were seen by effect of atrazine. Taken together, these results indicate a clear reduction and delay in the ovarian growth of the studied species, during the period at which the ovary normally grows up prior to spawning. On the other hand, a decrease of Vg content was observed at 0.3 and 3mg/L of atrazine after 24-h in vitro assays carried out with ovarian explants, providing additional evidence about the inhibitory role of this herbicide on the ovarian growth. These results encourage future research on deleterious effects of atrazine on crustacean reproduction.