In vitro and in vivo effects of Roundup on the ovarian growth of the estuarine crab Neohelice granulata

CANOSA, IVANA S.; SILVEYRA, GABRIELA R.; MEDESANI, DANIEL A.; RODRIGUEZ, ENRIQUE M

Buenos Aires

Congreso; SETAC Latin America 11th Biennial Meeting; 2015

SETAC Latinoamerica

The effectof Roundup Ultramax® (R-UP), one of the main glyphosate formulationcurrently used, was evaluated in vitroon ovarian pieces of the estuarine crab Neohelicegranulata, during the pre-reproductive period. Small pieces were incubatedfor 24 h in Medium199, inside a culture chamber held at 27 ºC and 5% CO2.Experimental groups comprised a control (vehicle) and three concentrations ofR-UP: R-UP1 (0,01 mg/L of glyphosate as active principle), R-UP2 (0,1 mg/L) andR-UP3 (1 mg/L). Twelve females were used, each one providing biologicalmaterial to every treatment (repeated measure design). Tritiated leucine wasadded to each 2 mL-well, in order to estimate the incorporation of this amino acidto de novo synthesis of ovarian  proteins (i.e., mainly vitellins). At the end of the assay, ovarian pieces werehomogeneized, centrifugated and filtered, to finally counting the CPM in theacid precipitable fraction. A significant (p<0.05) lower incorporation ofleucine was observed in the ovarian pieces exposed to R-UP3, with respect tocontrol. By the same time, adult females of N.granulata (N=60) were exposed for 84 d to the same concentrations assayed in vitro. Each female was isolated in aglass recipient filled with 400 mL of artificial saline water at 12 g/L, undercontinuous aeration, temperature of 22 ºC and photoperiod 14:10 (L:D). At theend of the assay, ovaries of all females were dissected, fixed in Bouinsolution and histologically processed. For each animal, all oocyte types(pre-vitellogenic, intermediate and vitellogenic and reabsorbed) were countedin three independent section of the ovary. Statistical analysis showed asignificant (p<0.05) higher proportion of reabsorbed vitellogenic oocytes inthe R-UP3 treatment, compared to control. Taken together, the results obtainedboth in vitro and in vivo are indicating a deleteriouseffect of the highest concentration of R-UP on the ovarian growth of N. granulata, also suggesting that sucheffect of is directly exerted on the ovary.