Effects of the Herbicide Atrazine on Crustacean Reproduction: Mini-Review

Atrazine, one of the most intensively applied herbicides worldwide, is commonly found in several water bodies, affecting the associated fauna. Autochthon crustacean species have been relatively less studied, compared to vertebrate species, particularly concerning reproductive success. In this mini-review, we summarize the relevant information about the effects of atrazine exposure on the main reproductive aspects of crustaceans. One of these effects is related to the inhibition of ovarian growth. In this respect, a diminished vitellogenin content was found in the ovary of crabs exposed to atrazine during the entire period of ovarian growth, in correlation with a reduced oocyte size and a delay of ovarian maturation. Similar results were observed in crayfish. Atrazine was also able to affect the reproductive process, acting as an endocrine disruptor. In this sense, this herbicide was suspected to affect the secretion of some neurohormones involved in the gonadal growth, as well as to alter the circulating levels of steroid hormones which promote the synthesis of vitellogenin for ovarian growth. Moreover, atrazine induced sexual differentiation in juvenile crayfish toward a higher proportion of females, while it produced an increment of males in daphnids. Another aspect affected by this herbicide was the reduction of offspring production, as well as several embryonic abnormalities; genotoxic effects have been also reported in crayfish. Finally, some metabolic imbalances, such as reduction in energy reserves, have been observed in some species, together with oxidative stress and histopathological effects.Fil: Silveyra, Gabriela Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Medesani, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Rodriguez, Enrique Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

Interference of an atrazine commercial formulation with the endocrine control of ovarian growth exerted by the eyestalks

Atrazine is currently one of the most used herbicides worldwide. We tested the possible effect of the widely used herbicide atrazine on the endocrine control of ovarian growth exerted by the neurohormones secreted at the eyestalk of the estuarine crab Neohelice granulata. For this, both in vivo and in vitro assays were carried out. The in vivo assay comprised the exposure for 1 month to 3 mg/L of a commercial formulation containing 90% of atrazine as active ingredient (Gesaprim 90 WDG®, Syngenta) on three categories of females: intact, ablated of one eyestalk, and ablated of both eyestalks. At the end of the assay, only the intact females showed a significant (p < 0.05) decrease in both content of vitellogenic ovarian proteins and proportion of vitellogenic oocytes, compared to a concurrent control. The results of the in vitro incubation of ovarian pieces with the eventual addition to the incubation medium of eyestalk tissue and/or atrazine at 3 mg/L showed a significant (p < 0.05) decrease in the proportion of vitellogenic oocytes only when atrazine and eyestalk tissue were added. Taken together, these results strongly suggest that the assayed atrazine formulation may act as an endocrine disruptor at the eyestalk level, by altering the normal secretion of some eyestalk hormone, therefore inhibiting ovarian growth.Fil: Silveyra, Gabriela Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Canosa, Ivana Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Zanitti, Marina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Rodriguez, Enrique Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Medesani, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

Early high-titer plasma therapy to prevent severe Covid-19 in older adults

BACKGROUND: Therapies to interrupt the progression of early coronavirus disease 2019 (Covid-19) remain elusive. Among them, convalescent plasma administered to hospitalized patients has been unsuccessful, perhaps because antibodies should be administered earlier in the course of illness. METHODS We conducted a randomized, double-blind, placebo-controlled trial of convalescent plasma with high IgG titers against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in older adult patients within 72 hours after the onset of mild Covid-19 symptoms. The primary end point was severe respiratory disease, defined as a respiratory rate of 30 breaths per minute or more, an oxygen saturation of less than 93% while the patient was breathing ambient air, or both. The trial was stopped early at 76% of its projected sample size because cases of Covid-19 in the trial region decreased considerably and steady enrollment of trial patients became virtually impossible. RESULTS A total of 160 patients underwent randomization. In the intention-to-treat population, severe respiratory disease developed in 13 of 80 patients (16%) who received convalescent plasma and 25 of 80 patients (31%) who received placebo (relative risk, 0.52; 95% confidence interval [CI], 0.29 to 0.94; P = 0.03), with a relative risk reduction of 48%. A modified intention-to-treat analysis that excluded 6 patients who had a primary end-point event before infusion of convalescent plasma or placebo showed a larger effect size (relative risk, 0.40; 95% CI, 0.20 to 0.81). No solicited adverse events were observed. CONCLUSIONS Early administration of high-titer convalescent plasma against SARS-CoV-2 to mildly ill infected older adults reduced the progression of Covid-19. (Funded by the Bill and Melinda Gates Foundation and the Fundación INFANT Pandemic Fund; Dirección de Sangre y Medicina Transfusional del Ministerio de Salud number, PAEPCC19, Plataforma de Registro Informatizado de Investigaciones en Salud number, 1421, and ClinicalTrials.gov number, NCT04479163.).Fil: Libster, Romina Paula. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Pérez Marc, Gonzalo. Hospital Militar Central, Buenos Aires; ArgentinaFil: Wappner, Diego. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Coviello, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Bianchi, Alejandra. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Braem, Virginia. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Esteban, Ignacio. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Caballero, Mauricio Tomás. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Wood, Cristian. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Berrueta, Mabel. Hospital Militar Central; ArgentinaFil: Rondan, Aníbal. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Lescano, Gabriela Mariel. Hospital Dr. Carlos Bocalandro; ArgentinaFil: Cruz, Pablo. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Ritou, Yvonne. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Fernández Viña, Valeria Silvina. Hospital Simplemente Evita; ArgentinaFil: Álvarez Paggi, Damián Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Esperante, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Ferreti, Adrián. Hospital Dr. Carlos Bocalandro; ArgentinaFil: Ofman, Gaston. University of Oklahoma; Estados UnidosFil: Ciganda, Álvaro. Gobierno de la Provincia de Buenos Aires. Hospital Interzonal Especializado de Agudos y Cronicos San Juan de Dios.; ArgentinaFil: Rodriguez, Rocío. Hospital Simplemente Evita; ArgentinaFil: Lantos, Jorge. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Valentini, Ricardo. No especifíca;Fil: Itcovici, Nicolás. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Hintze, Alejandra. No especifíca;Fil: Oyarvide, M. Laura. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Etchegaray, Candela. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Neira, Alejandra. Instituto de Efectividad Clínica y Sanitaria; ArgentinaFil: Name, Ivonne. Instituto de Efectividad Clínica y Sanitaria; ArgentinaFil: Alfonso, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Swiss Medical Group; ArgentinaFil: López Castelo, Rocío. Centro de Educación Médica e Investigaciones Clínicas "Norberto Quirno"; ArgentinaFil: Caruso, Gisela. Hospital Militar Central; ArgentinaFil: Rapelius, Sofía. Hospital Militar Central; ArgentinaFil: Alvez, Fernando. Hospital Militar Central; ArgentinaFil: Etchenique, Federico. Hospital Militar Central; ArgentinaFil: Dimase, Federico. Hospital Militar Central; ArgentinaFil: Alvarez, Darío. Hospital Militar Central; ArgentinaFil: Aranda, Sofía S.. Hospital Militar Central; ArgentinaFil: Sánchez Yanotti, Clara Inés. Hospital Militar Central; ArgentinaFil: De Luca, Julián. Hospital Militar Central; ArgentinaFil: Jares Baglivo, Sofía. Hospital Militar Central; ArgentinaFil: Laudanno, Sofía. Fundación Hematológica Sarmiento; ArgentinaFil: Nowogrodzki, Florencia. Swiss Medical Group; ArgentinaFil: Larrea, Ramiro. Hospital Municipal San Isidro; ArgentinaFil: Silveyra, María. Hospital Militar Central; ArgentinaFil: Leberzstein, Gabriel. No especifíca;Fil: Debonis, Alejandra. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Molinos, Juan. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: González, Miguel. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Perez, Eduardo. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Kreplak, Nicolás. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Pastor Argüello, Susana. Fundación para la Investigación en Infectología Infantil; ArgentinaFil: Gibbons, Luz. Hospital Municipal de San Isidro; ArgentinaFil: Althabe, Fernando. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Efectividad Clínica y Sanitaria; ArgentinaFil: Bergel, Eduardo. Sanatorio Sagrado Corazón; ArgentinaFil: Polack, Fernando Pedro. Provincia de Buenos Aires. Ministerio de Salud; Argentin

Effects of atrazine on ovarian growth and its endocrine control in decapod crustaceans

El herbicida atrazina es uno de los principales plaguicidas aplicados de manera intensiva a nivel mundial. El objetivo del presente trabajo fue evaluar el efecto de atrazina sobre la reproducción y su control endocrino en crustáceos decápodos. Para esto, se realizaron ensayos in vivo con hembras del cangrejo Neohelice granulata, que fueron expuestas de manera crónica a tres concentraciones subletales (0,05; 0,5 y 5 mg/L) de una formulación comercial o bien de una formulación experimental de atrazina, durante tres meses. Se observó un descenso en el contenido total de proteínas vitelogénicas en el ovario, un menor tamaño de oocitos pre-vitelogénicos y vitelogénicos, y un descenso en la proporción de oocitos vitelogénicos en los animales expuestos a 5 mg/L del formulado comercial, evidenciando un retraso en el crecimiento ovárico. Estos resultados no fueron observados en las hembras expuestas a la formulación experimental, revelando una toxicidad diferencial entre ambos formulados. Además, se observó un descenso en el contenido de glucógeno y proteínas en el músculo de las hembras expuestas a 5 mg/L, posiblemente como respuesta compensatoria al efecto estresante inespecífico del herbicida. Por otro lado, en animales ablacionados de ambos pedúnculos oculares y expuestos a 5 mg/L del formulado comercial, no se observaron cambios en ninguna de las mismas variables medidas en las hembras intactas, indicando que el retraso en el crecimiento ovárico podría deberse a una interferencia del herbicida con la regulación del proceso de vitelogénesis dada por neurohormonas pedunculares. Este mecanismo de acción del herbicida se corroboró mediante ensayos in vitro, incubando ovarios con pedúnculos oculares en presencia o ausencia de atrazina; el análisis histológico del ovario evidenció que el formulado comercial estaría actuando como un desorganizador endocrino, posiblemente estimulando la secreción desde los pedúnculos oculares de la neurohormona inhibidora del desarrollo gonadal, afectando así negativamente el crecimiento ovárico. Varios de los resultados obtenidos en N. granulata fueron también observados en la langosta Procambarus clarkii. En particular, se observó un descenso en la expresión relativa de vitelogenina (Vg) ovárica y hepatopancreática y un menor contenido de Vg en el ovario de hembras expuestas a 5 mg/L del formulado comercial, así como una disminución en el tamaño de los oocitos en hembras expuestas a 1 y 5 mg/L; esta inhibición del crecimiento ovárico fue similar a la observada en N. granulata. Asimismo, se determinaron en P. clarkii los niveles de algunos esteroides sexuales y marcadores de estrés metabólico y oxidativo; los niveles circulantes de estradiol se vieron aumentados en los animales expuestos, demostrando que el descenso en la expresión relativa y en el contenido de Vg ovárica no estaría asociado con una menor secreción de esta hormona. Las hembras expuestas a atrazina mostraron un incremento de lactato y glutatión circulante, indicando un mayor esfuerzo metabólico y una cierta activación de las defensas antioxidantes, en respuesta al estrés generado por el herbicida. En su conjunto, los resultados obtenidos evidencian que el formulado comercial de atrazina utilizado altera la dinámica normal del crecimiento gonadal, en hembras de dos especies de crustáceos y, en consecuencia, podría afectar de manera directa a sus poblaciones, poniendo de manifiesto los efectos nocivos de este tipo de sustancias para el ambiente y los seres vivos que habitan en él.The herbicide atrazine constitutes one of the most widely used pesticides in the world. The present study was aimed at evaluating the effect of atrazine on reproductive function and its endocrine control, in two crustacean decapod species. For this, we ran in vivo assays, in which females of the crab Neohelice granulata were chronically exposed to three subletal concentrations (0.05, 0.5 and 5 mg/L) of either an experimental formulation or a commercial formulation of atrazine, for three months. We observed a decrease in total content of the ovarian protein vitelogenin (Vg) and a significant decrease in pre-vitellogenic and vitellogenic oocyte size, together with a lower proportion of vitellogenic oocytes in animals exposed to 5 mg/L of the commercial formulation, showing a delay in ovarian maturation. These results were not observed with the experimental formulation, indicating a differential toxicity between both formulations. In addition, we observed a decrease in glycogen and protein levels in muscle of females exposed to 5 mg/L, likely due to a compensatory effect of the stress induced by the exposure to the herbicide. On the other hand, in animals that were ablated of both eyestalks before exposing them to 5 mg/L of atrazine, no changes were observed in the same variables measured in intact females, indicating that the delay in ovarian maturation may be due to an interference of the herbicide on the eyestalk hormonal regulation of vitellogenesis. This mechanism of action of the herbicide was corroborated by means of in vitro assays, incubating ovarian pieces with eyestalks in the presence or absence of atrazine. Histological analysis of ovaries revealed that the commercial formulation may be acting as an endocrine disruptor, potentially inducing the secretion of gonad inhibiting hormone from the eyestalks, therefore reducing ovarian growth. Several of effects observed in N. granulata were also seen in the freshwater crayfish Procambarus clarkii. Particularly a reduction in the relative expression of both ovarian and hepatopancreatic Vg and a lower ovarian Vg content in females exposed to 5 mg/L of commercial atrazine were observed, as well as a reduction in the oocyte size, in females exposed to both 1 and 5 mg/L of the commercial formulation; this inhibition was similar to that observed in N. granulata. We also measured some steroid hormone levels and oxidative and metabolic stress markers. We found that circulating levels of estradiol were increased in exposed females, indicating that the reduction in the relative expression and content of ovarian Vg may not be associated with a reduction in estradiol secretion. Females exposed to atrazine showed increased levels of circulating lactate and glutathione, indicating a higher metabolic effort and some activation of antioxidant defenses, in response to the stress induced by the herbicide. Taken together, the observed results demonstrate that the commercial formulation of atrazine alters the normal ovarian growth in females of the two crustacean species, and may directly affect their natural populations, providing evidence of the harmful effects of these chemicals to the environment and the species living there.Fil: Silveyra, Gabriela Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

Biomarkers for Bronchopulmonary Dysplasia in the Preterm Infant

Bronchopulmonary dysplasia (BPD) is a chronic inflammatory lung disease of very-low-birth-weight (VLBW) preterm infants, associated with arrested lung development and a need for supplemental oxygen. Over the past few decades, the incidence of BPD has significantly raised as a result of improved survival of VLBW infants requiring mechanical ventilation. While early disease detection is critical to prevent chronic lung remodeling and complications later in life, BPD is often difficult to diagnose and prevent due to the lack of good biomarkers for identification of infants at risk, and overlapping symptoms with other diseases, such as pulmonary hypertension (PH). Due to the current lack of effective treatment available for BPD and PH, research is currently focused on primary prevention strategies, and identification of biomarkers for early diagnosis, that could also represent potential therapeutic targets. In addition, novel histopathological, biochemical, and molecular factors have been identified in the lung tissue and in biological fluids of BPD and PH patients that could associate with the disease phenotype. In this review, we provide an overview of biomarkers for pediatric BPD and PH that have been identified in clinical studies using various biological fluids. We also present a brief summary of the information available on current strategies and guidelines to prevent and diagnose BPD and PH, as well as their pathophysiology, risk factors, and experimental therapies currently available.Fil: Rivera, Lidys. State University of Pennsylvania; Estados UnidosFil: Siddaiah, Roopa. State University of Pennsylvania; Estados UnidosFil: Oji Mmuo, Christiana. State University of Pennsylvania; Estados UnidosFil: Silveyra, Gabriela Romina. State University of Pennsylvania; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Silveyra, Patricia. State University of Pennsylvania; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Effects of atrazine on vitellogenesis, steroid levels and lipid peroxidation, in female red swamp crayfish Procambarus clarkii

Atrazine, a widely use herbicide, has been classified as a potential endocrine disruptor, especially for freshwater species. In this study, we tested the hypothesis that atrazine can affect reproduction in crayfish through dysregulation of vitellogenin expression and hormone synthesis. Adult female crayfish (Procambarus clarkii) were exposed during one month to atrazine at concentrations of either 1 or 5 mg/L. At the end of the exposure, ovaries, hepatopancreas, and hemolymph samples were harvested for analysis of vitellogenin expression and steroid hormone levels. Ovarian tissue was also sampled for both biochemical and histological analyses. Our results show that atrazine-exposed crayfish had a lower expression of vitellogenin in the ovary and hepatopancreas, as well as smaller oocytes, and reduced vitellogenin content in the ovary. Despite these effects, circulating levels of estradiol increased in females exposed to 5 mg/L of atrazine, showing that the inhibiting effect of atrazine on vitellogenin production was not related to a lower secretion of sexual steroids. Instead, some early stimulating effects of estradiol on vitellogenesis could have occurred, particularly in the hepatopancreas. On the other hand, atrazine caused a higher metabolic effort, in terms of lactate production, presumably triggered to provide the energy needed to face the unspecific stress produced by the herbicide. Lipid peroxidation was not affected by atrazine, but glutathione levels were significantly increased.Fil: Silveyra, Gabriela Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Silveyra, Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Penn State College of Medicine; Estados UnidosFil: Vatnick, Itzick. Widener University; Estados UnidosFil: Medesani, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Rodriguez, Enrique Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

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.Fil: Silveyra, Gabriela Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Silveyra, P.. Indiana University; Estados UnidosFil: Brown, M.. Widener University; Estados UnidosFil: Poole, S.. Widener University; Estados UnidosFil: Vatnick, I.. Widener University; Estados UnidosFil: Medesani, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; ArgentinaFil: Rodriguez, Enrique Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentin

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 3 mg/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 3 mg/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.Fil: Silveyra, Gabriela Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Canosa, Ivana Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Rodriguez, Enrique Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Medesani, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

Ovarian growth impairment after chronic exposure to Roundup Ultramax® in the estuarine crab Neohelice granulata

Adult females of the estuarine crab Neohelice granulata were exposed to the glyphosate formulation Roundup Ultramax® during the entire 3-month pre-reproductive period. At the end of the assay, a significant higher increment of glycemia was noted at both glyphosate concentrations assayed (0.01 and 0.2 mg/L, acid equivalent). Although no differences were observed in the gonadosomatic index, a significantly higher proportion of reabsorbed vitellogenic oocyte was observed at the highest glyphosate concentration, together with a significant decrease of vitellogenin content in the ovary. In addition, some in vitro assays were carried out by co-incubating small pieces of ovary with or without the addition of Roundup; at both concentrations tested (same as those used in vivo), a decrease in the ovarian vitellogenin content was observed, whereas the ovarian protein synthesis was significantly inhibited by glyphosate at 0.2 mg/L in the Roundup formulation used.Fil: Canosa, Ivana Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Silveyra, Gabriela Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Avigliano, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Medesani, Daniel Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Rodriguez, Enrique Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentin

In vitro evaluation of the interference of the the herbicide glyphosate on the endocrine control of ovarian growth, by neurohomones and prostaglandins, in the estuarine crab Neohelice granulata

Adult females of the crab Neohelice granulata sampled at from Samborombón Bay (Buenos Aires Province, Argentina), were used. Several in vitro tests were carried out during the pre-reproductive period, in order to evaluate the growth of the isolated ovary in the presence of the herbicide (in its commercial formulation Roundup) and/or extracts of neuroendocrine organs or inhibitors of prostaglandin synthesis. For each assay, 12 females were used, assigning a different treatment to each portion of the ovary of the same female (randomized block design). The ovary pieces were incubated for 24 h in M199 culture medium, at 24 ºC and controlled atmosphere. At the end of each assay, the ovaries were processed either for histological analysis or to determine the levels of vitellogenin (Vg) using the ELISA technique. The results of the assays conducted with extracts of eyestalk or thoracic ganglion indicated that Roundup (RUP) addition did not significantly (p>0.05) modify the effect of such extracts on ovarian growth. However, in order to assess whether the effect of RUP on isolated ovaries is related to the hormonal signaling exerted by prostaglandins (PG), an in vitro assay with ibuprofen (IBF, inhibitor of PG synthesis) was performed. The level of Vg in the treatment with RUP 0.2 mg/L + IBF 10 mg/L was significantly lower (p0.05) modify the effect of such extracts on ovarian growth. However, in order to assess whether the effect of RUP on isolated ovaries is related to the hormonal signaling exerted by prostaglandins (PG), an in vitro assay with ibuprofen (IBF, inhibitor of PG synthesis) was performed. The level of Vg in the treatment with RUP 0.2 mg/L + IBF 10 mg/L was significantly lower (p<0.05) than that of the Control, RUP 0.2 mg/L, or IBF 10 mg groups. Although RUP per se produced a decrease (p<0.05) in Vg levels, compared to control, its combination with IBF caused a higher (p<0.05) decrease. Given the additivity of inhibitory effects observed between ibuprofen and the RUP, it is plausible to propose that the mode of action of the RUP could be due to the inhibition of the endogenous synthesis of PGs in the ovary, although the possible interference of the RUP with the PGs receptor, or the inhibition of some step in the prostaglandin hormone transduction pathway, cannot yet be ruled out.Fil: Canosa, Ivana Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Silveyra, Gabriela Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Lofrano, Juliana Leticia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Medesani, Daniel Alberto. Universidad de Buenos Aires. Facultad de Cs.exactas y Naturales. Departamento de Biodiversidad y Biologia Experimental. Laboratorio de Fisiologia de Crustáceos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; ArgentinaFil: Rodriguez, Enrique Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina. Universidad de Buenos Aires. Facultad de Cs.exactas y Naturales. Departamento de Biodiversidad y Biologia Experimental. Laboratorio de Fisiologia de Crustáceos; Argentin