First study on response of astrocytes in alevines of red-bellied pacu (Piaractus brachypomus) to subchronic exposure to chlorpyrifos and trichlorfon.

Background and Aim: Organophosphate pesticides (OPs) used in agricultural production pose environmental and public health risks whenever non-target organisms are exposed to them. Oxon-type OPs, such as trichlorfon (TCF) and chlorpyrifos (CPF), are frequently used in Colombia and have been detected in water bodies in the vicinity of croplands; however, their effect on aquatic organisms, especially fish, is largely unknown. The neurotoxicity of OPs includes inhibition of esterase enzymes, neuronal damage, and increased glial reactivity. This study aimed to assess the astrocytic response in the brain tissue of juvenile red-bellied pacu (Piaractus brachypomus) exposed to TCF and CPF. Materials and Methods: A 25-day subchronic assay was conducted in which juvenile red-bellied pacu were exposed to CPF and TCF. After 25 days of exposure, the fish were killed and brain samples were collected and processed for immunohistochemistry to assess the morphology and reactivity of astrocytes; glial acidic fibrillary protein was used as a biomarker. Results: The brain samples from animals under subchronic exposure to OPs for 25 days showed higher cellular density as well as changes in astrocyte phenotype characterized by shortening of cytoplasmic projections, hypertrophy, and ameboid morphology compared to those from nonexposed animals. Similarly, astrocyte hyperreactivity was detected in the optic tectum and medial longitudinal fasciculus of the exposed group. Conclusion: Immunoreactivity of brain glial cells under subchronic exposure to OPs measured through immunohistochemical tests as well as OPs-induced neuropathology may be useful as a biomarker for monitoring environmental pollution. The results also indicate that P. brachypomus is a suitable biomonitoring model for studying neurotoxicological and neurodegenerative diseases.

Molecular characterization of HEPCIDIN-1 (HAMP1) gene in red-bellied pacu (Piaractus brachypomus).

Hepcidins are cysteine-rich peptides, which participate in iron metabolism regulation, the inflammatory and antimicrobial response. This study characterizes the hepcidin-1 (HAMP1) gene, its transcript expression in different tissues, as well as its regulation in a model of brain injury in Piaractus brachypomus. Bioinformatic analysis was carried out to determine conserved domains, glycosylation sites and protein structure of HAMP1, and probability that HAMP1 corresponds to an antimicrobial peptide (AMP). Relative gene expression of the P. brachypomus HAMP1 gene was determined by qPCR from cDNA of several tissues, a brain injury model, an organophosphate sublethal toxicity model and anesthetic experiment using the 2-ΔΔCt method. HAMP1 ORF encodes for a 91 aa pre-prohepcidin conformed for a prodomain with 42 aa and mature peptide of 25 aa. Mature domain was determined as an AMP. HAMP1 transcript is expressed in all the tissues, being higher in the spleen and liver. HAMP1 mRNA level was upregulated in the brain injury group, as well as in the olfactory bulb, optic chiasm and telencephalon of red-bellied pacu brain exposed to an organophosphate. In anesthetic experiment, HAMP1 mRNA level was upregulated in the liver and gills. HAMP1 gene of P. brachypomus may be involved in the inflammatory, antimicrobial, hypoxia and stress oxidative response.

Molecular characterization of myelin basic protein a (mbpa) gene from red-bellied pacu (Piaractus brachypomus).

BACKGROUND: Myelin basic protein (MBP) is one of the most important structural components of the myelin sheaths in both central and peripheral nervous systems. MBP has several functions including organization of the myelin membranes, reorganization of the cytoskeleton during the myelination process, and interaction with the SH3 domain in signaling pathways. Likewise, MBP has been proposed as a marker of demyelination in traumatic brain injury and chemical exposure. METHODS: The aim of this study was to molecularly characterize the myelin basic protein a (mbpa) gene from the Colombian native fish, red-bellied pacu, Piaractus brachypomus. Bioinformatic tools were used to identify the phylogenetic relationships, physicochemical characteristics, exons, intrinsically disordered regions, and conserved domains of the protein. Gene expression was assessed by qPCR in three models corresponding to sublethal chlorpyrifos exposure, acute brain injury, and anesthesia experiments. RESULTS: mbpa complete open reading frame was identified with 414 nucleotides distributed in 7 exons that encode 137 amino acids. MBPa was recognized as belonging to the myelin basic protein family, closely related with orthologous proteins, and two intrinsically disordered regions were established within the sequence. Gene expression of mbpa was upregulated in the optic chiasm of the chlorpyrifos exposed fish in contrast to the control group. CONCLUSIONS: The physicochemical computed features agree with the biological functions of MBP, and basal gene expression was according to the anatomical distribution in the tissues analyzed. This study is the first molecular characterization of mbpa from the native species Piaractus brachypomus.

Sex steroid hormones as neuroprotective elements in ischemia models.

Among sex steroid hormones, progesterone and estradiol have a wide diversity of physiological activities that target the nervous system. Not only are they carried by the blood stream, but also they are locally synthesized in the brain and for this reason, estradiol and progesterone are considered 'neurosteroids'. The physiological actions of both hormones range from brain development and neurotransmission to aging, illustrating the importance of a deep understanding of their mechanisms of action. In this review, we summarize key roles that estradiol and progesterone play in the brain. As numerous reports have confirmed a substantial neuroprotective role for estradiol in models of neurodegenerative disease, we focus this review on traumatic brain injury and stroke models. We describe updated data from receptor and signaling events triggered by both hormones, with an emphasis on the mechanisms that have been reported as 'rapid' or 'cytoplasmic actions'. Data showing the therapeutic effects of the hormones, used alone or in combination, are also summarized, with a focus on rodent models of middle cerebral artery occlusion (MCAO). Finally, we draw attention to evidence that neuroprotection by both hormones might be due to a combination of 'cytoplasmic' and 'nuclear' signaling.

Toxicity of trichlorfon exposure using an experimentally induced model with Silver Pacu (Piaractus brachypomus) / Efectos toxicológicos generados por la exposición a triclorfón en un modelo inducido experimentalmente en cachama blanca (Piaractus brachypomus) / Efeitos toxicológicos no peixe Pirapitinga (Piaractus brachypomus) gerado pela exposição a triclorfon em um modelo induzido experimentalmente

The trichlorfon (TCF) is an organophosphate insecticide widely used in agricultural systems and livestock production. However, its indiscriminate use has generated serious problems affecting organisms in aquatic ecosystems that are not direct targets of the chemical. Therefore, the aim of this study was to assess acute toxicity (LC50) and chronic subletal of TCF in cachama blanca fingerlings (Piaractus brachypomus). Acute toxicity was evaluated at 96 hours, using four concentrations of TCF (0.01, 0.1, 0.25, 0.5 mg/L) and a control group, each group consisted of 8 animals (2.5 ± 0.5 g). The LC50 value calculated using the Trimmed Spearman-Karber software was 0.18 mg/L (p<0,05). The evaluation of the effect of chronic exposure to sublethal TCF on the immune response of P. brachypomus was performed by testing respiratory burst, showing significant increase in animals exposed to 0.029 mg / L during the first 10 days of exposure compared to control. Likewise, the plasma bactericidal capacity was evaluated in TCF exposed fish using Aeromonas hydrophila, finding greater of colony forming units on day 20 and 30 of exposure to the concentration 0.018 mg/L as opposed to the control. In addition, a histopathological study was performed, finding gill lesions compatible with epithelial cell hypertrophy and lamellar detachment in fish exposed to 0,05 mg/L on day 30 of exposure. This work showed the cachama blanca as a biomarker in contaminations associated with these xenobiotics.

El Triclorfón (TCF) es un insecticida organofosforado ampliamente utilizado en sistemas de producción agrícola y pecuario; sin embargo, su uso indiscriminado ha generado serios problemas en ecosistemas acuáticos afectando organismos que no son blanco directo del compuesto químico. Por este motivo, el presente estudio tuvo como objetivos evaluar la toxicidad aguda (CL50) y crónica subletal del TCF en alevinos de cachama blanca (Piaractus brachypomus). La toxicidad aguda se evaluó a 96 horas, empleando cuatro concentraciones de TCF (0,01, 0,1, 0,25, 0,5 mg/L) y un grupo control, cada grupo consto de 8 animales (2,5 ± 0,5 g). El valor de CL50 calculado mediante el software Trimmed Spearman-Karber fue 0,18 mg/L (p<0,05). La evaluación del efecto de la exposición crónica subletal a TCF sobre la respuesta inmune de P. brachypomus se realizó mediante el ensayo de explosión respiratoria, evidenciándose aumento significativo en animales sometidos a 0,029 mg/L durante los primeros 10 días de exposición en comparación con el control. Asimismo, se evalúo la capacidad bactericida del plasma de los peces expuestos a TCF frente Aeromonas hydrophila, presentándose mayor conteo de unidades formadoras de colonia para el día 20 y 30 de exposición a la concentración de 0,018 mg/L a diferencia del control. Aunado a esto, se realizó estudio histopatológico, hallándose lesiones branquiales compatibles con hipertrofia de células epiteliales y desprendimiento lamelar en los peces expuestos a 0,05 mg/L para el día 30 de exposición. Este trabajo demuestra que la cachama blanca se constituye en un bioindicador de contaminación asociado a estos xenobióticos.

O triclorfom (TCF) é um inseticida organofosforado amplamente utilizado em sistemas de produção agrícola e pecuária; no entanto, seu uso indiscriminado tem levado a sérios problemas que afetam os organismos nos ecossistemas aquáticos que não são alvos diretos da substância química. Portanto, o presente estudo teve como objetivo avaliar a toxicidade aguda (CL50) e crônica subletal de TCF em alevinos de cachama blanca (Piaractus brachypomus). A toxicidade aguda foi avaliada em 96 horas, usando quatro concentrações de TCF, com um grupo control, cada grupo com 8 animais (2,5 ± 0,5 g). O valor de CL50 calculado com o software de Spearman-Karber foi de 0,18 mg/L (p<0,05). A avaliação do efeito da exposição crônica a concentrações subletais de TCF na resposta imune de P. brachypomus foi realizada pelo teste de explosão respiratória, mostrando um aumento significativo nesta, em animais expostos a 0,029 mg/L, durante os primeiros 10 dias de exposição. Também, a capacidade bactericida do plasma dos peixes expostos a TCF foi avaliada frente a Aeromonas hydrophila, os quais apresentaraõ maior unidades formadoras de colônias no dia 20 e 30 de exposição a concentração de 0,018 mg/L, ao contrario do controle. Alem disso, um estudo histopatológico foi realizado, emcontrando lesões de brânquia compatíveis com hipertrofia e desprendimento de células epiteliais nas lamelas de peixes expostos a 0,05 mg/L no dia 30 de exposição. este trabalho mostra ao cachama blanca como um biomarcador de contaminação asociado com estes xenobióticos.