Toxinology in the proteomics era: a review on arachnid venom proteomics

The word venomics was coined to acknowledge the studies that use omics to investigate venom proteins and peptides. Venomics has evolved considerably over the last 20 years. The first works on scorpion or spider venomics were published in the early 2000's. Such studies relied on peptide mass fingerprinting (PMF) to characterize venom complexity. After the introduction of new mass spectrometers with higher resolution, sensitivity and mass accuracy, and the next-generation nucleotide sequencing, the complexity of data reported in research on scorpion and spider venomics increased exponentially, which allowed more comprehensive studies. In the present review article, we covered key publications on scorpion venomics and spider venomics, presenting historical grounds and implemented technologies over the last years. The literature presented in this review was selected after searching the PubMed database using the terms "(scorpion venom) AND (proteome)" for scorpion venomics, and "(spider venom) AND (proteome)" for publications on spider venomics. We presented the key aspects related to proteomics in the covered papers including, but not restricted to, the employed proteomic strategy (i.e., PMF, two-dimensional gel electrophoresis, shotgun/bottom-up and/or top-down/peptidome), and the type of mass spectrometer used. Some conclusions can be drawn from the present study. For example, the scorpion genus Tityus is the most studied concerning venomics, followed by Centruroides; whereas for spiders the studied genera were found more equally distributed. Another interesting conclusion is the lack of high throughput studies on post-translational modifications (PTMs) of scorpion and spider proteins. In our opinion, PTMs should be more studied as they can modulate the activity of scorpion and spider toxins.(AU)

Amazonian scorpions and scorpionism: integrating toxinological, clinical, and phylogenetic data to combat a human health crisis in the world's most diverse rainfores

Venom from Amazonian scorpions of the genus Tityus contains components capable of eliciting a distinct clinical, mostly neurological, syndrome. This contrasts with the mainly autonomic manifestations produced after envenomation by congeneric southern and northern South American species. Herein, we summarize Pan-Amazonian scorpionism by synthesizing available toxinological, clinical, and molecular data gathered from all affected areas in Amazonia, including Brazil, Ecuador, Colombia, Peru, Venezuela, and French Guiana. We searched multiple databases, as well as our own records, for reports of scorpion envenomations in Amazonia by confirmed Tityus spp., and compared the clinical manifestations. To help uncover clinical and venom relationships among problematic species, we explored phylogenetic relationships with a rate-calibrated analysis of mitochondrial COI data from available species. The possible existence of diversity gradients for venom toxic and immunogenic components despite the predicted strong phylogenetic association among species is underscored by discussed clinical and toxinological findings. A multicentric effort, involving all nations affected by this neglected disease, is urgently needed to offer alternatives for treating and understanding this pathology, including the preparation of neutralizing antibodies with a broad range of efficacy.(AU)

Scorpion stings in pregnancy: an analysis of outcomes in 66 envenomed pregnant patients in Iran

Scorpionism is one of the most important health problems in tropical regions, which unfortunately results in thousands of deaths annually. Pregnant women are potential victims in areas with high scorpion-sting prevalence. Limited medical data are available on the effects of scorpion envenomation in pregnant women. This study aimed to examine the effect of scorpion envenomation on pregnancy outcomes in 66 cases. Methods: The present descriptive/analytical cross-sectional study was performed on 66 scorpion-envenomed pregnant women referred to the clinical toxicology unit of Ahvaz Razi Hospital in Iran during 2015-2017. The variables assessed in all cases, via questionnaire and hospital medical records, were: age, patient residency, gestational week, status of the fetus, laboratory anomalies, clinical severity of envenomation, sting site and scorpion species. Pregnancy outcome (miscarriage, stillbirth, preterm birth, normal delivery) and status of the newborns were also evaluated. Data were analyzed using SPSS ® software (version 24.0). Results: The following pregnancy outcomes were recorded from envenomed pregnant women: miscarriage = 1.5% (n = 1), stillbirth = 4.5% (n = 3), preterm birth = 10.6% (n = 7), normal birth = 83% (n = 55). Among participants whose pregnancy led to birth, 11(17.7%) cases had prenatal-neonatal complications. Neonatal complications, including Apgar score less than 8 points at 5 min, were found in 7 (11.3%) preterm birth cases and in 4 (6.4%) normal birth cases, along with birth weight below 2500 g in normal births. A significant relationship was found between adverse pregnancy outcomes and bite location, as well as scorpion species, but no relationship was found with other variables. Conclusion: Envenomation significantly contributes to preterm birth. Moreover, the location of bites and the type of scorpion species have a decisive role in the pregnancy outcome of scorpion-envenomed pregnant women.(AU)

Achados necroscópicos em cães e gatos vítimas de intoxicação exógena / Necroscopic findings in dogs and cats victim of intentional intoxication

O reconhecimento adequado dos sinais clínicos e lesões anatomopatológicas são importantes para caracterizar os casos de intoxicação exógena em cães e gatos. O diagnóstico definitivo do agente tóxico deve ser realizado por exame toxicológico. No período de 2009 a 2014 foram analisados 42 casos de necropsia com histórico de intoxicação exógena de 31 (73,8%) cães e 11 (26,2%) gatos. Destes 21 (50%) apresentaram Boletim de Ocorrência e 22 (52,4%) foram submetidos ao exame toxicológico. A maior prevalência verificada foi intoxicação por carbamato e nos exames necroscópicos a principal causa de morte insuficiência cardiorrespiratória, seguida por choque hipovolêmico. Na histopatologia do fígado, rim e cérebro as principais lesões encontradas foram congestão, degeneração e hemorragia. Os órgãos apresentaram sinais de autólise e putrefação. Com base no estudo observou-se que as técnicas diagnósticas complementares, histopatologia e exame toxicológico, fornecem suporte adicional na elaboração de laudos em casos de intoxicações exógenas criminais envolvendo animais.(AU)

Appropriate recognition of the clinical signs, lesions anatomical pathological that characterizes cases of animals intoxication that take to death. Associated with the methods of identification laboratorial of forensic toxicology is fundamental to establish a definitive diagnosis of the toxic agent. In period 2009 to 2014 are selected 42 cases, 31 (73,8%) dogs and 11 (26,2%) cats. These cases 21 (50%) feature police reports and 22 (52,4%) are made toxicological exam. Highest prevalence was intoxication for carbamate. Necroscopic exam revealed that main cause of death were cardiac respiratory insufficiency and hypovolemic shock. In histopathology of liver, kidney and brain the main lesions are congestion, degeneration and bleeding. The organs showed signs of autolysis and putrefaction. The diagnostic techniques used, histopathology and necropsy are additional and helps veterinarian to make reports for litigation in cases of intentional poisoning in animals.(AU)

Laboratory animal welfare: environmental enrichment shows positive effect on animal testing / Bem-estar animal em laboratório: enriquecimento ambiental apresenta efeito positivo na experimentação animal

Reproductive and developmental toxicology has focused on the need to approach the effects of organism exposure to various drugs during pregnancy after the mid-50s, when the thalidomide tragedy stroke humanity. In recent decades, this area of study has developed a lot due to animal testing, raising awareness on the need to improve the quality of life of such animals. Therefore, this paper aims to investigate how the science of animal welfare can improve scientific research as a whole, including the reproductive and developmental toxicology fields, by emphasizing environmental enrichment in animal facilities. To do so, we conducted an integrative literature review on several quantitative and qualitative methodological approaches that are applicable to toxicology studies. Here, we present evidence that environmental enrichment improves animal welfare and prevents or reduces the negative effects of captive housing, which must be a principle of toxicological research for ethical, legal and scientific reasons.(AU)

Na toxicologia da reprodução e do desenvolvimento, a atenção necessária foi dada aos efeitos da exposição do organismo às inúmeras drogas durante o período gestacional somente após a metade dos anos 50, quando a tragédia da talidomida atingiu a humanidade. Assim, esta área alcançou desenvolvimento científico com a contribuição da experimentação animal nas últimas décadas. O uso de animais de laboratórios para a pesquisa científica expôs a necessidade de melhorar a qualidade de vida destas espécies. Portanto, este trabalho tem como objetivo investigar como a ciência do bem-estar animal pode melhorar a pesquisa científica como um todo, incluindo na área de toxicologia da reprodução e de desenvolvimento, enfatizando o enriquecimento ambiental em biotérios. Uma revisão integrativa de literatura foi realizada, incluindo abordagens quantitativas e qualitativas, quais podem ser aplicadas para estudos de toxicologia. Aqui, são mostradas evidências de que o enriquecimento ambiental melhora o bem-estar animal e previne ou reduz os efeitos negativos do cativeiro, qual deve um princípio da pesquisa toxicológica por razões éticas, argumentos legais e garantias científicas.(AU)

Laboratory animal welfare: environmental enrichment shows positive effect on animal testing / Bem-estar animal em laboratório: enriquecimento ambiental apresenta efeito positivo na experimentação animal

Reproductive and developmental toxicology has focused on the need to approach the effects of organism exposure to various drugs during pregnancy after the mid-50's, when the thalidomide tragedy stroke humanity. In recent decades, this area of study has developed a lot due to animal testing, raising awareness on the need to improve the quality of life of such animals. Therefore, this paper aims to investigate how the science of animal welfare can improve scientific research as a whole, including the reproductive and developmental toxicology fields, by emphasizing environmental enrichment in animal facilities. To do so, we conducted an integrative literature review on several quantitative and qualitative methodological approaches that are applicable to toxicology studies. Here, we present evidence that environmental enrichment improves animal welfare and prevents or reduces the negative effects of captive housing, which must be a principle of toxicological research for ethical, legal and scientific reasons.(AU)

Na toxicologia da reprodução e do desenvolvimento, a atenção necessária foi dada aos efeitos da exposição do organismo às inúmeras drogas durante o período gestacional somente após a metade dos anos 50, quando a tragédia da talidomida atingiu a humanidade. Assim, esta área alcançou desenvolvimento científico com a contribuição da experimentação animal nas últimas décadas. O uso de animais de laboratórios para a pesquisa científica expôs a necessidade de melhorar a qualidade de vida destas espécies. Portanto, este trabalho tem como objetivo investigar como a ciência do bem-estar animal pode melhorar a pesquisa científica como um todo, incluindo na área de toxicologia da reprodução e de desenvolvimento, enfatizando o enriquecimento ambiental em biotérios. Uma revisão integrativa de literatura foi realizada, incluindo abordagens quantitativas e qualitativas, quais podem ser aplicadas para estudos de toxicologia. Aqui, são mostradas evidências de que o enriquecimento ambiental melhora o bem-estar animal e previne ou reduz os efeitos negativos do cativeiro, qual deve um princípio da pesquisa toxicológica por razões éticas, argumentos legais e garantias científicas.(AU)

Intoxicações de cães por drogas recreativas: maconha e cocaína

A intoxicação de animais domésticos por drogas recreativas está presente na rotina de atendimento clínico dos médicos veterinários de pequenos animais. O cão é a espécie mais acometida, uma vez que é capaz de ingerir ou inalar acidentalmente drogas como a maconha e cocaína, responsáveis por uma sintomatologia neurológica e cardiovascular que requer rigoroso acompanhamento e tratamento veterinário para a obtenção de um prognóstico favorável. O objetivo do presente trabalho é realizar uma revisão de literatura com enfoque nas drogas maconha e cocaína, ressaltando as particularidades da espécie canina e possíveis tratamentos a serem instaurados. (AU)

Pet intoxication by recreational drugs is embedded in the routine of the small animal clinical practice. Dogs are the most common species to undergo such intoxications, due to its capability of ingesting or inhaling compounds such as marijuana and cocaine. These drugs are responsible for a cardiovascular and neurological syndrome, thus requiring strict follow-ups and treatments, to obtain a favorable prognosis. The aim of this study is to perform a bibliographical review emphasizing marijuana and cocaine intoxications on dogs, highlighting their particularities and possible treatments to be instituted. (AU)

Intoxicações de cães por drogas recreativas: maconha e cocaína

A intoxicação de animais domésticos por drogas recreativas está presente na rotina de atendimento clínico dos médicos veterinários de pequenos animais. O cão é a espécie mais acometida, uma vez que é capaz de ingerir ou inalar acidentalmente drogas como a maconha e cocaína, responsáveis por uma sintomatologia neurológica e cardiovascular que requer rigoroso acompanhamento e tratamento veterinário para a obtenção de um prognóstico favorável. O objetivo do presente trabalho é realizar uma revisão de literatura com enfoque nas drogas maconha e cocaína, ressaltando as particularidades da espécie canina e possíveis tratamentos a serem instaurados.

Pet intoxication by recreational drugs is embedded in the routine of the small animal clinical practice. Dogs are the most common species to undergo such intoxications, due to its capability of ingesting or inhaling compounds such as marijuana and cocaine. These drugs are responsible for a cardiovascular and neurological syndrome, thus requiring strict follow-ups and treatments, to obtain a favorable prognosis. The aim of this study is to perform a bibliographical review emphasizing marijuana and cocaine intoxications on dogs, highlighting their particularities and possible treatments to be instituted.

It is time for top-down venomics

The protein composition of animal venoms is usually determined by peptide-centric proteomics approaches (bottom-up proteomics). However, this technique cannot, in most cases, distinguish among toxin proteoforms, herein called toxiforms, because of the protein inference problem. Top-down proteomics (TDP) analyzes intact proteins without digestion and provides high quality data to identify and characterize toxiforms. Denaturing top-down proteomics is the most disseminated subarea of TDP, which performs qualitative and quantitative analyzes of proteoforms up to ~30 kDa in high-throughput and automated fashion. On the other hand, native top-down proteomics provides access to information on large proteins (> 50 kDA) and protein interactions preserving non-covalent bonds and physiological complex stoichiometry. The use of native and denaturing top-down venomics introduced novel and useful techniques to toxinology, allowing an unprecedented characterization of venom proteins and protein complexes at the toxiform level. The collected data contribute to a deep understanding of venom natural history, open new possibilities to study the toxin evolution, and help in the development of better biotherapeutics.(AU)

Highlights in the knowledge of brown spider toxins

Abstract Brown spiders are venomous arthropods that use their venom for predation and defense. In humans, bites of these animals provoke injuries including dermonecrosis with gravitational spread of lesions, hematological abnormalities and impaired renal function. The signs and symptoms observed following a brown spider bite are called loxoscelism. Brown spider venom is a complex mixture of toxins enriched in low molecular mass proteins (4-40 kDa). Characterization of the venom confirmed the presence of three highly expressed protein classes: phospholipases D, metalloproteases (astacins) and insecticidal peptides (knottins). Recently, toxins with low levels of expression have also been found in Loxosceles venom, such as serine proteases, protease inhibitors (serpins), hyaluronidases, allergen-like toxins and histamine-releasing factors. The toxin belonging to the phospholipase-D family (also known as the dermonecrotic toxin) is the most studied class of brown spider toxins. This class of toxins single-handedly can induce inflammatory response, dermonecrosis, hemolysis, thrombocytopenia and renal failure. The functional role of the hyaluronidase toxin as a spreading factor in loxoscelism has also been demonstrated. However, the biological characterization of other toxins remains unclear and the mechanism by which Loxosceles toxins exert their noxious effects is yet to be fully elucidated. The aim of this review is to provide an insight into brown spider venom toxins and toxicology, including a description of historical data already available in the literature. In this review article, the identification processes of novel Loxosceles toxins by molecular biology and proteomic approaches, their biological characterization and structural description based on x-ray crystallography and putative biotechnological uses are described along with the future perspectives in this field.(AU)

Meta-analysis of individual and combined effects of mycotoxins on growing pigs

Little is known about the toxicity of concomitantly occurring mycotoxins in pig diets. This study was conducted to evaluate, through meta-analysis, the individual and the combined effects of mycotoxins on pig performance. The meta-analysis followed three sequential analyses (graphical, correlation, and variance-covariance) based on a database composed of 85 published papers, 1,012 treatments and 13,196 animals. Contamination of diets with individual mycotoxins reduced (p<0.05) feed intake by 14 % and weight gain by 17 %, while combined mycotoxins reduced the same responses by 42 % and 45 %, respectively, in comparison with the non-challenged group. The correlation (p <0.05) between reduction in weight gain (G) and reduction in feed intake (FI) was 0.67 in individual challenges and 0.93 in combined challenges. The estimated G was 6 % in individual challenges and 7 % in combined challenges when FI was zero, suggesting an increase in the maintenance requirements of challenged animals. Most of G (58 % in individual challenges and 84 % in combined challenges) was attributed to the changes in feed efficiency. The association of mycotoxins enhances individual toxic effects and the FI is important in explaining the deleterious effects on the growth of challenged pigs.

Initial peptidomic profiling of Brazilian sea urchins: Arbacia lixula, Lytechinus variegatus and Echinometra lucunter

Background Sea urchins can be found throughout the Brazilian coast and are reported to be one of the major causes of marine accidents on the shoreline. Although not lethal, these accidents are reported to be extremely painful. In order to understand the toxinology of the Brazilian urchins, a peptidomic approach was performed aiming to characterize the naturally occurring peptides in both the coelomic fluid and the spine. Methods Animals were collected without gender distinction and samples of the coelomic fluid and spines extracted were analyzed by RP-HPLC and mass spectrometry for peptide de novo sequencing. Results Several peptides were identified either in the coelomic fluid or the spine extract (except for E. lucunter). The peptide sequences were aligned with public deposited sequences and possible functions were inferred. Moreover, some peptides can be cryptides, since their sequences were identified within functional proteins, for example thymosin from Strongylocentrotus purpuratus. Conclusions Although preliminary, the peptidomic approach presented here reports, for the first time, the abundance of novel biological molecules derived from these animals. The discovery of such molecules may be of potential biotechnological application, as described for other organisms; nevertheless, further studies are required.(AU)

Meta-analysis of individual and combined effects of mycotoxins on growing pigs

Little is known about the toxicity of concomitantly occurring mycotoxins in pig diets. This study was conducted to evaluate, through meta-analysis, the individual and the combined effects of mycotoxins on pig performance. The meta-analysis followed three sequential analyses (graphical, correlation, and variance-covariance) based on a database composed of 85 published papers, 1,012 treatments and 13,196 animals. Contamination of diets with individual mycotoxins reduced (p<0.05) feed intake by 14 % and weight gain by 17 %, while combined mycotoxins reduced the same responses by 42 % and 45 %, respectively, in comparison with the non-challenged group. The correlation (p <0.05) between reduction in weight gain (G) and reduction in feed intake (FI) was 0.67 in individual challenges and 0.93 in combined challenges. The estimated G was 6 % in individual challenges and 7 % in combined challenges when FI was zero, suggesting an increase in the maintenance requirements of challenged animals. Most of G (58 % in individual challenges and 84 % in combined challenges) was attributed to the changes in feed efficiency. The association of mycotoxins enhances individual toxic effects and the FI is important in explaining the deleterious effects on the growth of challenged pigs.(AU)

Effects of 4-Nonylphenol on reproduction of exposed females during puberty

4-Nonylphenol (NP) is considered an endocrine disruptor due to its capacity to interact with endocrine system, mimicking estrogen activity. It is aubiquitous pollutant since it has been found in many human tissue samples and its precursor identified in sewage, sediments, rivers and drinking water. Thus, this work aimed to evaluate reproductive parameters in female Balb/C mice treated with 50 mg/kg body weight of NP for 21 days, a period in which they reach sexual maturity. Body weight during treatment, day of vaginal opening, organs weight, fertility rate and proportion of different follicle types were some of these parameters. Additionally, for analysis of some effect in littermates, a group of animals underwent two attempts at pregnancy after treatment. NP had no effect on body weight during treatment, nor did it alter the day of vaginal opening and fertility rate of females. The proportion of preantral, antral and degenerated ovarian follicles was not modified by treatment with NP, and no morphological or pathological alterations were observed. The number of pups did not differ between treatments in any attempt at pregnancy. Also the weight of pups on birth and female rate in littermates was similar among experimental groups after both trials of pregnancy. It is suggested that the NP available or accumulated during the period of sexual maturation (21 days) did not reach threshold levels capable of producing significant alterations in the reproductive parameters analyzed.

Effects of 4-Nonylphenol on reproduction of exposed females during puberty

4-Nonylphenol (NP) is considered an endocrine disruptor due to its capacity to interact with endocrine system, mimicking estrogen activity. It is aubiquitous pollutant since it has been found in many human tissue samples and its precursor identified in sewage, sediments, rivers and drinking water. Thus, this work aimed to evaluate reproductive parameters in female Balb/C mice treated with 50 mg/kg body weight of NP for 21 days, a period in which they reach sexual maturity. Body weight during treatment, day of vaginal opening, organs weight, fertility rate and proportion of different follicle types were some of these parameters. Additionally, for analysis of some effect in littermates, a group of animals underwent two attempts at pregnancy after treatment. NP had no effect on body weight during treatment, nor did it alter the day of vaginal opening and fertility rate of females. The proportion of preantral, antral and degenerated ovarian follicles was not modified by treatment with NP, and no morphological or pathological alterations were observed. The number of pups did not differ between treatments in any attempt at pregnancy. Also the weight of pups on birth and female rate in littermates was similar among experimental groups after both trials of pregnancy. It is suggested that the NP available or accumulated during the period of sexual maturation (21 days) did not reach threshold levels capable of producing significant alterations in the reproductive parameters analyzed.(AU)

Concentrações de oleandrina nas folhas de Nerium oleander de diferentes cores da floração / Oleandrin concentrations in leaves of Nerium oleander of different colors of flowering

Nerium oleander é uma planta ornamental responsável por intoxicações em animais e humanos. Todas as partes da planta contém glicosídeos cardiotóxicos, principalmente a oleandrina. Alguns autores apontam que a toxicidade da planta apresentaria variação em função da cor da flor. O objetivo deste trabalho foi determinar se existe variação na concentração de oleandrina nas folhas de N. oleander de exemplares da planta com diferentes cores de inflorescências. Foram coletadas 10 amostras de folhas para cada tipo de flor (branca, rosea e vermelha). Os níveis de oleandrina foram determinados por meio de HPLC-UV após extração com metanol, precipitação com acetato de chumbo e separação em cartucho de C18. A média da concentração obtida em todas as folhas analisadas foi de 4,89mg g-1, sendo 6,20±4,08mg g-1 na variedade de flores brancas, 4,16±3,44mg g-1 na de flores roseas e 4,31±1,99mg g-1 na de flores vermelhas. As concentrações de oleandrina não apresentaram diferença estatisticamente significante entre as variedades de flores.(AU)

Nerium oleander is an ornamental plant responsible for poisoning in animals and humans. All parts of the plant contain cardiotoxic glycosides, mainly oleandrin. Some authors suggest that the toxicity of the plant would present a dependence on flower color. The objective of this study was to determine whether there is variation in the concentration of oleandrin in the leaves of N. oleander specimens of different colors of flowers. Ten samples of leaves from each variety of flower (white, pink and red) were collected. Oleandrin levels were determined by HPLC-UV after extraction with methanol, precipitation with lead acetate and separation with C18 cartridge. The average concentration obtained in all analyzed leaves was 4.89mg g-1, being 6.20±4.08mg g-1 in variety of white flowers, 4.16±3.44mg g-1 in pink flowers, and 4.31±1.99mg g-1 in red flowers. The concentrations of oleandrin did not show statistically significant difference between the varieties of flowers.(AU)

Thalassophryne nattereri fish venom: from the envenoming to the understanding of the immune system

Thalassophryne nattereri (niquim) is a venomous fish found off North and Northeast coast of Brazil, where it is known by the severity of the accidents involving humans. This review article is divided into four topics. The first one provides a brief description of the animal biology and its distribution off Brazilian coastal waters, the venom apparatus, signs and symptoms observed in envenomated humans and also describes envenomation in mice. The second topic describes the use of modern genetic approach and mass spectrometry for identification of highly expressed genes in its venom glands and the sequence of major toxins. The third chapter offers a detailed study of tissue injury induced by the venom and reveals the role of toxins that impair inflammation reduction. Finally, the fourth section expands the understanding of many extrinsic and intrinsic essential factors in maintaining survival of memory B cell compartment. Our results demonstrate the wide possibilities for research in the area of toxinology, also the necessity of interconnection among biochemistry, pharmacology and immunology areas for the expansion of knowledge and for generation of innovation.(AU)

Thalassophryne nattereri fish venom: from the envenoming to the understanding of the immune system

Thalassophryne nattereri (niquim) is a venomous fish found off North and Northeast coast of Brazil, where it is known by the severity of the accidents involving humans. This review article is divided into four topics. The first one provides a brief description of the animal biology and its distribution off Brazilian coastal waters, the venom apparatus, signs and symptoms observed in envenomated humans and also describes envenomation in mice. The second topic describes the use of modern genetic approach and mass spectrometry for identification of highly expressed genes in its venom glands and the sequence of major toxins. The third chapter offers a detailed study of tissue injury induced by the venom and reveals the role of toxins that impair inflammation reduction. Finally, the fourth section expands the understanding of many extrinsic and intrinsic essential factors in maintaining survival of memory B cell compartment. Our results demonstrate the wide possibilities for research in the area of toxinology, also the necessity of interconnection among biochemistry, pharmacology and immunology areas for the expansion of knowledge and for generation of innovation.

Suscetibilidade dos estágios ontogenéticos: padrão osmorregulatório em camarões peneídeos e palemonídeos / Susceptibility of ontogenetic stages: osmoregulatory standard in penaeid and palaemonid shrimps / Susceptibilidad de etapas ontogenéticas: estándar osmorregulatorio en camarones peneídeos y palemonídeos

A invasão da água doce por crustáceos ocorreu repetidamente ao longo dos anos, pressionando um ajuste em seus processos fisiológicos. A habilidade de sobreviver aos diferentes ambientes foi encontrada em traços osmorregulatórios, e estão relacionadas à capacidade de regular o fluido extracelular. Portanto, o objetivo deste manuscrito é explorar os processos osmorregulatórios de camarões palemonídeos e peneídeos, visualizando um padrão na capacidade osmorregulatória em ambas as famílias e, em particular, tratar de assuntos pertinentes aos efeito no ciclo ontogenético e o efeito ocasionado pelo nitrito na hemolinfa de crustáceos. Comparando palemonídeos e peneídeos, percebe-se que o sucesso da conquista do ambiente dulcícola, e a não necessidade de hiporregulação, levou à perda dessa capacidade na maioria dos palemonídeos. Dessa forma, se supõem um ancestral com capacidade hiporregulatória, uma vez que invertebrados marinhos são, parcimoniosamente, osmoconformadores. Compara-se também que o nitrito apresenta efeito nas proteínas presentes na hemolinfa dos crustáceos quando submetidos à exposição natural ou induzida deste composto, e que a capacidade osmorregulatória é, portanto, variada de acordo com seus estágios.(AU)

The invasion of freshwater by crustaceans has occurred repeatedly over the years, pressing for an adjustment in their physiological processes. The ability to survive in different environments was found in osmoregulatory traits and is related to the ability to regulate the extracellular fluid. Therefore, the purpose of this manuscript is to explore the osmoregulatory processes in palaemonid and penaeid shrimps, which display a pattern of osmoregulatory capacity in both families and, in particular, to deal with matters related to the effects on ontogenetic cycle and the effect raised by nitrite in crustacean hemolymph. By comparing palaemonid and peneid shrimps, it is possible to note the success of conquering freshwater environment, and not the need for hypo-regulation, has led to the loss of this capability in most palaemonids. Thus, it is assumed that they share an ancestor with hypo-regulatory capacity, since marine invertebrates are sparsely osmoconformers. The article also compares that nitrite has effect on proteins present in the hemolymph of crustaceans when subjected to natural or induced exposure to these compound, and osmoregulatory capacity is therefore varied according to their stages.(AU)

La invasión del agua dulce por crustáceos ocurre repetidamente a lo largo de los años, pulsando un ajuste en sus procesos fisiológicos. La capacidad de sobrevivir en diferentes ambientes se ha encontrado en rasgos osmorregulatorios y están relacionados a la capacidad de regular el fluido extracelular. Por lo tanto, el objetivo de este manuscrito es el de explotar los procesos osmorregulatorios de camarones palemonídeos y peneídeos, se muestra un estándar en la capacidad osmorregulatoria en ambas las familias y, en especial, trata de cuestiones relativas a los efectos sobre los ciclo ontogenético y el efecto causado por nitrito en la hemolinfa de crustáceos. Comparando palemonídeos y peneídeos, se da cuenta de que el éxito de la conquista del ambiente dulcícola, y sin necesidad de hipo regulación, condujo a la pérdida de esa capacidad en la mayoría de los palemonídeos. Así, se supone un ancestral con capacidad hipo regulatoria, una vez que los invertebrados marinos son, parsimoniosamente, osmoconformadores. Se compara también que el nitrito presenta efectos en las proteínas presentes en la hemolinfa de los crustáceos cuando sometidos a la exposición natural o inducida por estes compuesto, y que la capacidad osmorregulatoria es, por lo tanto, variada de acuerdo con sus etapas.(AU)

Acute copper toxicity in juvenile fat snook Centropomus parallelus (Teleostei: Centropomidae) in sea water

Three experiments were designed to assess the accumulation and acute toxicity of copper (Cu) in juvenile fat snook Centropomus parallelus. The first experiment was performed to determine the 96-h lethal concentration (LC50) of Cu. The second experiment was designed to assess the effects of sublethal concentrations of Cu (0.47 and 0.94 mg/L), while the third one allowed us to test the recovery capacity of fish exposed to the sublethal concentrations Cu and kept in sea water without Cu addition. The LC50 value for Cu was found to be 1.88 mg/L Cu. Fish exposed to the sublethal concentrations of Cu showed a significant accumulation of Cu in gills at 96 h respect to the control ones (0.43 µg/g Cu). No significant difference was observed in the accumulation of Cu in gills between fish exposed to 0.47 mg/L (1.09 µg/g Cu) and 0.94 mg/L (1.26 µg/g Cu). Exposure (24 and 96 h) to the sublethal concentrations of Cu tested induced DNA damage in the erythrocytes. The results show that acute exposure to sublethal concentrations induces Cu accumulation and DNA damage in fish, these effects being recovered after 240 h in sea water without Cu addition.(AU)

Três experimentos foram realizados para avaliar o acúmulo e toxicidade aguda do cobre (Cu) em juvenis de robalo-peva Centropomus parallelus. O primeiro experimento foi realizado para determinar a concentração letal (96h-CL50) de Cu. O segundo experimento foi realizado para avaliar os efeitos de concentrações subletais de Cu (0,47 e 0,94 mg/L), enquanto o terceiro permitiu testar a capacidade de recuperação dos peixes expostos a concentrações subletais do Cu e posteriormente mantidos em água do mar sem acréscimo de Cu. O valor de LC50 encontrado para o Cu foi de 1,88 mg/L. Os peixes expostos as concentrações subletais de Cu mostraram um acúmulo significativo nas brânquias em relação ao controle em 96 h de exposição (0,43 µg/g Cu). Nenhuma diferença significativa foi observada entre os peixes expostos a 0,47 mg/L de Cu (1,09 µg/g) e 0,94 mg/L de Cu (1,26 µg/g). A exposição (24 e 96 h) para as concentrações subletais de Cu induziram danos no DNA. Os resultados mostram que a exposição aguda a concentrações subletais induz o acúmulo de Cu e danos ao DNA nas brânquias dos peixes, onde estes efeitos são recuperados após 240 h em água do mar sem adição de Cu.(AU)