Cryopreservation: Extending the viability of biological material from sea urchin (Echinometra lucunter) in ecotoxicity tests.

The sea urchin, Echinometra lucunter, is widely used in embryo-larval tests for ecotoxicological studies in Brazil and other countries. For each test, sea urchins are collected from the wild and this can cause impact on wild populations and it is limited by the weather and season which in turn limits the ability to carry out the tests. Cryopreservation is a method of live biological material storage at low temperature and can be used for long periods with little decline in viability, reducing the number of animals taken from the wild and enabling testing to be carried out on demand, irrespective of spawning season or location. In this study, 15 combinations of cryoprotective agents (CPAs) were evaluated on spermatozoa, subjected to a rapid cooling curve followed by immersion in liquid nitrogen. Twenty-four CPA combinations were evaluated on eggs subjected to a more gradual cooling curve in nitrogen vapor down to -35 °C and then plunging in liquid nitrogen. Fertilization tests using cryopreserved spermatozoa gave high pluteus larvae yields (≈80%) when concentrations of 10.5% or 13.65% ME2SO or 13.65% ME2SO+15.75% sucrose were used. The higher concentrations of ME2SO plus sucrose were more effective at maintaining the fertilization capacity of spermatozoa post-thawing. Egg cryopreservation was not successful with 0% fertilization observed post-thawing. The results suggest that it is feasible to implement spermatozoa cryopreservation as technological innovation to create a sperm bank for E. lucunter, which can be used in ecotoxicological tests, bringing benefits for researches and contributing to the conservation of the species.

Temperature affects the hypoxia tolerance of neotropical Cichlid Geophagus brasiliensis

Oxygen and temperature are the most limiting factors in aquatic environments. Several species are exposed to variations of these factors in water because of physical, chemical and biological processes. The objective of this study was to evaluate the metabolic profile and the tolerance to the hypoxia of Geophagus brasiliensis exposed to changes in temperature and oxygen availability. The fish were exposed to 20 and 90% of oxygen saturation combined with different temperatures (20°, 24° and 28° C) for 8 h. Hepatic and muscular glycogen, as well as the activities of lactate dehydrogenase (LDH), malate dehydrogenase (MDH), citrate synthase (CS) and their ratios were evaluated. Both glycogen and MDH activity showed a significant difference in the liver. While CS showed increased activity only in the heart. The increase in LDH activity in the white muscle shows the importance of the anaerobic pathway as energy source in this tissue. The MDH / LDH ratio increased in all tissues, while CS / LDH increased in the liver and decreased in the heart. Based on the results of the present study it may be concluded that this species used the anaerobic metabolism as the main strategy for hypoxia tolerance.(AU)

O oxigênio e a temperatura são os fatores mais limitantes em ambientes aquáticos. Várias espécies são expostas a variações destes fatores na água como resultado de processos físicos, químicos e biológicos. O estudo objetivou avaliar o perfil metabólico e a tolerância à hipóxia de Geophagus brasiliensis expostos a alterações na temperatura e disponibilidade de oxigênio. Os peixes foram expostos a 20% e 90% de saturação de oxigênio combinadas com diferentes temperaturas (20 ° C, 24 ° C e 28 ° C) durante 8h. Foram avaliados o glicogénio hepático e muscular, assim como as atividades das enzimas lactato desidrogenase (LDH), malato desidrogenase (MDH), citrato sintase (CS) e suas razões. Tanto o glicogênio quanto a atividade da MDH apresentaram diferença significativa no fígado. Enquanto a CS apresentou aumento de sua atividade apenas no coração. O aumento da atividade LDH no músculo branco mostra a importância da via anaeróbia como fonte de energia neste tecido. A razão MDH/LDH aumentou em todos os tecidos, enquanto CS/LDH apresentou aumento no fígado e diminuição no coração. Os resultados obtidos permitem concluir que esta espécie utilizou o metabolismo anaeróbio como principal estratégia de tolerância à hipóxia.(AU)

Anesthetic induction and recovery time of Centropomus parallelus exposed to the essential oil of Aloysia triphylla / Indução e recuperação anestésica de Centropomus parallelus expostos ao óleo essencial de Aloysia triphylla

ABSTRACT: This study analyzed the effectiveness of the essential oil of Aloysia triphylla (EOA) as an anesthetic and stress reducing agent in the transport of fat snook ( Centropomus parallelus ). Juveniles were exposed to different concentrations of EOA (seawater-adapted: 25-300μL L-1; freshwater-adapted: 200μL L-1) to identify the anesthetic induction and recovery times. In an additional experiment, seawater-adapted fat snook were transported in plastic bags with 10 or 20μL L-1 EOA for 6 or 24h. The increased concentration of EOA proportionally decreased the time required for anesthesia induction. Mortality was 30 and 70% in fat snook transported with 10 and 20µL L-1, respectively, 24h after transport. The addition of 20µL L-1 EOA significantly reduced plasma cortisol levels during transport compared to the control group, but increased blood glucose levels after 6 and 12h of transport. The EOA reduced total ammonia and dissolved oxygen levels at the end of transport compared to control fish. The EOA is recommended for sedation (25-50µL L-1) and anesthesia (100-300µL L-1) but not for transport of fat snook because it increased mortality.

RESUMO: Este estudo verificou a eficácia do óleo essencial de Aloysia triphylla (OEA) como um anestésico e redutor de estresse no transporte de robalo-pevas ( Centropomus parallelus ). Os juvenis foram expostos a diferentes concentrações de OEA (adaptados à água do mar: 25-300µL L-1; adaptados à água doce: 200µL L-1) para identificar os tempos de indução e recuperação da anestesia. Em um experimento adicional, robalos-peva adaptados à água do mar foram transportados em sacos plásticos com 10 ou 20µL L-1 OE por 6 ou 24h. O aumento da concentração do OEA diminuiu proporcionalmente o tempo necessário para a indução da anestesia. A mortalidade foi de 30 e 70% em robalo-pevas transportados com 10 e 20µL L-1, respectivamente, 24h após o transporte. A adição de 20µL L-1 OEA reduziu significativamente o cortisol plasmático durante o transporte, comparado ao grupo controle, mas aumentou os níveis de glicose sanguínea após 6 e 12h de transporte. O OEA reduziu os níveis de amônia total e de oxigênio dissolvido ao final do transporte em comparação com o grupo controle. O OEA é recomendado para sedação (25-50µL L-1) e anestesia (100-300µL L-1), mas não para o transporte de robalo-peva, porque aumentou a mortalidade.

Genetic and biochemical effects induced by iron ore, Fe and Mn exposure in tadpoles of the bullfrog Lithobates catesbeianus.

For decades, the extraction of minerals has intensified in order to meet the demand of industry. Iron ore deposits are important sources of metals, such as iron (Fe) and manganese (Mn). The particulate ores can be dispersed during extraction, transport and storage, with potential to induce biological impacts. Amphibians are very sensitive to environmental stressors. Therefore, the present study aimed to assess the effects of iron ore, Fe and Mn exposure during the metamorphosis of Lithobates catesbeianus. Endpoints analyzed included morphological (biometrical and developmental analyses), whole body Fe and Mn concentration in, plasma ferritin concentration, erythrocyte DNA damage (measured through comet assay and micronucleus test) and liver activity of enzymes involved in oxidative status [glutathione S-transferase (GST) and catalase (CAT)]. Tadpoles were kept under control condition (no contaminant addition) or exposed to iron ore (3.79mg/L as fine particulate matter); Fe (nominal concentration: 0.51mg/L Fe as C10H12FeN2NaO8; Fe-EDTA); and Mn (nominal concentration: 5.23mg/L Mn as 4H2O.MnCl2) for 30 days. Virtually, no mortality was observed, except for one tadpole found dead in the iron ore treatment. However, tadpoles exposed to iron ore had longer tail than those kept under control conditions while tadpoles exposed to manganese chloride showed higher body length than control ones. Exposure to Fe and Mn induced a delay in tadpole metamorphosis, especially when these metals are presented not as a mixture (iron ore). Tadpoles exposed to iron ore had increased whole body Fe and Mn while those exposed to Fe and Mn accumulated each metal individually. Tadpoles exposed to any of the contaminants tested showed a significant increase in erythrocyte DNA damage and frequency of micronuclei. In addition, they showed higher liver GST activity respect with those kept under control conditions. Plasma ferritin concentration and liver CAT activity were higher only in tadpoles exposed to iron ore. These findings indicated that tadpoles accumulated Fe and Mn at the whole body level after exposure to the single metals or to their mixture as iron ore. In addition, they indicate that Fe and Mn accumulation can induce oxidative stress with consequent significant developmental, genotoxic and biochemical effects in L. catesbeianus tadpoles.