Multi-biomarker Assessment in a Native Species Psalidodon eigenmanniorum Under Inorganic Mercury and Recovery Scenarios.

The increasing contamination of water bodies with mercury raises concerns about its possible effects on aquatic organisms. The combined use of several biomarkers allows researchers to study the impact of a chemical at different levels of biological organization. In the present work, we determined the response of histological (gills and liver), somatic (condition factor and hepato-somatic index), and behavioral (predator-prey relationship, through the presentation of a computer-animated image) biomarkers in the native species Psalidodon eigenmanniorum exposed to 100 µg L-1 of inorganic Hg (IHg) during 96 h. We also assessed whether there was a change in the biomarkers analyzed after 7 days in Hg-free water compared with those exposed to IHg. In exposed fish, IHg caused damage to the gills and liver tissues. The condition factor showed no difference between IHg-exposed organisms and control organisms, while the hepato-somatic index was lower in IHg-exposed fish. As for the behavioral analyses, it was observed that the presentation of a stimulus induced changes in the behavioral responses of fish exposed to IHg, which showed a heightened state of alertness with respect to control. On the other hand, after 7 days in Hg-free water, the organisms generally showed no changes in biomarkers compared with IHg-exposed fish. Our results contribute new data on IHg toxicity in a native species and provide information on the plasticity of damage to reverse itself. Furthermore, this work provides baseline information for environmental assessments in water bodies where mercury is present.

What types of enzyme activities are useful biomarkers of bifenthrin exposure on Chironomus sp. (Diptera, Chironomidae) larvae under laboratory and field-based microcosm conditions?

Bifenthrin is a second generation synthetic pyrethroid insecticide that is widely used in Australia and worldwide. It is frequently found in urban freshwater sediments at concentrations likely to impact biota as it is highly toxic to fish and macroinvertebrates, such as chironomids. Our main goal was to evaluate if oxidative stress and hydrolase enzymes are useful biomarkers of effect of synthetic pyrethroids exposure under different scenarios. Chironomus tepperi larvae (5 days old) were exposed to sub-lethal sediment concentrations of bifenthrin for 5 days under controlled laboratory conditions. A field-based microcosm exposure with bifenthrin-spiked sediments (using the same concentrations as the laboratory exposure) was carried out at a clean field site for four weeks to allow for colonization and development of resident chironomid larvae. At the end of both experiments, Chironomus larvae (C. tepperi in the laboratory exposures and C. oppositus in the microcosm exposures) were collected and oxidative stress enzymes (Glutathione-s-Transferase, Glutathione Reductase and Glutathione Peroxidase) and hydrolase enzymes (Acetylcholinesterase and Carboxylesterase) were measured. Only the Glutathione Peroxidase activity was significantly impacted in larvae from the laboratory exposure. On the contrary, significant changes were observed in all the measured enzymes from the field-based microcosm exposure. This is likely because exposure was throughout the whole life cycle, from egg mass to fourth instar, showing a more realistic exposure scenario. Furthermore, this is the first time that changes in oxidative stress and hydrolase enzymes have been shown to occur in Australian non-biting midges exposed under field-based microcosm conditions. Thus, this study demonstrated the usefulness of these enzymes as biomarkers of effect following bifenthrin exposure in microcosms. It also highlights the importance of using a range of different biochemical endpoints to get a more holistic understanding of pesticide effects and the pathways involved.

Spermatozoa characterization in the one-sided livebearing Jenynsia multidentata (Cyprinodontiformes: Anablepidae).

Several sperm parameters have been employed as useful tools to evaluate fish fertility. Within teleosts, approximately 3% of fish species are known to be viviparous. The Order Cyprinodontiformes includes several species with internal fertilization, and within this group most of the studies about sperm quality have been mainly focused on the Poeciliidae family. The livebearing fish Jenynsia multidentata (Anablepidae) inhabits an extensive area of the Neotropical region and it has been used as a useful fish laboratory model to evaluate the effects of xenobiotics through different biomarkers. The present work characterized the sperm of this species through a simple protocol of semen collection. Sperm population showed linearity greater than 89% and 70% of fish have a straight line and curvilinear velocity valued between 50 and 100 µm/s. Although 85% of individuals showed a proportion of live sperm higher than 60%, the male population had a high degree of heterogeneity in its sperm count. Morphometry analyses showed a total sperm and head lengths of 46.66 ± 2.06 µm and 3.46 ± 0.41 mm, respectively. A rather long midpiece region (9.12 ± 0.65 µm) was registered, which may indicate high energy-producing capabilities of the spermatozoa. This study established basic parameter values which could be useful for evaluating reproductive potential of J. multidentata populations.

Spermatozoa characterization in the one-sided livebearing Jenynsia multidentata (Cyprinodontiformes: Anablepidae) / Caracterización de parámetros espermáticos del pez vivíparo Jenynsia multidentata (Cyprinodontiformes: Anablepidae)

Several sperm parameters have been employed as useful tools to evaluate fish fertility. Within teleosts, approximately 3% of fish species are known to be viviparous. The Order Cyprinodontiformes includes several species with internal fertilization, and within this group most of the studies about sperm quality have been mainly focused on the Poeciliidae family. The livebearing fish Jenynsia multidentata (Anablepidae) inhabits an extensive area of the Neotropical region and it has been used as a useful fish laboratory model to evaluate the effects of xenobiotics through different biomarkers. The present work characterized the sperm of this species through a simple protocol of semen collection. Sperm population showed linearity greater than 89% and 70% of fish have a straight line and curvilinear velocity valued between 50 and 100µm/s. Although 85% of individuals showed a proportion of live sperm higher than 60%, the male population had a high degree of heterogeneity in its sperm count. Morphometry analyses showed a total sperm and head lengths of 46.66±2.06µm and 3.46±0.41mm, respectively. A rather long midpiece region (9.12±0.65µm) was registered, which may indicate high energy-producing capabilities of the spermatozoa. This study established basic parameter values which could be useful for evaluating reproductive potential of J. multidentata populations.

Diversos parámetros espermáticos han sido utilizados para evaluar la fertilidad de peces. Dentro de los peces teleósteos, aproximadamente el 3% de las especies son vivíparas. El orden Cyprinodontiformes incluye varias especies con fecundación interna. Dentro de este orden la mayor parte de los estudios sobre la calidad del esperma se han centrado principalmente en la familia Poeciliidae. El pez vivíparo Jenynsia multidentata (Anablepidae) habita una extensa área de la región Neotropical y ha sido utilizado como un exitoso modelo de laboratorio. El objetivo del presente trabajo fue caracterizar los espermatozoides de esta especie a través de un simple protocolo de recolección de esperma. La población de espermatozoides mostró una linealidad superior al 89% y el 70% de los peces tienen una velocidad lineal y curvilineal entre 50 y 100µm/s. Aunque el 85% de los individuos mostró una proporción de espermatozoides vivos de más del 60%, se observó una alta heterogeneidad en el recuento espermático. Los análisis morfométricos mostraron una longitud total de espermatozoides de 46.66±2.06µm y una longitud de la cabeza de 3.46±0.41µm. Los espermatozoides presentan una pieza media larga (9.12±0.65µm) lo que puede indicar una alta capacidad de producción de energía. El presente estudio establece valores básicos de parámetros que pueden ser útiles para evaluar el potencial reproductivo de las poblaciones de J. multidentata.

Impairments in aromatase expression, reproductive behavior, and sperm quality of male fish exposed to 17ß-estradiol.

Growing evidence shows that environmental estrogen can reach levels that are high enough to exert adverse reproductive effects on wild fish populations. The authors report different parameters of male reproductive behavior, brain, and gonadal aromatase expression, as well as sperm quality in an internally fertilizing fish species (Jenynsia multidentata, Jenyns) exposed to environmentally relevant concentrations of 17ß-estradiol (E(2) ). Adult males were exposed to 0, 50, 100, and 250 ng/L E(2) over 28 d. The authors' findings demonstrate that E(2) exposure resulted in a very clear increase in brain aromatase transcript abundance at all assayed concentrations compared with control; however, no effects on gonadal aromatase expression were observed. Behavioral measures revealed increased sexual activity at 50 ng/L but not 100 or 250 ng/L E(2) . In contrast to the molecular and behavioral responses, the condition factor, gonadosomatic index, and sperm quality were unaltered by E(2) exposure. The results from the present work suggest that E(2) affects some aspects of the reproductive biology of J. multidentata. These modifications in the reproductive biology caused by exposure to E(2) could potentially lead to long-term effects at population levels that may not always be immediately evident. To the best of the authors' knowledge, this is the first report on the combined effect of E(2) on aromatase expression, sexual behavior, and sperm parameters in fish.

Changes in the swimming activity and the glutathione S-transferase activity of Jenynsia multidentata fed with microcystin-RR.

We report the effects of sublethal doses of microcystin-RR (MC-RR) on the swimming activity of Jenynsia multidentata as well as the simultaneous response of its detoxication system by measuring glutathion S-transferase (GST) activities in the liver and brain of fish. MC-RR was applied on the food pellets at doses of 0.01, 0.1 and 1 microg g(-1). Swimming activity was recorded 10 min each hour over 24h by using a computer-based image processing system, which facilitates quantification of two measures of fish swimming behaviour (average velocity, movement percentage). Results show that low levels of cyanotoxin increased the swimming activity, while the highest dose used produced significant changes with respect to control group only since approximately 20 h of exposure, when the swimming activity was decreased. On the other hand, GST activity was significantly increased only in the liver and brain of fish fed with the highest MC-RR dose. Both results suggest that fish are reacting to the stress caused by low doses of MC-RR by increasing their swimming activity, raising further questions on the probable neurotoxicity of MCs, and presenting the behavioural change as a good biomarker of early toxic stress. On the other hand, fish reduced their swimming speed at the highest MC-RR dose, when the detoxication activity began, which can be hypothesized to be a reallocation of their energy, favouring detoxication over swimming activity.

Endosulfan acute toxicity and histomorphological alterations in Jenynsia multidentata (Anablepidae, Cyprinodontiformes).

Toxicity tests using adult specimens of Jenynsia multidentata were carried out during 96 hours in order to determine the lethal concentration (LC50) of endosulfan. Histological alterations were determined in gills and liver. Gill damage was quantified as secondary lamellae thickness. The 96 hr LC50 values were significantly different between males (0.719 microg x L(-1)) and females (1.317 microg x L(-1)). The sex difference was attributed to the dimorphism in the lipid content in females (2.16%) and males (1.79%). Histological alterations in gills included hypertrophy and lifting of the epithelium of the secondary lamellae and aneurisms. These alterations caused a significant increase of the secondary lamellae thickness in treatment versus control fish. Finally, reversible histological alterations (such as hydropic degeneration and dilation of sinusoids) were observed in the liver of exposed fish as well as an irreversible change such as necrosis at the highest concentrations.