Fish Jenynsia Multidentata Research Articles

Glyphosate is Harmful to Early Life Stages of the Viviparous Fish Jenynsia Multidentata: Biochemical and Locomotor Effects.

Glyphosate is the most widely used herbicide worldwide due to its efficacy in weed control in agriculture. This herbicide has been consistently detected in the aquatic environment, causing harmful consequences to nontarget organisms residing in agricultural regions. In this study, we assessed the effects of environmentally relevant concentrations of glyphosate (30-100µg/L) on the early life stages of the viviparous fish Jenynsia multidentata through biochemical and locomotor endpoints. At 96h of exposure, 30 and 65µg/L glyphosate caused an increase in acetylcholinesterase (AChE) activity, and 65µg/L glyphosate also augmented the levels of lipid peroxidation. Glyphosate at 100µg/L did not alter the activity of acetylcholinesterase or the levels of lipid peroxidation, but it stimulated the activity of the cellular detoxification enzyme glutathione S-transferase. In addition, all concentrations affected the swimming of the fish. Under light conditions, glyphosate caused hypolocomotion at all concentrations tested, whereas under dark conditions, this was observed at 30 and 100µg/L. Hyperlocomotion was observed at 65µg/L glyphosate. These findings are alarming for the health of fish, such as J. multidentata that inhabit streams that pass through agricultural areas, especially for the early life stages of these fish. Research studying the effects of pollutants on native species is relevant to improve regulation that protects aquatic ecosystems.

Tissue‐specific bioconcentration and biotransformation of cypermethrin and chlorpyrifos in a native fish (Jenynsia multidentata) exposed to these insecticides singly and in mixtures

The aim of the present study was to evaluate the accumulation of cypermethrin and chlorpyrifos when the fish Jenynsia multidentata was exposed to these pesticides singly and in technical and commercial mixtures. Adult female fish were exposed over 96 h to 0.04 μg/L of cypermethrin; 0.4 μg/L of chlorpyrifos; 0.04 μg/L of cypermethrin + 0.4 μg/L of chlorpyrifos in a technical mixture; and 0.04 μg/L of cypermethrin + 0.4 μg/L of chlorpyrifos in a mixture of commercial products. Fish exposed to cypermethrin accumulated this compound only in muscle, probably because of the low biotransformation capacity of this organ and the induction of cytochrome P4501A1 (CYP1A1) expression in the liver. The accumulation of chlorpyrifos occurred in fish exposed to the insecticide (intestine > liver > gills) even when these fish had higher gluthatione-S-transferase (GST) activity in gills and P-glycoprotein (P-gp) expression in the liver, compared with the control. Fish exposed to the technical mixture showed cypermethrin accumulation (liver > intestine > gills) with higher levels than those measured in fish after only cypermethrin exposure. Higher expression levels of CYP1A1 in the liver were also observed compared with the Control. Fish exposed to the commercial mixture accumulated both insecticides (cypermethrin: intestine > gills and chlorpyrifos: liver > intestine > gills > muscle). In the organs where accumulation occurred, biotransformation enzymes were inhibited. Consequently, the commercial formulation exposure provoked the highest accumulation of cypermethrin and chlorpyrifos in J. multidentata, possibly associated with the biotransformation system inhibition. Environ Toxicol Chem 2017;36:1764-1774. © 2016 SETAC.

Effects of carbamazepine on cortisol levels and behavioral responses to stress in the fish Jenynsia multidentata

Carbamazepine (CBZ) is an anticonvulsant drug, prescribed worldwide for the treatment of epilepsy, bipolar disorder and trigeminal neuralgia, which has been frequently detected in aquatic environments. The objective of this study was to analyze if CBZ modifies scototaxis and shoaling behaviors and/or whole-body cortisol levels of the one-sided livebearing fish Jenynsia multidentata under stress condition. Female adults of J. multidentata were exposed to 0, 10, 50 and 200μgCBZ/L during 14days. After CBZ exposure, fish were subjected to restraint stress during 15min. Control animals were not exposed to CBZ or stress. In the light/dark preference test (scototaxis), the individuals under acute restraint stress (without CBZ) exhibited a significant increase in the mean speed and in the time spent both in the light compartment and in the bottom of the tank with respect to controls. They also showed a tendency to stay longer frozen in the light compartment. Fish exposed to 10 and 50μgCBZ/L showed a significant reduction in mean speed compared to stressed fish without CBZ. A reduction in the time spent in the bottom of the tank was also observed in fish exposed to 10μgCBZ/L. Fish exposed to 200μgCBZ/L showed a decreasing tendency in all behavioral endpoints (time spent in the light compartment, mean speed, time spent at the bottom and freezing) in comparison to stressed fish not exposed to CBZ. Considering whole-body cortisol results, fish under acute restraint stress (without CBZ) significantly increased their hormone levels with respect to the control group, while fish exposed to CBZ and acute restraint stress, significantly decreased their whole-body cortisol levels. There were no significant changes in shoaling behavior due to either stress or CBZ exposure and no significant differences in whole-body cortisol levels between experimental groups. Considering that the light/dark and shoaling tests measure different stress response behaviors regulated by different neuroendocrine systems, these results could indicate that CBZ has a differential effect on fish behavioral stress response and cortisol levels, depending on the behavioral test used and stressor applied.

The effects of osmolality on sperm quality in Jenynsia multidentata (Cyprinodontiformes: Anablepidae).

Sperm quality tests on fish are classically used for evaluating cryopreservation procedures, and they are also promising to assess aquatic toxicity and biomarkers of xenobiotic effects on reproduction. Osmotic shock from the storage medium is one of the main factors affecting sperm quality during evaluation. Thus, the objective of this study was to evaluate the effects of different osmolalities (240-460 mOsm/kg) for at least 4 days on the sperm quality parameters of the viviparous fish Jenynsia multidentata. The level of significance was (P < 0.05). The plasma osmolality of J. multidentata is 326 ± 3.9 mOsm/kg. The motility of fresh semen was higher in osmolalities of 280 and 300 mOsm/kg but did not differ between osmolalities from 240 to 320 mOsm/kg. Above 380 mOsm/kg, the motility observed was 0%. Over the time period studied motility increased with increasing osmolality, and the most constant and long-lasting rates were between 300 and 320 mOsm/kg. On the 4th day of evaluation, higher membrane integrity rates were observed between 280 and 360 mOsm/kg, higher mitochondrial membrane potential was observed between 300 and 460 mOsm/kg, and higher DNA integrity rates were observed between 260 and 380 mOsm/kg. Moreover, osmolalities ≥460 and ≤240 resulted in the lowest motility and DNA integrity levels. Over 4 days, the plasma membrane integrity was significantly lower at ≤260 and ≥400 mOsm/kg, and the mitochondrial membrane potential was significantly lower only in osmolalities ≤240 mOsm/kg. Therefore, we conclude that for sperm quality preservation in J. multidentata, an osmolality of 300-320 mOsm/kg of the most suitable diluent is necessary. Furthermore, we conclude that the storage of sperm in a hyposmotic (<260 mOsm/kg) or hyperosmotic (>400 mOsm/kg) solution affects not only motility but also other sperm quality parameters.

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 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.

Antioxidant, phase II and III responses induced by lipoic acid in the fish Jenynsia multidentata (Anablapidae) and its influence on endolsulfan accumulation and toxicity

Antioxidants like lipoic acid (LA) are known to trigger augmented antioxidant and phase II and III responses. This study aimed to evaluate the effect of LA in P-glycoprotein (Pgp) expression, glutathione-S-transferase (GST) activity, total antioxidant competence, levels of lipid peroxides (TBARS) and accumulation of the organochlorine insecticide endosulfan (Endo: α-, β-isomers and sulfate metabolite) in different organs of the fish Jenynsia multidentata. One hundred and twenty females (1.55±0.07g) were fed during 8days with (n=60) or without (n=60) a LA enriched ration (6000mg/kg). Four experimental groups were defined: −LA/−Endo; +LA/−Endo; −LA/+Endo; and +LA/+Endo. Endo groups were exposed during 24h to 1.4μg of insecticide/L. Results showed that only LA induced a significant increment in liver Pgp expression. GST activity was augmented in liver after exposure to LA or Endo. TBARS levels were lowered in liver and gills after LA pre-treatment. Total antioxidant capacity was lowered in liver of Endo exposed fish, a result that was reversed by LA pre-treatment. It is concluded that LA induced the expected effects in terms of Pgp expression, GST activity and reduced TBARS levels although favored α-Endo accumulation in brain. However, the Endo metabolism to the more persistent endosulfan sulfate was not facilitated by LA pre-treatment.

A method to measure total antioxidant capacity against peroxyl radicals in aquatic organisms: Application to evaluate microcystins toxicity

Determination of total antioxidant capacity, instead of the measurements of limited number of antioxidants, is very important for the understanding of how antioxidants interact with reactive oxygen species (ROS). Several techniques already exist with this propose, although some of them are extremely time-consuming. A new methodology is proposed, based on the detection of ROS by fluorometry (ex/em: 485/520 nm) employing 2′,7′-dichlorofluorescein diacetate (H 2DCF-DA) as substrate. Supernatant of homogenized samples from different organs (gill, muscle, liver, and brain) of the teleost fish Jenynsia multidentata (Anaplebidae) were exposed to peroxyl radicals generated by thermal (35 °C) decomposition of 2,2′-azobis (2 methylpropionamidine) dihydrochloride (ABAP, 4 mM). Different protein concentrations (0.5, 1, 2 and 8 mg/ml) were assayed to get the best signal and curve fitting of fluorescence data over time (30 min). Total antioxidant capacity against peroxyl radicals was estimated as the difference in ROS area with and without ABAP, relative to the fluorescence registered without ABAP. For application of this methodology, J. multidentata specimens were exposed for 24 h to microcystins, cyanotoxins known to induce oxidative stress. Almost all organs showed a lower antioxidant capacity ( p < 0.05) in samples with 8 mg proteins/ml, when compared to protein content of 1–2 mg/ml. In liver samples, higher ( p < 0.05) free iron content was determined in samples with 8 mg proteins/ml. Sensitivity test employing GSH spiked in homogenized samples showed the protocol efficiency in detecting total antioxidant capacity. In the test with microcystins a dose-dependent decrease ( p < 0.05) of antioxidant competence in gills and brain and an inverse result with liver samples were observed. The use of antioxidant defenses was efficient in avoiding oxidative damage, as the content of oxidized proteins was not altered. Data obtained show the potential of this new methodology to be used in ecotoxicological studies.

16.P18. Microcystins affect antioxidant competence against peroxyl radicals in different organs of the fish Jenynsia multidentata (Jenyns, 1842) (Anablepidae)

16.P18. Microcystins affect antioxidant competence against peroxyl radicals in different organs of the fish Jenynsia multidentata (Jenyns, 1842) (Anablepidae)