Computer-based inquiry to enhance science process skills in undergraduate students using physiological data.

The analysis of spontaneous tail coiling (STC) in zebrafish embryos is a functional parameter that allows the study of motor development. It has recently gained relevance as a biomarker to assess the neurotoxicity of environmental substances. Its practicability in the laboratory makes it an excellent pedagogical tool for promoting students' inquiry skills. However, the time and cost of materials and facilities limit their usage in undergraduate laboratories. This study presents the design of a computer-based educational module called ZebraSTMe, which is based on a tail coiling assay and aims to improve science process skills in undergraduate students by connecting them to relevant and novel content. We evaluate students' perception of learning, the quality of materials used, and the knowledge gained. Our results show that students perceived an improvement in their statistical analysis, representation, and discussion of experimental data. Additionally, the students evaluated the quality and ease of use of the materials and provided feedback for revision. A thematic analysis of the opinions revealed that the module activities promoted students' reflection on their professional strengths and weaknesses.NEW & NOTEWORTHY ZebraSTMe is a novel computer-based educational module that utilizes spontaneous tail coiling analysis in zebrafish embryos to enhance undergraduate students' scientific inquiry skills. By addressing the challenges of time, cost, and laboratory resources, the module improves students' science process skills and promotes reflection on their professional strengths and weaknesses. The innovative ZebraSTMe exemplifies the potential of integrating cutting-edge research topics into undergraduate education, leading to more engaging and effective learning experiences in physiology and other scientific disciplines.

Exposure to low doses of inorganic arsenic induces transgenerational changes on behavioral and epigenetic markers in zebrafish (Danio rerio).

The ability of environmental pollutants to alter the epigenome with resultant development of behavioral alterations has received more attention in recent years. These alterations can be transmitted and affect later generations that have not been directly in contact with the contaminant. Arsenic (As) is a neurotoxicant and potent epigenetic disruptor that is widespread in the environment; however, the precise potential of As to produce transgenerational effects is unknown. Our study focused on the possible transgenerational effects on behavior by ancestral exposure to doses relevant to the environment of As, and the epigenetic mechanisms that could be involved. Embryos of F0 (ancestral generation) were directly exposed to 50 or 500 ppb of As for 150 days. F0 adults were raised to produce the F1 generation (intergeneration) and subsequently the F2 generation (transgeneration). We evaluated motor and cognitive behavior, neurodevelopment-related genes, and epigenetic markers on the F0 and F2 generation. As proposed in our hypothesis, ancestral arsenic exposure altered motor activity through the development and increased anxiety-like behaviors which were transmitted to the F2 generation. Additionally, we found a reduction in brain-derived neurotrophic factor expression between the F0 and F2 generation, and an increase in methylation on histone H3K4me3 in the nervous system.

Tryptophan supplementation helps totoaba (Totoaba macdonaldi) juveniles to regain homeostasis in high-density culture conditions.

High-density culture brings with it chronic stress situations that affect fish welfare. In order to evaluate the effect of tryptophan (Trp) levels on the response to stress, Totoaba macdonaldi juveniles were stocked at low (13.5 kg m-3) and high (27.0 kg m-3) densities (32.5 and 56.4 kg m-3, respectively, at the end of the experiment) in 100-L tanks and fed for 63 days with experimental diets containing different Trp levels: control diet CD0.42 (0.42%) and three supplemented diets with 0.99, 1.55 and 2.19% (0.99Trp, 1.55Trp and 2.19Trp, respectively) (three tanks × density × diet). The high-density stocking fed with CD0.42 diets showed significantly increased blood parameters. Trp decreased catalase (CAT) activity in low- and high-density stocking, while the superoxide dismutase (SOD) activity showed no difference. Serotonin (5-hydroxytryptamine, 5-HT) content decreased, and the serotonin turnover ratio (5-HIAA:5-HT) increased in the brains of fish fed with the CD0.42 diet. Indeed, Trp-supplemented diets helped to restore homeostasis in high-density growth conditions as evaluated by the hematological and plasma parameters as well as the serotonergic activity. When the fish were provided a diet containing moderate Trp levels, plasma cortisol increased under high-density conditions. However, no differences were observed among stock densities when totoaba were fed with the 2.19Trp diet. Notably, survival was unaffected by both Trp or densities, but weight gain (WG) decreased with the dietary Trp levels in the high density culture. In sum, Trp supplementation decreased the parameter values linked to stress response on totoaba juveniles cultured at high stock densities.

Quantification of Spontaneous Tail Movement in Zebrafish Embryos Using a Novel Open-Source MATLAB Application.

Recently, the evaluation of spontaneous tail movement (STM) in zebrafish embryos has been proposed as a possible neurotoxicity marker in studies on behavioral changes due to chemical exposure. Currently, automated STM quantification can only be performed using high-cost commercial software. We have developed a simple and reliable MATLAB® script called ZebraSTM that allows the automatic analysis of STM in multiple zebrafish embryos.

Neurobehavioral Alterations in Zebrafish Due to Long-Term Exposure to Low Doses of Inorganic Arsenic.

Inorganic arsenic (As) is one of the most ubiquitous and toxic substances with widespread health effects on human populations and biodiversity. Although arsenic is a frequent surface water pollutant, there is scant evidence about neurotoxicity in aquatic species in different stages of development. In the present study, we investigated the neurobehavioral effects of chronic exposure to environmentally relevant doses of arsenic. We exposed zebrafish to 50 and 500 ppb during the larval, juvenile, and adult stage (from 4 h to 150 days postfertilization). We then used broad behavioral screening to evaluate motor function, social behavior, learning and memory, and anxiety-like behaviors. Our results show that arsenic exposure to 500 ppb alters motor function from the embryo to the adult stage. Furthermore, during the adult phase, associative learning and the sensorimotor response are affected with both high and low doses of As, respectively. Notably, exposure to 500 ppb of As induces behaviors associated with anxiety, during the juvenile and adult phase but not the larval stage, without changes in whole-body cortisol levels. These results indicate that chronic exposure to arsenic during their lifespan is capable of producing alterations in different behavioral markers in aquatic vertebrates.

Evidence for effects on thermoregulation after acute oral exposure to type I and type II pyrethroids in infant rats.

Most pyrethroid (PYR) insecticides may be classified either as type-I compounds, which produce whole body tremors and hyperthermia, or type-II compounds, which produce salivation, choreoathetosis, and hypothermia (i.e., producing T and CS neurobehavioral syndromes, respectively). This classification is based on clinical observations in adult rats and mice after intracerebroventricular or intravascular administration of highly effective acute (bolus) doses. PYR neurotoxicity in infant animals is not characterized as much as in adult animals. Endpoints informing on vital determinants of mammal's maturation, such as body temperature may help recognizing age-related differences in susceptibility to PYRs. In this work, body temperature (Tb) was monitored at 30-min intervals after acute oral exposure to T-syndrome PYR bifenthrin (BIF), CS-syndrome PYR cypermethrin (CYPM), and a BIF­CYPM mixture in weanling rats by using a subcutaneous temperature monitoring system. In both single-compound assays, a time- and dose-related decline of Tb was the most evident impact on thermoregulation observed starting at ~2­3 h after dosing.Moreover, 15­18 mg/kg BIF induced a mild increase in Tb before the hypothermic action was apparent. The lowest effective dose for temperature perturbation was 15mg/kg for BIF and 10mg/kg for CYPM, and moderate neurobehavioral alterations were evident at 12 and 10mg/kg, respectively. When low effective doses of BIF and CYPM were co-administered mild behavioral effects and a transient increase in Tb (p=0.02) were observed at 1­2 h, and no Tb decline was apparent afterwards compared to control animals. Noteworthy, the hypothermic action of BIF in infant rats was quite different from the hyperthermia consistently reported in studies using mature animals. Our results suggest that body temperature monitoring may be useful as a complementary assessment to reveal qualitative age-specific pesticide effects in rats.

Atrazine is primarily responsible for the toxicity of long-term exposure to a combination of atrazine and inorganic arsenic in the nigrostriatal system of the albino rat.

Chronic and simultaneous exposure to a variety of chemicals present in the environment is an unavoidable fact. However, given the complexity of studying chemical mixtures, most toxicological studies have focused on the effects of short-term exposure to single substances. The aim of this study was to evaluate the effects on the nigrostriatal system of the chronic, simultaneous exposure to two widely distributed substances that have been identified as potential dopaminergic system toxicants, inorganic arsenic (iAs) and atrazine (ATR). Six groups of rats were treated daily for one year with atrazine (10mg ATR/kg), inorganic arsenic (0.5 or 50mgiAs/L of drinking water), or a combination of ATR+0.5mgiAs/L or ATR+50mgiAs/L. The 50mgiAs/L group showed locomotor hypoactivity, while all treatments decreased motor coordination in contrast no effects of treatment were found on the place and response learning tasks. Regarding markers for liver and muscle damage, there were no differences between groups in creatine kinase (CK) or aspartate transaminase (AST) activities, while decreases in lactate dehydrogenase (LDH) levels were found in some exposed groups. The striatal DA content was significantly reduced in ATR, 0.5mgiAs/L, ATR+0.5mgiAs/L, and ATR+50mgiAs/L groups, in comparison to the control group. The number of mesencephalic tyrosine hydroxylase positive cells decreased in the ATR and ATR+0.5mgiAs/L groups compared to the control. In contrast, immunoreactivity to cytochrome oxidase was reduced compared to the control in all treated groups, except for the group treated with 0.5iAsmg alone. Our results indicate that ATR has deleterious effects on dopaminergic neurons and that the combination of ATR and iAs does not exacerbate these effects.

Chronic atrazine exposure causes disruption of the spontaneous locomotor activity and alters the striatal dopaminergic system of the male Sprague-Dawley rat.

The herbicide atrazine (ATR) is widely used around the world, and is a potential toxicant of the dopaminergic systems. Nigrostriatal and mesolimbic systems are the two major dopaminergic pathways of the central nervous system; they play key roles mediating a wide array of critical motor and cognitive functions. We evaluated the effects of exposing male rats for one year to 10 mg ATR/kg B.W. on these systems using motor and cognitive tasks and measuring monoamine content in the striatum, nucleus accumbens, prefrontal cortex, and hypothalamus. ATR administration resulted in impaired motor coordination and greater spontaneous locomotor activity only after 10 to 12 months of exposure. Chronic exposure to 10 mg ATR decreased striatal dopamine, but had no effect on accumbal, hypothalamic or cortical monoamine content. Chronic ATR exposure caused discrete changes in learning tasks that involve either the striatum or the nucleus accumbens. These results indicate that chronic exposure to ATR preferentially targets the nigrostriatal dopaminergic pathway, in comparison to the other dopaminergic pathways evaluated in this study, inducing behavioral and neurochemical alterations. In order to unveil the full extent of atrazine's effects on the nervous system, other neurochemical systems should be considered in future studies.