Behavioral and mutagenic biomarkers in tadpoles exposed to different abamectin concentrations.

It is known that pesticides such as abamectin (ABA) present cytotoxic effects on target organisms; however, the effects from ABA on non-target organisms such as amphibians are poorly understood. The aim of the current study is to investigate whether the exposure of Lithobates catesbeianus tadpoles to different abamectin concentrations [12.5, 25, and 50% of the median lethal concentration (LC50)] leads to behavioral and morphological changes and/or generates possible cytotoxic effects. The aggregation test showed that tadpoles exposed to the highest ABA concentrations did not respond to the stimulus from non-familial and unrelated co-specific species. On the other hand, there was no difference in the total number of crossings in the central line of the herein adopted apparatus between groups; it suggests that ABA did not affect animal locomotion in the aforementioned test, although changes in the normal swimming pattern of tadpoles exposed to the pesticide were recorded in the swimming activity test. In addition, the herein exposed animals did not respond to the predatory stimulus in the antipredator response test; this result suggests defensive response deficit caused by the pesticide. With respect to their oral morphology, tadpoles exposed to ABA presented the lowest scores for mandibular pigmentation and structures, as well as for dentition condition. Finally, it was possible seeing that the exposure to ABA, even at the lowest concentration (12.5% of the LC50), resulted in nuclear changes in the erythrocytes of the animals; these changes became evident in the increased number of micronuclei and in other nuclear abnormalities. Thus, besides confirming the cytotoxic potential of ABA in amphibians, the current study corroborates the hypothesis that the exposure to the herein investigated pesticide leads to behavioral and morphological changes in tadpoles, fact that may negatively reflect on the survival, as well as on natural populations of these individuals.

Impacts of tannery effluent on development and morphological characters in a neotropical tadpole.

Previous studies have shown the contamination of rivers and streams caused by tannery effluents (TE). However, the effects of TE on amphibians are still unknown. Thus, we evaluated whether the TE, even at low concentrations (0.2%), could cause asymmetric development (inferred through the fluctuating asymmetry indices (FA)) and changes in morphological characteristics of Physalaemus cuvieri larvae. Accordingly, the tadpoles were distributed in groups non-exposed (n=160) and exposed (n=160) to this TE, for 15days. The TE used presented a complex chemical composition (especially high concentrations of Cr, Cd and Ni and a diverse array of toxic organic compounds). We used three individual FA indices: FA1:=|R-L|, ii) FA2=|R-L|÷[(R+L)÷2] and FA3=(R-L)2, where "R" are "L" are the right and left sides, respectively. The larvae exposed to the TE had higher FA indexes (FA1, FA2 and FA3) for nostril-snout distance, relative to the control group larvae. Regarding eye-snout distance, asymmetry values were higher for the FA1 and FA2 indices (with no difference for the FA3 index), which were similar to those observed for the eye-width character. For eye length, FA2 was higher in animals of the effluent group. Tadpoles exposed to TE had significantly lower body masses than those in the control group, when controlling for body length and larval development stage. Furthermore, tadpoles in the effluent group had smaller body size and lower interocular distance. The frequency of tadpoles that showed any alteration in tail morphology as well as tail attachment position (dorsal and ventral) was significantly higher in the effluent group than in animals not exposed to the TE. Finally, our data show that exposure to TE delayed larval development of tadpoles. Thus, our pioneering study confirms the hypothesis that exposure of P. cuvieri to TE, at low concentrations, impacts development and morphological characters.

Mice exposure to tannery effluents changes their olfactory capacity, and their response to predators and to the inhibitory avoidance test.

The current study has assessed whether the oral and/or dermal exposure of C57Bl/6 J mice to tannery effluent (a complex pollutant consisting of xenobiotic mixtures) could damage their olfactory functions, as well as whether it changes their aversive behavior in the inhibitory avoidance test. Accordingly, the animals were distributed in groups which were exposed or not to this xenobiotic through two different routes (oral and dermal), for 15 days. The effluent group subjected to oral exposure received drinking water containing 5% tannery effluent, whereas the animals in the dermal group were exposed to raw tannery effluent for 1 h/day. The animals dermally exposed to the tannery effluent (males and females) have shown the highest latency to find palatable food in the buried food test. The shortest time spent by the animals (orally or dermally) exposed to tannery effluent in the safety zone of the apparatus used in the predator exposure test, as well as the longest time spent by them in the aversive zone, have shown failures in their perception to the risk represented by the presence of the predator (cat). The passive avoidance test results have shown that the dermal exposure to tannery effluent led to partial memory deficit in male and female mice; therefore, the present study has confirmed the tannery effluent toxicity to mammals. Moreover, the present study was pioneer in demonstrating that the dermal exposure to this xenobiotic, even for a short period-of-time, can change the olfactory and cognitive functions of animals, as well as lead to harmful consequences to their health.

Anxiety and memory deficits induced by tannery effluent in C57BL/6J female mice.

This study aimed to evaluate the behavior of female C57Bl/6J mice exposed to tannery effluents diluted in drinking water. Female mice were divided into a control group, in which the animals received only drinking water, and experimental groups, which received raw tannery effluent in 7.5 and 15 % concentrations diluted in water (period of 60 days). In the last experimental week, the mice (in diestrus phase) were subjected to different behavioral tests: elevated plus-maze, open-field test, forced swim test, and object recognition test. Our data demonstrated that exposure to tannery effluent increased the anxiety index of animals and decreased the locomotion ratio in the central quadrants/total, indicating an increase in anxiety-like behavior. Regarding the forced swim test, we did not observe changes in the evaluated behaviors. There were no statistically significant differences in the recognition index of the novel and familiar object in the groups exposed to tannery effluent compared with the control group, indicating a possible influence of the constituents of tannery effluent on cognition. Thus, our findings support the hypothesis that effluents, containing neurotoxic substances, could cause behavioral disruptions in female C57Bl/6J mice.

Memory deficit in Swiss mice exposed to tannery effluent.

Although it is known that tannery effluents constitute highly toxic pollutants whose effects in humans represent public health problems in several countries, studies involving experimental mammalian models are rare. In this context, the objective of the present study was to assess the effect of the exposure to tannery effluent on the memory of male and female Swiss mice. Animals of each sex were distributed into two experimental groups: the control group, in which the animals received only drinking water and the effluent group, in which the mice received 1% of gross tannery effluent diluted in water. The animals were exposed to the effluent by gavage, oral dosing, for 15days, ensuring the administration of 0.1mL of liquid (water or effluent)/10g of body weight/day. On the 14th and 15th experimental days the animals were submitted to the object recognition test. It was observed that the new object recognition indices calculated for the animals exposed to the effluent (males and females) were significantly lower than those obtained with the control group. The exposure to tannery effluent caused memory deficit in Swiss mice in a similar way for both sexes, reinforcing previous findings that these pollutants affect the central nervous system. It contributes to the knowledge in the area by attesting harmful effects to the cognition of such animals.

Predictive behaviors for anxiety and depression in female Wistar rats subjected to cafeteria diet and stress.

Obesity and chronic stress have been considered important public health problems that affect millions of people worldwide. Our aim was to analyze the effect of obesity associated with chronic stress on neurobehavioral parameters in female rats, considering that the association of these syndromes can enhance the negative effects on homeostasis. The animals were distributed into standard diet (Std), standard diet+stress (Std+stress), cafeteria diet (Cafe), and cafeteria diet+stress (Cafe+stress) groups. The animals of groups Std and Std+stress were fed with rodent standard feed. Groups Cafe and Cafe+stress, additionally to the standard feed, were offered palatable and calorie-rich processed food and cola-type soft drink ad libitum. From the eighth experimental week, groups Std+stress and Cafe+stress were subjected to restraint chronic stress model (50 days). After the stress protocol, predictive anxiety (open-field and elevated plus-maze tests) and depression (forced swim) were applied. The cafeteria diet was effective in inducing obesity. The ratio locomotion in the central quadrants/total locomotion evaluated during the open field test was not indicative of anxiogenic or anxiolytic effect in the animal's behavior. However, the elevated plus maze test showed that obese and stressed animals were prone to higher anxiety levels. In addition, the obese and stressed animals display less climbing behavior than all the other groups, which can be considered an indicator of depression-like behavior. Nevertheless, it is suggested that the mechanisms involved in effects of obesity associated with chronic stress be better investigated in female rats, considering the organic complexity related to these modern illnesses.