Guilherme Malafaia

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 animals 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.

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 15xa0days. 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 1xa0h/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.

A pioneering study on cytotoxicity in Australian parakeets (Melopsittacus undulates) exposed to tannery effluent

Waste effluent from the tannery industry is a major source of environmental pollution. Considering that the bird intake of water contaminated with tannery effluent constitutes a potential genotoxic source, especially for birds inhabiting areas closest to tanning industries, the aim of this study is to assess the possible mutagenic effects that the intake may have on Melopsittacus undulatus (Australian parakeet). In order to do so, adult male and female M.xa0undulatus were distributed in two experimental groups: control (drinking water) and TE (5%). After 60 days of exposure, the micronucleus test, as well as tests looking for other nuclear abnormalities in the peripheral blood of the birds were performed. The male and female birds exposed to the pollutant have presented the highest total number of nuclear abnormalities, as well as increased individual abnormalities such as nuclei with symmetrical constricted bi-lobed/bi-nucleated erythrocytes, indented nuclei and micro-lobed nuclei (top)/micro-nuclei (bottom). In addition, the exposure to TE has caused a nuclear variant increase rarely reported in the literature concerning poultry erythrocyte nuclei. The birds exposed to the pollutant have presented the highest frequency of displaced nuclei forming different rotation/displacement angles within the cells. Therefore, the current study confirmed the toxicological potential of TE and was pioneer in showing that male and female M.xa0undulatus exposed to pollutant present the highest frequency of erythrocyte nuclear abnormalities, thus corroborating the initial hypothesis herein presented.

The genotoxicity and cytotoxicity of tannery effluent in bullfrog (Lithobates catesbeianus)

Some of the most polluting activities occur in bovine skin processing. Tannery generates effluents containing high concentrations of heavy metals and organic compounds. The phases composing the leather production process generate a large volume of tannery effluents that are often discarded in aquatic environments without any previous treatment. However, the effect these xenobiotics have on adult representatives belonging to the class Amphibia remains unknown. Thus, the aim of the present study is to assess the geno- and cytotoxic effects of tannery effluent on adult male bullfrogs (Lithobates castesbeianus) exposed to it. Accordingly, the animals were divided into the following groups: negative control (tannery effluent-free water), positive control (cyclophosphamide), and effluent (water added with 5% tannery effluent). The animals were euthanized for blood collection, and erythrocyte analyses were conducted after 35 and 90 days of exposure. The micronuclei (MN) frequency and the frequency of other nuclear abnormalities in each of the animals in the experimental groups were assessed in 2000 erythrocytes. According to the present results, the exposure to tannery effluents increased MN frequency as well as other nuclear abnormalities (i.e., lobed nuclei, binucleated cell, kidney-shaped nuclei, notched nuclei, and apoptotic cell) in the erythrocytes of animals in the effluent group and in the positive control group after 35 and 90 exposure days. Thus, the current study corroborated the hypothesis that the tannery effluent has aneugenic and clastogenic potential in adult male bullfrogs (L.xa0castesbeianus). The present study is the first to report such effect.

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 15xa0% concentrations diluted in water (period of 60xa0days). 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.

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.

Effects of abamectin on bullfrog tadpoles: insights on cytotoxicity

Abamectin is one of the most used pesticides worldwide. However, investigations about its effects on amphibian populations are rare. Thus, the present study sought to investigate possible cytotoxic effects on Lithobates catesbeianus tadpoles exposed to low abamectin concentrations diluted in water. Accordingly, four experimental groups were set: negative control, positive control (cyclophosphamide—40xa0mLxa0L−1), abamectin at concentrations 36xa0μgxa0a.i./L (ABA36 group), and 72xa0μgxa0a.i./L (ABA72 group), applied as Kraft® 36EC. The micronucleus test was conducted and other nuclear abnormalities in peripheral blood erythrocytes were checked after 24, 48, and 72xa0h of exposure to the treatments. The total of other nuclear abnormalities was influenced by the treatments, whereas the frequency of micronuclei was influenced by the exposure time. Such frequency was higher in the animals comprising the ABA72 group, which was assessed 72xa0h after the exposure had begun. The total of other nuclear abnormalities was influenced by the treatments. Animals in the positive control, ABA36, and ABA72 groups showed similar frequency of these abnormalities at 48 and 72xa0h. However, this frequency was statistically higher than that of animals in their respective negative control groups. Thus, the present study confirmed the hypothesis that the exposure of L. catesbeianus tadpoles to abamectin caused cytotoxic effects on them, although this exposure lasted short and the concentrations were low. It disclosed prospects for variations in the nucleus of erythrocytes circulating in amphibians, a fact that may provide an important/complementary approach for the detection of cytotoxicity caused by abamectin exposure.

Short-term social memory deficits in adult female mice exposed to tannery effluent and possible mechanism of action

The accumulated organic residues in tannery-plant courtyards are an eating attraction to small rodents; however, the contact of these animals with these residues may change their social behavior. Thus, the aim of the present study is to investigate whether the exposure to tannery effluent (TE) can damage the social recognition memory of female Swiss mice, as well as to assess whether vitamin C supplementation could provide information about how TE constituents can damage these animals memory. We have observed that resident females exposed to TE (without vitamin supplementation) did not explore the anogenital region, their body or chased intruding females for shorter time or with lower frequency during the retest session of the social recognition test, fact that indicates social recognition memory deficit in these animals. Such finding is reinforced by the confirmation that there was no change in the animals olfactory function during the buried food test, or locomotor changes in females exposed to the pollutant. Since no behavioral change was observed in the females exposed to TE and treated with vitamin C (before or after the exposure), it is possible saying that these social cognitive impairments seem to be directly related to the imbalance between the cellular production of reactive oxygen species and the counteracting antioxidant mechanisms (oxidative stress) in female mice exposed to the pollutant (without vitamin supplementation). Therefore, the present study evidences that the direct contact with tannery effluent, even for a short period-of-time, may cause short-term social memory deficits in adult female Swiss mice.

Protective effect of vitamin C in female Swiss mice dermally-exposed to the tannery effluent

Previous studies involving the oral exposure of mice to tannery effluents have found neurotoxic effects. However, studies about the effects the dermal exposure to pollutant have on the cognitive function of females have not been found in the literature. Thus, the aim of the current study is to investigate whether the dermal exposure of female Swiss mice to tannery effluents (2xa0h/day for 20 days) can cause cognitive impairment, as it was already evidenced in male Swiss mice. Furthermore, based on the administration of vitamin C (before or after the exposure to the xenobiotic), the current study also aims to assess the protective effect of vitamin C in female Swiss mice dermally-exposed to the tannery effluent. Female Swiss mice exposed to the tannery effluent (without vitamin supplementation) have shown lower novel object recognition index during the test session of the novel object recognition task, and they have descended significantly faster from the inhibitory avoidance platform when they were compared to mice belonging to the other groups, therefore evidencing memory deficit. However, the test performance of females receiving vitamin C was similar to that of control animals. Thus, the current study confirms the initial hypothesis that the dermal exposure to the pollutant, even for a short period, causes cognitive deficit in female Swiss mice. The herein presented findings also provide evidence that the mechanisms of action of the tannery effluent in these animals are related to oxidative damages in specific brain regions directed to the formation of short memory to perform aversive and object recognition tasks.