Caracterização comportamental e distribuição de neurônios inibitórios em um modelo animal de autismo induzido por ácido valpróico

Autism comprises a heterogeneous group of neurodevelopmental disorders that affects the brain maturation and produces sensorial, motor, language and social interaction deficits in early childhood. Several studies have shown a major involvement of genetic factors leading to a predisposition to autism, which are possibly affected by environmental modulators during embryonic and post-natal life. Recent studies in animal models indicate that alterations in epigenetic control during development can generate neuronal maturation disturbances and produce a hyper-excitable circuit, resulting in typical symptoms of autism. In the animal model of autism induced by valproic acid (VPA) during rat pregnancy, behavioral, electrophysiological and cellular alterations have been reported which can also be observed in patients with autism. However, only a few studies have correlated behavioral alterations with the supposed neuronal hyper-excitability in this model. The aim of this project was to generate an animal model of autism by pre-natal exposure to VPA and evaluate the early post-natal development and pre-puberal (PND30) behavior in the offspring. Furthermore, we quantified the parvalbumin-positive neuronal distribution in the medial prefrontal cortex and Purkinje cells in the cerebellum of VPA animals. Our results show that VPA treatment induced developmental alterations, which were observed in behavioral changes as compared to vehicle-treated controls. VPA animals showed clear behavioral abnormalities such as hyperlocomotion, prolonged stereotipies and reduced social interaction with an unfamiliar mate. Cellular quantification revealed a decrease in the number of parvalbumin-positive interneurons in the anterior cingulate cortex and in the prelimbic cortex of the mPFC, suggesting an excitatory/inhibitory unbalance in this animal model of autism. Moreover, we also observed that the neuronal reduction occurred mainly in the cortical layers II/III and V/VI. We did not detect any change in the density of Purkinje neurons in the Crus I region of the cerebellar cortex. Together, our results strengthens the face validity of the VPA model in rats and shed light on specific changes in the inhibitory circuitry of the prefrontal cortex in this autism model. Further studies should address the challenges to clarify particular electrophysiological correlates of the cellular alterations in order to better understand the behavioral dysfunctions

Atividades comportamentais e comportamento alimentar de rã touro, Lithobates catesbeianus (Shaw, 1802), de pigmentação normal e albina em cativeiro

The behavior of bullfrogs reared in captivity must be well understood to support management practices that use efficient feeding regimes. In general, bullfrogs reared in captivity have normal pigmentation, but to develop an enhanced product, some studies have investigated the introduction of albino individuals in frog farms. The present study characterized the behavior of both pigmented and albino bullfrogs reared in captivity. In an initial experiment, 48 bullfrogs (70.5 ± 25.6 g) housed in small stalls were fed once a day at random times. Frogs were filmed and the images showed that both the pigmented and albino varieties behaved similarly: food intake was more frequent at dawn followed by light periods; moving and resting in dry areas may be associated to feeding events; frogs appeared to anticipate feeding time and to rest in the water more frequently in periods other than feeding time; daylight is the recommended period for feeding both pigmented and albino frogs. In a second experiment, 72 albino bullfrogs were fed at fixed times (10 a.m. or 4 p.m.) in small stalls. An initial weight of 23.8 ± 7.6 g was considered to evaluate frog performance, and after the animals reached 60.0 ± 20.0 g, they were filmed for behavior analyses. Food intake varied as a function of feeding time, and frogs were more likely to eat during the early hours of the day and immediately after receiving fresh food. Frogs fed only in the afternoon changed their behavior. Food supplied twice a day stimulated the albino frogs to eat at different times, but did not increase growth. Although fresh food stimulated feeding behavior, food intake was more frequent at dawn. Food supplied at this time of day should therefore be further investigated. The results did not indicate a more suitable feeding time (10 a.m. or 4 p.m.) for albino bullfrogs, or any advantage in using two feedings per day. The results provide xvi important information about bullfrogs in terms of food supply regime and activity preferences throughout the day. This novel information will contribute to future studies in this area

Avaliação comportamental e neuroquímica de ratos em dessincronização forçada: possíveis implicações para um modelo animal de oscilações no humor

Bipolar disorder has been growing in several countries. It is a disease with high mortality and has been responsible by the social isolation of the patients. Bipolar patients have alterations in circadian timing system, showing a phase shift in various physiological variables. There are several arguments demonstrating alterations in circadian rhythms may be part of the bipolar disorder pathophysiology. Given the necessity for further elucidation, the goal of this study was to validate the forced desynchronization protocol as an animal model for bipolar disorder. To do this, Wistar rats were submitted to a forced desynchronization protocol which consists in a symmetrical light dark cycle with 22h. Under this protocol, rats dissociate the locomotor activity rhythm into two components: one synchronized to the light / dark cycle with 22h, and another component with period longer than 24 hours following the animal endogenous period. These rhythms with different periods sometimes there is coincidence, which we named CAP (Coincidence Active Phase) and the opposite phase, non-coincidence, called NCAP (Non-Concidence Active Phase). The hypothesis is that in CAP animals present a mania-like behavior and animals in NCAP depressive-like behavior. We found some evidence described in detail throughout this thesis. In sum, the animals under forced desynchronization protocol were more stressed, showed an increase in stereotypic behaviors such as grooming and reduction in other behaviors such as risk assessment and vertical exploration when compared to the control group. The CAP animals showed increased locomotor activity, especially during the dark phase when compared to controls (rats under T24) and less depressive behavior in the forced swim test. The animals in NCAP showed a higher anxiety in elevated plus maze, but they don t have ahnedonia. The animals under dissociation have more labeled 5HT1A cells at the amygdala area, which appoint that they have more amygdala inhibition. Taking these data together, we could partially validated the forced desynchronization protocol as an animal model for mood oscillations

Boto cinza (Sotalia guianensis, Van Benédén, 1864) (Cetacea, Delphinidae): Atividade aérea, forrageio e interações inter-específicas, na Praia de Pipa (Tibau do Sul RN) e estudo comparativo entre duas populações do Nordeste do Brasil

Sotalia guianensis is a small cetacean of the Delphinidae family, with coastal habits and whose area of distribution ranges from Florianópolis (27º35'S, 48º34'W), in Brazil, to Honduras (15º58'N, 85º42'W). At Pipa beach, on the south coast of RN state, in Brazil, the species occur throughout the year. The present study was carried out in two bays, which are bordered by cliffs. The animals were monitored from vantage points, using the "Ad libitum" and "all the occurrences" methods; during the years of 1999 and 2004. The study was divided in 4 chapters: Behavioral standards of two populations of gray dolphin, (Sotalia guianensis, Van Benédén, 1864) in the northeast of Brazil; Aerial activity of the gray dolphin: its possible function and the influence of environmental and behavioral factors; The influence of daily and monthly variation of the tides, of the period of the day and group size on the gray dolphin forage activity; kleptoparasitism interactions of frigatebird (Fregata magnificens, Mattheus, 1914) during the gray dolphin forage activity. The results have shown that the gray dolphin has a varied and complex behavioral repertoire. The leap is the most frequent behavior; the aerial activity is diffuse during daylight and is influenced by some factors, such as the level of the tide and social factors. The gray dolphin, when in the bay, most frequently feeds isolate or in small groups. The forage is diffuse during daylight; however, being more frequent in the morning and is influenced by the daily and monthly variation of the tide. At Pipa beach, kleptoparasitarian interactions were registered between the gray dolphin and the frigatebird (Fregata magnificens). The frigatebird forage strategy consists basically of two ways: to fly over great extensions searching for dead fish and to steal food (kleptoparasitism). These interactions were predominantly carried out between immature and female adult birds and adult and immature dolphins, and occurred during daylight. The present study can be considered an initial landmark to a better knowledge on the gray dolphin surface behavior, especially regarding the aerial behavioral repertoire and forage strategy of this species. However, it is necessary to continue these studies, so that we can understand better the complex social life of these animals and thus create effective measures for its conservation

Padrão de atividades e de interações sociais de animais juvenis em dois grupos selvagens de Callithrix jacchus

The juvenile period represents the developmental phase between weaning and sexual maturity. Weaning occurs when the youngster does not receive direct care from the caretakers anymore. Individuals in the species Callithrix jacchus live in groups composed by the reproductive pair and successive twin sets. Cooperative care is the rule. Infants are weaned early, and from then on, food is provided by the adults in the group. These animals present high levels of social interactions, through play, grooming and social contact. During infant age, the twin becomes the main partner. There are few studies about the juvenile period, especially on Callithrix gender. The objective of this study was describing the pattern of activities and social interactions of four sets (one single and three twin sets) during juvenile phase in two Callithrix jacchus groups. We used instantaneous and continuous focal sampling for juveniles and scan sampling for adults behavioral recordings. Juveniles presented the same behavioral pattern as the adults relating the activity budget, in particular, foraging along the months. The composition of the diet was the same as that of the adults. Food transfer ended along the juvenile period. Social play as much as grooming were important socializing activities for the juveniles. The young individuals in the group were the main partners in social play, specially the twin. Adults were the main partners in grooming interactions. Scent marking differed between twins in the male/female sets, the female presenting the highest levels of marking. The juveniles were independent from adults in foraging activity. Social interaction varied according to group composition, but in general, interacted more with the twin and with the youngsters (infants and subadults), except in grooming. Even presenting many similarities, juveniles showed some differences between genders, which indicates the differentiation in behavior towards reproductive strategies early in the juvenile period

Aprendizado espacial-temporal em um ambiente complexo sem restrição alimentar em ratos

Timeplace learning is the capacity of organisms to associate both space and time with a biological relevant stimulus such as food. Experiments are usually done with food restricted animals due to the belief that food system activation is necessary for timeplace learning. Another line of thought suggest that, in addition to food system activation, response cost should be increased to effectively allow timeplace discrimination. The purpose of this experiment was to test whether a complex environment, which presumably implied in a heightened response cost, would facilitate timeplace association in satiated rats using a highly palatable food as reward. Nine rats were trained in a timeplace task for 30 nonconsecutive days. A large experimental box (1x1m) divided in four compartments was used. To access each compartment the animal had to overcome a series of obstacles such as ramps, staircases and mazes. Two feeders localized in opposite compartments were rewarded with sunflower seeds in two daily sessions. One feeder offered the reward during the morning sessions while the second feeder in afternoon sessions. After the 15th day of training, the animals began to show a preference for the correct feeder during the correct time of day expressed by increased frequency of visits as well as lower latency to access the feeders. These results suggest that satiated animals are also capable of learning a timespace task as far as the experimental context is complex enough to result in a higher response cost

Efeitos da alteração dos níveis de ansiedade no desempenho de ratos em uma tarefa de discriminação apetitiva

The plus-maze discriminative avoidance paradigm has been used to study the relationship between aversive memory and anxiety. The present study aims to verify if the elevated plus-maze can provide information about appetitive memory and anxiety, through a task motivated by food reward. Animals were allowed to explore an elevated plus-maze and received reinforcement in one of the enclosed arms. In a test session performed 24h later, in the absence of reward, rats showed preference for the previously rewarded enclosed arm over the neutral enclosed arm. The administration of diazepam and pentylenetetrazole before training induced, respectively, anxiolytic and anxiogenic effects (as evaluated by open-arm exploration). Both drugs induced amnestic effects, i.e., lack of preference for the rewarded arm in the test session. The results suggest that appetitive memory can be influenced by anxiety levels as well. The plus-maze appetitive discrimination task seems to be a useful model to investigate the relationship between memory and anxiety

Efeitos de vocalizações de co-específicos e do escuro sobre o ritmo circadiano da atividade motora em sagüis (Callithrix jacchus)

The principal zeitgeber for most of species is the light-dark photocycle (LD), though other environment factors as food availability, temperature and social cues may act. Daily adjustment of the circadian pacemaker may result from integration of environmental photic and non-photic cues with homeostatic cues. Characterization of non-photic effects on circadian timing system in diurnal mammals is scarce in relation to nocturnal, especially for ecologically significant cues. Thus, we analyzed the effect of conspecific vocalizations and darkness on circadian activity rhythm (CAR) in the diurnal primate Callithirx jacchus. With this objective 7 male adults were isolated in a room with controlled illumination, temperature (26,8 ± 0,2°C) and humidity (81,6 ± 3,6%), and partial acoustic isolation. Initially they were under LD 12:12 (~300:2 lux), and subsequently under constant illumination (~2 lux). Two pulses of conspecific vocalizations were applied in total darkness, separated by 22 days, at 7:30 h (external time) during 1 h. They induced phase delays at circadian times (CTs) 1 and 10 and predominantly phase advances at CTs 9 and 15. After that, two dark pulses were applied, separated by 14 days, during 1 h at 7:30 h (external time). These pulses induced phase delays at CTs 2, 3 and 18, predominantly phase advances at CTs 8, 10 and 19, and no change at CT 14. However, marmosets CAR showed oscillations in endogenous period and active phase duration influenced by vocalizations from animals outside the experimental room, which interfered on the phase responses to pulses. Furthermore, social masking and relative coordination with colony were observed. Therefore, phase responses obtained in this work cannot be attributed only to pulses. Afterwards, pulses of conspecific vocalizations were applied in total darkness at 19:00 h (external time), during 1 h for 5 consecutive days, and after 21 days, for 30 consecutive days, on attempt to synchronize the CAR. No animal was synchronized by these daily pulses, although oscillations in endogenous period were observed for all. This result may be due to habituation. Other possibility is the absence of social significance of the vocalizations for the animals due to random reproduction, since each vocalization has a function that could be lost by a mixture of sounds. In conclusion, conspecific vocalizations induce social masking and relative coordination in marmosets CAR, acting as weak zeitgeber

Estudo da preferência manual durante o desenvolvimento ontogenético em sagüi comum ( Callithix jacchus)

The study of manual preference is a widely used approach to investigate cerebral laterality in nonhuman primates. However, in New World primates, little is known about the ontogenesis of hand use asymmetry, in both forced and spontaneous activities, as well as how they correlate with sexual hormones. Accordingly, a longitudinal study was conducted on the manual preference of 6 female and 4 male common marmosets (Callithrix jacchus). The study included the record of forced tests to reach for the food using only one hand (forced activity) and activities such as grooming (auto and social), scratching, grasping the food and hanging, in weekly sessions from the infantile (4 months) to the early adult phase (15 months). Feces samples were also collected, at least once a week, to evaluate the level of gonadal steroids and their influence on these behaviors. In the forced activity, the results confirm the influence of the development period on manual preference during feeding, shown by the increase in lateral stability when grasping the food between the juvenile and adult phases. During this period, a sexual hormone effect on development was also observed, mainly of progesterone in females and androgen in males, but no difference between sexes was found. In the females, progesterone also influenced the manual preference index, with a proportional increase in the degree of manual asymmetry during puberty. With respect to spontaneous activities, the animals showed proportional use of both hands when scratching, hanging, holding the food and grooming. A positive correlation was also found between the preference for holding the food in forced activities and in spontaneous activities

O gato doméstico (Felis catus) responde à sinais gestuais? possíveis implicações do convívio social

The cats (Felis catus) were domesticated about 9,500 years ago due to the advent of agriculture, being used to control the pests that devastated the food harvested. These animals went through an artificial selection and over generations and millennia had their behavior and morphology changed by humans. This process of domestication by man gave rise to a special ability, the understanding of human pointing gestures, clearly noticed while we feed our pets. Our goal in this study was to assess the comprehension of pointing gestures by cats and also verify the influence that social interactions exerts on the development of this ability. We found that experimental subjects from both groups, solitary animals and social animals, were able to follow human indication in order to find hidden food. However, social interaction had no effect on cats performances. The ability tested here probably evolved during the process of domestication of this species, and social interaction seems to exert little or no influence upon its expression

Comportamento alimentar do camarão marinho farfantenaeus subtilis em condições laboratoriais

Shrimp culture represents an important activity to brazilian economy. The northeastern region has presented high levels of production because of its climatic conditions. An important factor for the activity´s major development is related to the introduction of the species Litopenaeus vannamei. The use of an exotic species can disturb the ecosystem. In the last decades, L. vannamei has been the only species cultivated in Brazilian farms, there not being an alternative species for shrimp culture. So, there is an urgent need to developing new studies with the native species, which might represent an alternative concerning shrimp production, with emphasis on Farfantepenaeus subtilis. Another important aspect related to the activity is feeding management, once it is quite usual that feed offer on the pond does not take into account either the species´ physiology and behavior or the influence of environmental variables, such as light cycle and substrate. That knowledge may optimize management and so reduce the impact of effluents in the environment. This study´s objective was characterizing feeding behavior of F. subtilis in laboratory. For that, an ethogram was developed, using 20 wild animals which were observed through ad libitum and all occurrences methods. Two experiments were developed in order to register feeding behavior on different substrates, along 15 days, each. In the first experiment, 40 animals were distributed in eight aquaria, half being observed during the light phase of the 24 hour cycle and the other half in the dark phase, both in halimeda substrate. In the second experiment, 20 animals were distributed in four aquaria, under similar conditions as the previous ones, but in sand substrate. In both experiments, animals were observed respectively one, four, seven and ten hours after the beginning of the phase, for light phase, for the dark phase, in ten minute observation windows, before and immediately after feed offer. The following behaviors were registered: feed ingestion, ingestion of other items, inactivity, exploration, vertical exploration, swimming, crawling, digging, burrowing, and moving by the animals. Observation windows after feed offer also included latency to reach the tray and to ingest feed. Nineteen behaviors were described for the species. F. subtilis presented more behavioral activities in halimeda substrate even in the light phase, while burrowing was predominant in sand substrate. In both substrates, moving, crawling and exploration were more frequent after feed offer, but inactivity and burrowing were more frequent before that. Feed ingestion was more frequent in halimeda, both in light and dark phases. Weight gain was also more prominent in that substrate. In sand substrtate, ingestion was more frequent in the dark phase, which suggests that higher granulometry facilitates feed ingestion in F. subtilis juveniles. Our results demonstrate the importance of studies for the better knowledge of the species, specially its response to environmental stimuli, in order to improve animal management

O papel das rotas e da obtenção de informações sobre a eficiência no forrageio de dinoponera quadriceps em ambiente natural

In this study, we investigated the role of routes and information attainment for the queenless ant species Dinoponera quadriceps foraging efficiency. Two queenless ant colonies were observed in an area of Atlantic secondary Forest at the FLONA-ICMBio of Nisia Floresta, in the state of Rio Grande do Norte, northeastern Brazil, at least once a week. In the first stage of the study, we observed the workers, from leaving until returning to the colony. In the second stage, we introduced a acrylic plate (100 x 30 x 0,8 cm) on a selected entrance of the nest early in the morning before the ants left the nest. All behavioral recordings were done through focal time and all occurence samplings. The recording windows were of 15 minutes with 1 minute interval, and 5 minute intervals between each observation window. Foraging was the main activity when the workers were outside the nest. There was a positive correlation between time outside the nest and distance travelled by the ants. These variables influenced the proportion of resource that was taken to the nest, that is, the bigger its proportion, the longer the time outside and distance travelled during the search. That proportion also influenced the time the worker remained in the nest before a new trip, the bigger the proportion of the item, the shorter was the time in the nest. During all the study, workers showed fidelity to the route and to the sectors in the home range, even when the screen was in the ant´s way, once they deviated and kept the route. The features of foraging concerning time, distance, route and flexibility to go astray by the workers indicate that decisions are made by each individual and are optimal in terms of a cost-benefit relation. The strategy chosen by queenless ants fits the central place foraging and marginal value theorem theories and demonstrate its flexibility to new informations. This indicates that the workers can learn new environmental landmarks to guide their routes

Uso de rotas na área de uso e a relação com comportamento alimentar em Callithrix jacchus

Callithrix jacchus are small primates that live in cooperative reproductive family groups. They explore their home range in search of fruits, exudates and animal prey. In this study we investigate the existence of traveling routes and its relation with the feeding habits in a group of Callithrix jacchus. The group was followed for 6 months in an area of Atlantic secondary Forest at the FLONA-ICMBio of Nísia Floresta, Rio Grande do Norte. Two observers in alternated days collected data referring to the group s location using a GPS navigation device, plotting data in 5 minute intervals, and with a position accuracy under 9 meters. All behavioral recordings were done through focal time samplings. The recording windows were 15 minutes with 1 minute intervals. The main activity was foraging, which propelled the animals to explore the environment with inconsistent intensity through the months, and correlated with the location of fruits, exudates and sleeping sites. From another standpoint, most activities were focused in the core areas that featured most sleeping sites, exudates trees and fruit trees. Insects, mostly Orthopterans, were hunted in all areas. The greater ratio of movement was registered during the last hours of sunlight, when animals returned to the sleeping sites and ate a greater number of fruits. The spatial and seasonal distribution of fruits forced the animals to travel long routes. The capacity to remember the location and navigate efficiently through feeding sources is important to save energy and time costs. Learning and familiarizing with the environment through the use of landmarks and acquisition of new information is extremely important to increase the chances of survival in a constantly changing environment

O efeito da dessincronização circadiana sobre o metabolismo

The lost of phase relationship between rhythms and behaviour can, and often do, undesirable consequences. The purpose os study was to ascertain the effect of circadian desynchronization in T22 about metabolism of wistar rats. The subjects consisted of 24 animals separated in two groups: control (n=12) T24 with 8 weeks of aged and experimental group (n=12) T22, also with 8 weeks of aged. Both the groups were subject to register of locomotor actitivity, body temperature, body weight and food intake in all the experiment. And more, both the groups were subject to food deprivation, running in treadmill and forced swimming. The results show rhythm of locomotor activity and body temperature desynchronized. Dont exist diference in body weight between both the groups (T24 = 386,75±40,78g e T22 380,83±44,28g) . However, the food intake was different between the phases, light and dark, in intergroup and intragroup. The body temperature was not different in food deprivation. The same ocurred for running in treadmill and forced swimming. Since similar alterations occur in shift workers, it is proposed that the experimental paradigm presented in this manuscript is a useful model of shift work. That is, alterations in activity/rest cycles and consummatory behavior can affect the health of organism