Influência da salinidade e temperatura da água nas respotas comportamental e fisiológica de camarões marinhos Litopenaeus vannamei(BOONE 1931)

The shrimp Litopenaeus vannamei has been grown in highly variable environments, especially in relation to salinity and water temperature. The adjustment to such conditions mainly involves changes in behavior, physiology, particularly in the immune response. This may consequently reduce the welfare of these animals. Despite the widespread farming of the species, little is known about their behavioral and physiological responses under stressful conditions. Thus, the objective of this study was to assess the influence of different salinities and temperatures in the behavior of the marine shrimp L. vannamei, and its relation to the total hemocytes count. In the laboratory, juvenile shrimp were kept in glass aquaria with a closed water recirculation system, continuous aeration and filtration, and under a 12:12 h light/dark cycle. Behavioral observations occurred 1, 4, 7 and 10 h after the start of each phase (light or dark). To assess the influence of salinity, shrimp were first acclimated and then observed at 2, 30 or 50 ppm salinity water, while temperatures tested were 18, 28 and 33 ° C. At the end of each experiment (30 days), shrimp hemolymph was collected for subsequent total hemocytes count (THC), a parameter used to assess stress. In general, feeding behavior was modified under lower salinity and temperature, with reduced values in feeding, exploration and digestive tract filling. Inactivity and burrowing were prevalent under extreme conditions water salinity and temperature, respectively: 2 and 50 ppm and 18 and 33 ° C; crawling was also less frequent under these conditions. In regards to light/dark cycle, shrimp were more active during the dark phase (crawling and swimming), while burrowing was higher during the light phase, regardless of salinity or temperature of the water. Inactivity behavior did not vary according to the light/dark cycle. Moreover, the total hemocytes count (THC) was reduced under 2 and 50 ppm salinity and 18 ° C temperature. Farming of L. vannamei under extremely low or high salinities and low temperatures is harmful. This suggests the species must be cultivated in salinities closer to those of the sea as well as at high temperatures, which seems to be ideal for a management focused on animal welfare, therefore, producing healthier shrimp

Estudo da organização funcional do sistema circadiano por meio de ferramentas computacionais e matemáticas

Circadian rhythms are variations in physiological processes that help living beings to adapt to environmental cycles. These rhythms are generated and are synchronized to the dark light cycle through the suprachiasmatic nucleus. The integrity of circadian rhythmicity has great implication on human health. Currently it is known that disturbances in circadian rhythms are related to some problems of today such as obesity, propensity for certain types of cancer and mental disorders for example. The circadian rhythmicity can be studied through experiments with animal models and in humans directly. In this work we use computational models to gather experimental results from the literature and explain the results of our laboratory. Another focus of this study was to analyze data rhythms of activity and rest obtained experimentally. Here we made a review on the use of variables used to analyze these data and finally propose an update on how to calculate these variables. Our models were able to reproduce the main experimental results in the literature and provided explanations for the results of experiments performed in our laboratory. The new variables used to analyze the rhythm of activity and rest in humans were more efficient to describe the fragmentation and synchronization of this rhythm. Therefore, the work contributed improving existing tools for the study of circadian rhythms in mammals

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

Caracterização do ritmo de atividade/repouso do Mocó (Kerodon Rupestris) em fotoperíodo artificial

Kerodon rupestris (rock cavy, mocó) is an endemic caviidae of Brazilian northeast that inhabits rocky places in the semi arid region. The aim of this study was to characterize the activity/rest rhythm of the rock cavy under 12:12 h LD cycle and continuous light. In the first stage, seven animals were submitted to two light intensities (LD; 250:0 lux and 400:0 lux; 40 days each intensity). In the second stage four males were kept for 40 days in LD (470:<1 lux), for 18 days in LL 470 lux (LL470) and for 23 days in red dim light below 1 lux (LL<1). In the third stage three males were initially kept in LD 12:12 h (450:<1 lux) and after that in LL with gradual increase in light intensity each 21 days (<1 lux LL<1; 10 lux-LL10; 160 lux LL160; 450 lux LL450). In the fourth stage it was analyzed the motor activity of 16 animals in the first 10 days in LD. Motor activity was continuously recorded by passive infrared movement sensors connected to a computer and totaled in 5 min bins. The activity showed circadian and ultradian rhythms and activity peaks at phase transitions. The activity and the rest occurred in the light as well as in the dark phase, with activity mean greater in the light phase for most of the animals. The light intensity influenced the activity/rest rhythm in the first three stages and in the first stage the activity in 400 lux increased in four animals and decreases in two. In the second stage, the tau for 3 animals in LL470 was greater than 24 h; in LL<1 it was greater than 24 h for one and lower for two. In the third stage the tau decreased with the light intensity increase for animal 8. During the first days in the experimental room, the animals did not synchronize to the LD cycle with activity and rest occurring in both phases. The results indicate that the activity/rest rhythm of Kerodon rupestris can be affected by light intensity and that the synchronization to the LD cycle results from entrainment as well as masking probably as a consequence of the action of two or more oscillators with low coupling strength

O trato retino-hipotalâmico no mocó (Kerodon rupestris): Um estudo de traçado anterógrado com a subunidade b da toxina colérica

The light, besides the vision stimuli, controls other process completely independent of image formation, such as the synchronization of the organismic circadian rhythms to the enviromental light/dark cycle. In mammals, this adjust occurs through the retinohypothalamic tract, a direct retinal projection to the suprachiasmatic nucleus, considered to be the major circadian pacemaker. Early studies have identified only the suprachiasmatic nucleus as a retinal target in the hypothalamus. However, using more sensitive neuroanatomic tracers, other retinorecipient hypothalamic regions outside to suprachiasmatic nucleus were pointed in a great number of mammalian species. In this study, the retinohypothalamic tract was shown in the rock cavy (Kerodon rupestris), an endemic rodent of the semiarid region of the Brazilian Northeast, using unilateral intravitreal injections of cholera toxin subunit b as a neuronal tracer. The results reveal that in the rock cavy, besides the suprachiasmatic nucleus, several hypothalamic regions receive direct retinal projection, such as the ventrolateral preoptic nucleus, medial and lateral preoptic areas, the supraoptic nucleus and bordering areas, anterior, lateral and rectrochiasmatic hypothalamic areas, and the subparaventricular zone. The results are discussed by comparing with those of the literature, into a functional context

Caracaterização do núcleo pré-geniculado do sagüi (Callithrix jacchus) :projeção retiniana, neuroquímica e atividade celular (expressão de FOS)

In rodents, the suprachiasmatic nucleus (SCN) and the intergeniculate leaflet (IGL) are the main components of the circadian system. The SCN is considerate the site of an endogenous biological clock because can to generate rhythm and to synchronize to the environmental cues (zeitgebers) and IGL has been related as one of the main areas that modulate the action of SCN. Both receive projections of ganglion cells of retina and this projection to SCN is called retinohypothalamic tract (RHT). Moreover, the IGL is connected with SCN through of geniculohypothalamic tract (GHT). In primates (include humans) was not still demonstrated the presence of a homologous structure to the IGL. It is believed that the pregeniculate nucleus (PGN) can be the answer, but nothing it was still proven. Trying to answer that question, the objective of our study is to do a comparative analysis among PGN and IGL through of techniques immunohystochemicals, neural tracers and FOS expression after dark pulses. For this, we used as experimental model a primate of the new world, the common marmoset (Callithrix jacchus). Ours results may contribute to the elucidation of this lacuna in the circadian system once that the IGL is responsible for the transmission of nonphotic information to SCN and participate in the integration between photic and nonphotic stimulus to adjust the function of the SCN. In this way to find a same structure in primates represent an important achieve in the understanding of the biological rhythms in those animals

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

Sincronização dos ritmos de atividade motora e temperatura central à disponibilidade de glicose em ratos

Food is essential for the survival of all animals. Its temporal availability is an important enviromental cue for the behavioral and physiological organization throughout the 24 hours of day in different species. Rats and mice, for example, show increased locomotion in the hours before food availability when it is presented in a recurrent manner, a behavior named foodanticipatory activity. Several lines of evidence indicate that this anticipation is mediated by a circadian oscillator. In this work, based on the hypothesis that pre- or post-ingestive humoral signals are involved in the entrainment process, we tested whether the daily intake of glucose is sufficient to induce anticipatory activity in rats. The rhythms of motor activity and central temperature were recorded in animals undergoing 10 days of temporal glucose (solution at 50%) or chow restriction in light-dark (LD) and constant darkness (DD). Animals under temporal glucose restriction increase motor activity and and central temperature in the hours preceding glucose availability and such aticipation is extremely similar to that observed in animals under temporal chow restriction. Glucose ingestion is, therefore, a sufficient temporal cue to induce anticipation in rats. It is possible that the increase in plasma glucose after food ingestion constitutes one of the signals involved in the behavioral entrainment process to food availability

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

Influência de fotoperíodo artificial no comportamento de um primata neotropical diurno (Callithrix jacchus)

One of the main environmental cues for the adjustment of temporal organization of the animals is the light-dark cycle (LD), which undergoes changes in phase duration throughout the seasons. Photoperiod signaling by melatonin in mammals allows behavioral changes along the year, as in the activity-rest cycle, in mood states and in cognitive performance. The aim of this study was to investigate if common marmoset (Callithrix jacchus) exhibits behavioral changes under short and long photoperiods in a 24h cycle, assessing their individual behaviors, vocal repertoire, exploratory activity (EA), recognition memory (RM) and the circadian rhythm of locomotor activity (CRA). Eight adult marmosets were exposed to a light-dark cycle of 12:12; LD 08:16; LD 12:12 and LD 16:08, sequentially, for four weeks in each condition. Locomotor activity was recorded 24h/day by passive infrared motion detectors above the individual cages. A video camera system was programmed to record each animal, twice a week, on the first two light hours. From the videos, frequency of behaviors was registered as anxiety-like, grooming, alert, hanging position, staying in nest box and feeding using continuous focal animal sampling method. Simultaneously, the calls emitted in the experimental room were recorded by a single microphone centrally located and categorized as affiliative (whirr, chirp), contact (phee), long distance (loud shrill), agonistic (twitter) and alarm (tsik, seep, see). EA was assessed on the third hour after lights onset on the last week of each condition. In a first session, marmosets were exposed to one unfamiliar object during 15 min and 24h later, on the second session, a novel object was added to evaluate RM. Results showed that long days caused a decreased of amplitude and period variance of the CRA, but not short days. Short days decreased the total daily activity and active phase duration. On long days, active phase duration increased due to an advance of activity onset in relation to symmetric days. However, not all subjects started the activity earlier on long days. The activity offset was similar to symmetric days for the majority of marmosets. Results of EA showed that RM was not affected by short or long days, and that the marmosets exhibited a decreased in duration of EA on long days. Frequency and type of calls and frequency of anxiety-like behaviors, staying in nest box and grooming were lower on the first two light hours on long days. Considering the whole active phase of marmosets as we elucidate the results of vocalizations and behaviors, it is possible that these changes in the first two light hours are due to the shifting of temporal distribution of marmoset activities, since some animals did not advance the activity onset on long days. Consequently, the marmosets mean decreased because the sampling was not possible. In conclusion, marmosets synchronized the CRA to the tested photoperiods and as the phase angle varied a lot among marmosets it is suggested that they can use different strategies. Also, long days had an effect on activity-rest cycle and exploratory behaviors

Influência da auto seleção à luz no ritmo circadiano de atividade motora em Callithrix Jacchus

Marmosets, Callithrix jacchus, are strictly diurnal animals. The motor activity rhythmicity is generated by the circadian timing system and is modulated by environmental factors, mainly by photic stimuli that compose the light-dark cycle. Photic stimuli can reset the biological oscilators changing activity motor pattern, by a mechanism called entrainment. Otherwise, light can act directly on expressed rhythm, without act on the biological oscillators, promoting the masking. Thus, photic stimuli can synchronize the circadian activity rhythm (CAR) by two distinct mechanisms, acting isolated or at a combined way. Among the elements that can influence photic synchronization, the duration and time of photic exposure is pointed out. If in the natural environment the marmoset can choose places of different intensity illumination and is synchronized to light-dark cycle (LD), how the photic synchronization mechanism can be evaluated in laboratory by light self-selection? With objective to response this question, four adult male marmosets were studied at two conditions: with and without sleeping box. The animals were submitted to a LD cycle (12:12/ 350:2 lx) and constant light (LL: 350 lx) conditions in individual cages with an opaque sleeping box, that permitted the light self-selection. At the room, the temperature was 25.6 ºC (± 0.3 ºC) and humidity was 78.7 (± 5%). The motor activity was recorded at 5 min bins by infrared movement sensors installed at the top of the cages. The motor activity profile was distinct at the two conditions: without the sleeping box protection against light, the activity frequency was higher at CT 11-12 (ANOVA; F(3.23) = 62.27; p < 0.01). Also, the duration of the active phase (α) was prolonged of about 1 h (t test, p < 0.05) and the animals showed a significant delay on the activity onset and offset (t test, p < 0.05) and at the acrophase (confidence intervals of 5%) of CAR. In LL, the light continuous exposure prolonged the active phase and influenced the endogenous expression of the circadian activity rhythm period. From the result analysis, it is concluded that the light self-selection can modify several parameters of CAR in marmosets, allowing the study of the synchronization mechanism using the burrow model. Thus, without sleeping box there was a phase delay between the CAR and LD (entrainment) and an increase of activity near lights off (positive masking). Furthermore, in LL, the light continuous exposure modifies α and the endogenous expression of CAR. It is suggested that the light self-selection might be take into account at investigations that evaluate the biological rhythmicity in marmosets

Caracterização do perfil do ciclo sono-vigília em ratos sob dessincronização forçada

The circadian behavior associated with the 24 hours light-dark (LD) cycle (T24) is due to a circadian clock , which in mammals is located in the hypothalamic suprachiasmatic nucleus (SCN). Under experimental conditions in which rats are espoused to a symmetric LD 22h cycle (T22) the two SCN regions, ventrolateral (vl) and dorsomedial (dm), can be functionally isolated, suggesting that each region regulates distinct physiological and behavioral components. The vl region regulates the locomotor activity and slow wave sleep (SWS) rhythms, while the dm region assures the body temperature and paradoxical sleep (PS) rhythms regulation. This research aimed to deepen the knowledge on the functional properties of circadian rhythmicity, specifically about the internal desynchronization process, and its consequences to locomotor activity and body temperature rhythms as well as to the sleep-wake cycle pattern in rats. We applied infrared motion sensors, implanted body temperature sensors and a telemetry system to record electrocorticogram (ECoG) and electromyogram (EMG) in two rat groups. The control group under 24h period LD cycle (T24: 12hL-12hD) to the baseline record and the experimental group under 22h period LD cycle (T22: 11hL- 11hD), in which is known to occur the uncoupling process of the circadian locomotor activity rhythm where the animals show two distinct locomotor activity rhythms: one synchronized to the external LD cycle, and another expressed in free running course, with period greater than 24h. As a result of 22h cycles, characteristic locomotor activity moment appear, that are coincidence moments (T22C) and non coincidence moments (T22NC) which were the main focus or our study. Our results show an increase in locomotor activity, especially in coincidence moments, and the inversion of locomotor activity, body temperature, and sleep-wake cycle patterns in non coincidence moments. We can also observe the increase in SWS and decrease in PS, both in coincidence and non coincidence moments. Probably the increases in locomotor activity as a way to promote the coupling between circadian oscillators generate an increased homeostatic pressure and thus increase SWS, promoting the decreasing in PS

Efeito da suplementação com ácido linoléico conjugado e do treinamento em natação sobre a composição corporal e os parâmetros bioquímicos de ratos Wistar em crescimento

OBJETIVO: Avaliar os efeitos da suplementação com ácido linoléico conjugado, associada ao treinamento moderado em natação, sobre a composição corporal, o consumo e a eficiência alimentar, a glicemia, o perfil lipídico e o glicogênio muscular e hepático de ratos Wistar. MÉTODOS: Ratos Wistar (30 dias) foram divididos em: sedentário, sedentário suplementado, treinado e treinado suplementado. Permaneceram em gaiolas individuais com comida e água ad libitum, temperatura de 23ºC (com variação de1ºC) e ciclo claro-escuro de 12 horas, durante 8 semanas. A sessão de natação durou 1 hora e foi realizada três vezes/semana, bem como a suplementação com ácido linoléico conjugado a 2%. Após sacrifício, o plasma, os tecidos adiposos brancos e o marrom, o músculo gastrocnêmio e o fígado foram coletados e pesados. RESULTADOS: A suplementação per se não promoveu modificação na ingestão alimentar e na massa corporal dos animais. Houve aumento na glicemia de jejum (p<0,05), nas lipoproteínas de alta densidade (p<0,05), no colesterol total (p<0,05) e redução dos triacilgliceróis. A suplementação associada ao treinamento reduziu a massa corporal (p<0,05) e aumentou o peso relativo do tecido adiposo, do fígado e a glicemia de jejum. CONCLUSÃO: A suplementação com ácido linoléico conjugado associada à prática de exercício físico parece ter influência no balanço energético, mas, por outro lado, o aumento no peso do fígado indica que a ingestão deste ácido graxo pode ter efeitos indesejáveis, aumentando as chances de desenvolvimento do fígado gorduroso. Estes achados apontam perspectivas para novos estudos envolvendo análises histológicas do fígado, expressão gênica de enzimas chaves do metabolismo lipídico e de carboidratos, associados ou não a diferentes protocolos de treinamento físico.

O efeito de dois modelos de treinamento contínuo ou intervalado sobre metabolismo muscular e capacidade aeróbia de ratos obesos induzidos por dieta

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ação alelopática de saxitoxinas e microcistinas em cianobactérias e microalgas verdes em condições controladas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)