Relação entre padrão comportamental, estágios do ciclo de muda e atividade de enzimas digestivas proteolíticas em juvenis do camarão marinho Litopenaeus vannamei (Crustacea: Penaeidae)

Shrimp farming in Brazil is a consolidated activity, having brought economical and social gains to several states with the largest production concentrated in the northeast. This fact is also reflected in higher feed intake, necessitating a more efficient feed management. Currently, management techniques already foresee food loss due to molting. In this sense, studies relating shrimp s digestive physiology, molting physiology and behavioral response of shrimp feed can optimize the feed management. Thus, our study aimed to evaluate the behavioral response of the marine shrimp L. vannamei (Crustacea: Penaeidae) in accordance with the stages of moulting cycle and feeding schedules based on higher or lower activity of proteolytic digestive enzymes; also, to investigate the influence of feeding schedule on hepatosomatic index and non-specific and specific protease activity (trypsin). Experiments were carried out at the Laboratory of Shrimp Behavioral Studies at UFRN in partnership with the Laboratory of Enzimology UFPE. Juveniles of L. vannamei weighting 5.25 g (+ 0.25 g) were kept in aquaria at a density of 33 shrimp m -2. In the first experiment, shrimp were fed in the light phase or in the dark phase for 8 days; in the ninth day, the animals were observed for 15 minutes every hour during the 12 hours of each phase of the photoperiod. We recorded the frequency of inactivity, exploration, food intake, burrowing, swimming and crawling behavior. At the end of the 12th observation session, the shrimp were sacrified and classified by the method of setogenesis in the molt cycle stages A, B, C, D0, D1, D2 or D3. We found that the shrimp in A stage show high levels of inactivity. Moreover, the frequency of food intake was very low. The shrimp in D3 stage also had low food intake and high inactivity associated with elevated frequencies of burrowing. In the second experiment, shrimp were kept in physiological acclimation to experimental conditions for 28 days, distributed in 12 treatments in the light phase and 12 treatments in the dark phase. In the end, the animals were sacrified and dissected to assess non-specific and specific protease activity (trypsin) activity. In general, these parameters did not vary among animals fed in the light phase and those fed in the dark phase. However, significant differences were found in the activity of specific and nonspecific proteases in relation to food treatment. In the light phase, the major proteolytic activities converged to 10 hours after the start of the light phase, while the lowest activities converged to 6 hours after the beginning of this phase. In the dark phase, the highest enzyme activity converged to 12 hours after the onset of phase, while the lowest activities converged to 3 hours after the onset of phase. In the third experiment, we sought to evaluate the behavioral responses of shrimp in relation to dietary treatments based on higher or lower activity of proteolytic enzymes, considering the results of the second experiment. The behavioral categories observed were the same as the ones in the first experiment, with observations of 30 minutes (15min before and 15min after food supply). We found variation in behavioral responses as a function of the treatments, with greater intake of food in shrimp fed during the period of greatest activity of proteolytic enzymes, in the light phase. Thus we see that periodic events associated with the shrimp s physiology interfere in their behavioral responses, revealing situations that are more adjustable to the provision of food, and consequently optimizing feeding management

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

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

Influência da densidade de estocagem sobre o crescimento, ciclo de muda e o comportamento em juvenis do camarão marinho Litopenaeus vannamei

One of the factors that may interfere with the cultivation of Litopenaeus vannamei is the population density. This study aimed to assess the effect of density on growth, mortality, physical integrity and behavior of shrimp. The study was divided into two stages. At first, the shrimp were placed in tanks at densities of 50, 75 and 100 shrimp m-2. The animals were monitored in relation to the degree of proventricular filling, the stage of the molt cycle and physical integrity three times a week and in relation to the weight and length once a week. Mortality, growth and proventricular filling were not influenced by the density; frequency of records in specific stages of the molt cycle varied according to the density. The lower proportion of broken appendages and higher frequency of necrotic lesions occurred in lower density. The second stage of the research, conducted in aquaria, was divided into two parts. The first described social or feeding behavioral categories: slow displacement by contact, slow displacement by approximation, abrupt displacement by contact, abrupt displacement by approximation, reactivity, cannibalism, occupying the tray, get feed in the tray and get feed outside the tray. In the second part, these and other behavioral categories, described in the literature, were recorded in densities of 50, 75 and 100 shrimp m-2. Mortality was more frequent in higher density. The frequency of most behaviors mentioned above was very low, not differing between densities or being too low to determine differences between them. The behavioral profile of animals in different densities was, in general, very similar, with no difference in exploration, digging and cleaning between the densities. Even so, inactivity, feeding, crawling, burrowing, swimming, and proximity between animals were influenced by the density. These results suggest that some behaviors suffer greater interference from population density. However, the density may not have a broader influence on the animals when other factors, such as physico-chemical parameters of water and feed offering, are adequate

Efeitos do substrato e da densidade populacional sobre as atividades comportamentais e níveis de homócitos em relação à densidade em Litopenaeus vannamei (BOONE, 1931)

Carciniculture in Brazil occupies world-wide prominence due to shrimp culture, and the state of Rio Grande do Norte has presented the best results in the culture of the Litopenaeus vannamei in the last decade. This species has been shown to adapt easily to different environments and is between the five most cultivated penaeids of the world. The ponds are usually constructed in areas close to water courses and estuaries. Stock density and substrate ponds can pollute environment, causing losses in the growth and survival of the shrimps, being considered stress factors. Shrimps in inadequate densities and substrates can result reduced productivity of the farm; and favor diseases. So, it is important to verify how these variables influence the development of the animals in the culture farms. Our objective was to study the influence of the type of substrate and the stock density on the behavior and haemocyte count of the L. vannamei. Individually marked juvenile shrimps were kept in aquaria with 30 L of seawater and continuous aeration, in 12L-12D photoperiod. They were observed through Ad libitum and focal sampling instantaneous methods during thirty days, five times per week, six times per day (8:00 to 18:00) in windows of 15 minutes every two hours. The marking of carapace permitted quantifying molting and the feeding was supplied three times a day. Two experiments were carried out: the first one tested animals in the three different substrates (fine sand, smaller rocks-SPP and biggest rocks-SGR) with 33 shrimp/m2. In the second one, the animals were tested in three stock densities (26, 52 and 66 shrimp/m2) in fine sand substrate. At the end of experiment, biometry (first and second ones) and haemocyte count (second one) were made. The behavior of the L. vannamei seems to have been influenced by substrate and stocking density. In low granulometry of the substrate; the exploratory behavior became more frequent and inactivity of the shrimps was reduced. Burrowing was registered in sand substrate, specially in the initial period of the day. Cleaning was gradually higher along the day, presenting the biggest levels as the dark phase approached. The ingestion of feeding was more frequent in low density, and the animals were bigger and heavier at the end of the experiment. In the fine sand condition, the animals presented better growth, probably associated with the burrowing. The molting was equivalent in all types of substrate, but it was more frequent in high densities. Mortality of the shrimps was more frequent in high densities, and cannibalism and diseases were also registered in that condition. The clinical signals were similar to the ones of infectious mionecrosis (IMNV), generally associated with environment and physical stress. The haemocyte count was low for the hematologic standards of the penaeid, which we attributed for greater dilution of haemolymph in the postmolting phase. Smaller shrimps presented lower levels of haemocytes in relation to the bigger animals, count was also low in 26 shrimp/m2 density. The study demonstrates that stocking density and the granulometry of the substrate can affect the welfare, the health and the behavior of the L. vannamei. The sand substrate and low stocking density can be important tools in the management systems of shrimp production