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

The evolution of jumping performance in anurans: morphological correlates and ecological implications

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

Morphological and physiological specialization for digging in amphisbaenians, an ancient lineage of fossorial vertebrates

Amphisbaenians are legless reptiles that differ significantly from other vertebrate lineages. Most species dig underground galleries of similar diameter to that of the animal. We studied the muscle physiology and morphological attributes of digging effort in the Brazilian amphisbaenid Leposternon microcephalum (Squamata; Amphisbaenia), which burrows by compressing soil against the upper wall of the tunnel by means of upward strokes of the head. The individuals tested (<72 g) exerted forces on the soil of up to 24 N. These forces were possible because the fibres of the longissimus dorsi, the main muscle associated with burrowing, are highly pennated, thus increasing effective muscle cross-sectional area. The muscle is characterized by a metabolic transition along its length: proximal, medial and distal fibres are fast contracting and moderately oxidative, but fibres closer to the head are richer in citrate synthase and more aerobic in nature. Distal fibres, then, might be active mainly at the final step of the compression stroke, which requires more power. For animals greater than a given diameter, the work required to compress soil increases exponentially with body diameter. Leposternon microcephalum, and probably some other highly specialized amphisbaenids, are most likely constrained to small diameters and can increase muscle mass and effective muscle cross-sectional area by increasing body length, not body diameter.

Osteology and Intraspecific Variation of Leptodactylus podicipinus (Anura: Leptodactylidae), with Comments on the Relationship between Osteology and Reproductive Modes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Non-local interactions and the dynamics of dispersal in immature insects

A simple mathematical model is developed to explain the appearance of oscillations in the dispersal of larvae from the food source in experimental populations of certain species of blowflies. The life history of the immature stage in these flies, and in a number of other insects, is a system with two populations, one of larvae dispersing on the soil and the other of larvae that burrow in the soil to pupate. The observed oscillations in the horizontal distribution of buried pupae at the end of the dispersal process are hypothesized to be a consequence of larval crowding at a given point in the pupation substrate. It is assumed that dispersing larvae are capable of perceiving variations in density of larvae buried at a given point in the substrate of pupation, and that pupal density may influence pupation of dispersing larvae. The assumed interaction between dispersing larvae and the larvae that are burrowing to pupate is modeled using the concept of non-local effects. Numerical solutions of integro-partial differential equations developed to model density-dependent immature dispersal demonstrate that variation in the parameter that governs the non-local interaction between dispersing and buried larvae induces oscillations in the final horizontal distribution of pupae. (C) 1997 Academic Press Limited.

Tafonomia de Pelecípodes da Formação Teresina (Permiano), Tiaraju, RS, Brasil

Taphonomic analysis of pelecypod concentration in the part of the Teresina Formation (Passa Dois Group), Tiaraju region, State of Rio Grande do Sul, indicates its origins as due to high energy events (storms). The fauna include shallow-burrowing suspensivorous species, associated with this byssate semi-infauna species. Several taphonomic characteristics indicate that the fossiliferous assemblage was subject to little selective processes during transportation. These are: predominance of disarticulated valves (although articulated valves are frequent) and perpendicular, oblique, concordant and nested arrangement of bioclasts in the sedimentary matrix. Absence of fragmentation, bioerosion and incrustation of the bioclasts, suggest fast burial of shells due to high sedimentation rate events. Diagenetic features, indicate that the fossils were later submitted to refossilization and mixing with other non-coeval bioclasts, resulting in some degree of time-averaging.

Controle do percevejo castanho (Scaptocoris castanea Perty, 1830 (Hemiptera: Cydnidae)) na cultura da soja (Glycine max (L.) Merrill)

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Estudo das alterações neuropatológicas e do comportamento em tarefas hipocampo-dependentes induzidas pela encefalite experimental aguda associada ao vírus piry

Este trabalho investiga as alterações neuropatológicas e do comportamento em tarefas hipocampo-dependentes induzidas pela encefalite experimental aguda associada ao vírus Piry. Três janelas temporais (3, 7 e 10 dias após a inoculação) foram avaliadas e dois ambientes testados de forma a avaliar se o enriquecimento ambiental influencia as alterações associadas à infecção. Camundongos fêmeas de dois meses de idade foram mantidos em ambientes empobrecido (IE) ou enriquecido (EE) durante seis meses e foram testados para as atividades de burrowing, de campo aberto e de discriminação olfatória. Após esse período os animais foram inoculados por via intranasal com 5μl de homogenado de cérebro normal (NBH) ou homogenado de cérebro infectado pelo vírus Piry (PY) e então reorganizados nos seguintes grupos: IENBH, IEPY, EENBH e EEPY, com sete animais cada. Três, sete e dez dias após a inoculação (dpi), os animais de cada janela temporal foram perfundidos com fixador aldeídico. Os encéfalos foram removidos, seccionados e as secções foram processadas para imunohistoquímica para anti-Piry e para anti-Iba-1 para marcação dos antígenos virais e de macrófagos/micróglias, respectivamente. Quantificações estereológicas foram feitas em cada camada celular de CA3 usando o método do fracionador óptico. Estimativas do fracionador óptico mostraram que não houve alteração da estimativa do número total de micróglias em CA3 nos grupos analisados, indicando que a infecção não alterou o número de células, mas a morfologia das micróglias, que se mostraram mais ativadas no grupo IEPY do que no grupo EEPY. Os resultados revelaram a presença de antígenos virais no bulbo olfatório, córtex piriforme, estriado e fimbria, ao longo da via olfatória. A atividade de burrowing no grupo IEPY diminuiu na primeira janela temporal e permaneceu baixa até a última janela, enquanto que no grupo EEPY não houve alteração neste teste. Na atividade de campo aberto, o grupo IEPY aumentou o tempo imóvel já na primeira janela e continuou aumentando até a última; reduziu o número de linhas cruzadas na segunda janela e permaneceu reduzido na última; e diminuiu o tempo na zona central na segunda e última janela. Já o grupo EEPY, aumentou o tempo imóvel e reduziu o número de linhas cruzadas na segunda janela. No teste de discriminação olfatória, o principal grupo afetado foi o grupo IEPY, que não discriminou os dois odores na última janela, enquanto que o grupo EEPY não teve alteração na discriminação.

Neurodegeneração crônica em modelo murino: ensaios comportamentais e neuropatológicos na doença prion em fêmeas adultas de camundongos albinos suíços

No presente trabalho implantou-se como modelo experimental para estudos de neurodegeneração crônica a doença prion induzida pelo agente ME7 em fêmea adulta no camundongo Suíço albino. As alterações comportamentais e neuropatológicas seguem de perto as previamente descritas para o camundongo C57BL/6j com duas exceções: 1) o septum ao invés do hipocampo é a região onde se detectou mais precocemente o maior número de microglias ativadas e astrócitos reativos e onde houve a maior redução de redes perineuronais nos estágios iniciais da doença; 2) Em relação ao C57BL/6j o curso temporal da doença é em média 4 semanas mais longo (26 semanas) e os sintomas iniciais começam a aparecer 4 semanas mais tarde (16 semanas) na variedade Suíça albina. Semelhante ao encontrado no C57BL/6j não se encontrou diferença nas estimativas do número de neurônios nos animais inoculados com o agente ME7 em relação aos inoculados com homogenado cerebral normal 15 e 1 8 semanas após a inoculação. A análise comparada do número de microglias ativadas astrócitos reativos e redes perineuronais empregando o fracionador óptico revelou diferenças significativas nos animais sacrificados na 15ª em relação aos sacrificados na 18ª semana pós-inoculação, com aumento do número das primeiras e redução do número das últimas na 18ª semana (teste T, bi-caudal p<0.05). A análise de cluster seguida da análise discriminante dos resultados dos testes comportamentais da variedade Suíça albina aplicada a cada quinzena ao longo do curso temporal da doença, revelou que a remoção de comida é a única variável discriminante para detecção de dois grupos distintos: um grupo menor (em torno de 40%, n=4), mais sensível, onde a doença cursa mais rápido e os animais atingem a fase terminal em 22 semanas, e outro maior (em torno de 60%, n=6), menos sensível, onde os animais atingem a fase terminal em 26 semanas. Os resultados são importantes para estudos comparativos de imunopatologia dentro da mesma e entre variedades de modelos murinos de neurodegeneração crônica na doença prion induzida pelo agente ME7.

Fácies e proveniência de depósitos costeiros da Formação Raizama: evidências do registro Ediacarano-cambriano na faixa Paraguai, região de Nobres, Mato Grosso

Rochas siliciclásticas da Formação Raizama, unidade basal do Grupo Alto Paraguai de idade ediacarana-cambriana (635 – 541 Ma), ocorrem distribuídas descontinuamente ao longo da margem sul do Cráton Amazônico e segmento norte da Faixa Paraguai, centro-oeste do Brasil, estado do Mato Grosso. Estas recobrem discordantemente os depósitos de plataforma carbonática do Grupo Araras, onde foram registrados evidências do evento glacial Marinoano (635 Ma). O Grupo Alto Paraguai representa os estágios finais da colisão entre os blocos Amazônico e Paranapanema que culminaram no fechamento do Oceano Clymene (540-520 Ma). A Formação Raizama com espessura de aproximadamente 570 m é constituída por pelitos, arenitos finos a grossos, e arenitos com cimento dolomítico previamente interpretados como depósitos flúvio-costeiros distribuídos nos membros inferior (270 m) e superior (300 m). O estudo faciológico e estratigráfico desta unidade na região de Nobres, Estado do Mato Grosso, foi focado principalmente na seção aflorante de 600 m no leito do rio Serragem, que inclui a Cachoeira da Serra do Tombador. Foram definidas 17 fácies sedimentares, agrupadas em cinco associações de fácies (AF) representativas de uma sucessão costeira progradante iniciando por depósitos de shoreface inferior, os quais recobrem em conformidade correlativa os depósitos de plataforma carbonática da Formação Serra do Quilombo (Grupo Araras). A AF1 consiste em arenitos com laminação plano-paralela e laminação truncada por onda (microhummocky), individualizada por camadas de pelito laminado interpretadas como depósitos de shoreface inferior. Destaca-se na AF1 a primeira ocorrência de níveis centimétricos bioturbados por Skolithos em depósitos neoproterozoicos – cambrianos na Faixa Paraguai. A AF2 é formada por arenitos com estratificação cruzada swaley e estratificação plano-paralela interpretada como depósitos de shoreface superior. A AF3 é composta por arenitos com estratificações cruzadas tangenciais e acanaladas com recobrimentos de siltito/arenito muito fino representativos de depósitos de canal e barras de submaré. A AF4 é caracterizada por arenitos com estratificações cruzadas tangencial e sigmoidal, laminação plano-paralela a cruzadas de baixo-ângulo, ritmito arenito muito fino/siltito com acamamento flaser e gretas de contração, organizados em ciclos métricos de raseamento ascendente de planície de maré. A AF5 é constituída por arenito com estratificação cruzada acanalada marcada por lags residuais na base da associação, arenito com estratificações plano-paralela e cruzada de baixo-ângulo, interpretados como depósitos fluviais distais de rios entrelaçados, parcialmente retrabalhados por ondas. Grãos detríticos de zircão foram obtidos da AF3 e datados pelo método U-Pb, sendo a idade de 1001±9 Ma interpretada como a idade de máxima deposição da Formação Raizama. Aliado a tal análise, as paleocorrentes NE-SE mostram que estes grãos teriam como áreas fontes principais a Faixa Sunsás, SW do Cráton Amazônico, não sendo descartada contribuições oriundas da parte NW desse Cráton. A idade mesoproterozóica obtida serviu principalmente para desvendar a proveniência da Formação Raizama, enquanto que as datações da base do Grupo Araras, em torno de 627-622 Ma, associada à presença inequívoca do icnogênero Skolithos, tornam esta unidade muito mais próxima do limite com o Cambriano Inferior. Traços fósseis do Proterozoico são caracterizados quase que exclusivamente por traços horizontais, sendo que bioturbações verticais praticamente são ausentes ao longo do Neoproterozoico. Esta inferência vem de encontro com a idade máxima de 541 Ma obtida para a Formação Diamantino, a qual recobre a unidade estudada. Os dados radiométricos aliados com as interpretações paleoambientais, que incluem o registro das primeiras atividades de organismos perfurantes na Faixa Paraguai, abrem perspectivas de entender com maiores detalhes a sequência de eventos que tipificam os estratos do limite Ediacarano-Cambriano do Brasil, ainda pouco conhecidos.