Interaction between breeding habitat and elevation affects prevalence but not infection intensity of Batrachochytrium dendrobatidis in Brazilian anuran assemblages

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Haematologic and immunologic parameters of bullfrogs, Lithobates catesbeianus, fed probiotics

The effects of two probiotics (P(1) - Lactobacillus acidophilus, Bifidobacterium bifidum and Enterococcus faecium and P(2) - Bacillus subtilis) supplemented to commercial feed (40% crude protein) on the haematological and immunological parameters of the bullfrog Lithobates catesbeianus were studied. Two doses of each probiotic (5 and 10 g kg-1 of food) were added to the diets and fed to frogs, totalling five treatments over 112 days. Haematological analyses consisted of total and differential leucocyte counts, erythrocyte and thrombocyte counts, haematocrit, haemoglobin levels and RBC indices (mean corpuscular volume, mean corpuscular haemoglobin - and mean corpuscular haemoglobin concentration) and the immunological parameters included phagocytic capacity and phagocytic index of peritoneal phagocytes. The results showed that the probiotics did not significantly influence any of the haematological parameters measured. However, immunological assays showed that the probiotics had an immunostimulating effect. The greatest effects were seen with probiotic P(1) fed at a dose of 10 g kg-1 of diet and probiotic P(2) fed at 5 g kg-1 of diet.

Efeito do probiótico Bacillus subtilis no crescimento, sobrevivência e fisiologia de rãs-touro (Rana catesbeiana)

The probiotics are live microorganisms, in latency state, that benefices the development of the animals. The objective of this work was to evaluate the effect of the Bacillus subtilis in different doses about the growing, survival and immunological and hematological analyses in the Bullfrog (Rana catesbeiana) froglets. The test was developed in Experimental Frog farm of Agriculture Department, Pindamonhangaba, SP. Three doses of probiotics were tested (T1 - 2.5 g/Kg, T2 - 5 g/Kg and T3 - 10 g/Kg of food). The test was carried out with three simultaneous replicates, plus control group. The products were added to froglet's diet and mixed to the meal. The animals were previously feed with this diet for a period of 14 days and accompanied for about 42 days after metamorphose. Biometrics were performed every 7 days. It was evaluated the final weight, the survival, phagocytic capacity, phagocytic index and hematological analyses as hematocrit, hemoglobin level, erythrocyte number, absolute indices hematimetrycs (MCV and MCHC), differential counting of leucocytes and total counting of leucocytes and trombocytes. The results had shown that the doses of the probiotics had produced none effect on the final weight and survival. The immunological analyses had shown that the probiotics have presented immunostimulator effect, but haven't influenced the hematological parameters of the animals.

Influência da técnica de pressão expiratória positiva oscilante utilizando pressões expiratórias pré-determinadas na viscosidade e na transportabilidade do escarro em pacientes com bronquiectasia

OBJETIVO: Verificar a efetividade da técnica de pressão expiratória positiva oscilante (PEPO) utilizando pressões expiratórias pré-determinadas sobre a viscosidade e a transportabilidade do escarro em pacientes com bronquiectasia. MÉTODOS: Foram incluídos no estudo 15 pacientes estáveis com bronquiectasia (7 homens; média de idade = 53 ± 16 anos), submetidos a duas intervenções PEPO consecutivas, com 24 h de intervalo entre si, utilizando pressões expiratórias de 15 cmH2O (P15) e 25 cmH2O (P25). O protocolo consistiu de tosse voluntária; nova expectoração voluntária após 20 min, denominado tempo zero (T0); repouso de 10 min; e utilização da técnica em duas séries de 10 min (S1 e S2) de PEPO em P15 e P25, com intervalo de 10 min entre si. A viscosidade e transportabilidade do escarro foram avaliadas pela viscosimetria, velocidade relativa de transporte no palato de rã, deslocamento em máquina simuladora de tosse e ângulo de adesão. As amostras de escarro foram coletadas em T0, após S1 e após S2. Testes estatísticos específicos foram aplicados de acordo com a distribuição dos dados. RESULTADOS: Houve diminuição significante da viscosidade do escarro após S1 em P15 e após S2 em P25. Não houve diferenças significantes entre todas as amostras para a transportabilidade. CONCLUSÕES: Houve diminuição da viscosidade do escarro quando a PEPO foi realizada em P15 e P25, o que sugere que não seja necessário gerar alta pressão expiratória para obter o resultado desejado.

Encoding mechanisms based on fast oscillations in the retina of the cat and their dependencies on anesthesia

Processing in the visual system starts in the retina. Its complex network of cells with different properties enables for parallel encoding and transmission of visual information to the lateral geniculate nucleus (LGN) and to the cortex. In the retina, it has been shown that responses are often accompanied by fast synchronous oscillations (30 - 90 Hz) in a stimulus-dependent manner. Studies in the frog, rabbit, cat and monkey, have shown strong oscillatory responses to large stimuli which probably encode global stimulus properties, such as size and continuity (Neuenschwander and Singer, 1996; Ishikane et al., 2005). Moreover, simultaneous recordings from different levels in the visual system have demonstrated that the oscillatory patterning of retinal ganglion cell responses are transmitted to the cortex via the LGN (Castelo-Branco et al., 1998). Overall these results suggest that feedforward synchronous oscillations contribute to visual encoding. In the present study on the LGN of the anesthetized cat, we further investigate the role of retinal oscillations in visual processing by applying complex stimuli, such as natural visual scenes, light spots of varying size and contrast, and flickering checkerboards. This is a necessary step for understanding encoding mechanisms in more naturalistic conditions, as currently most data on retinal oscillations have been limited to simple, flashed and stationary stimuli. Correlation analysis of spiking responses confirmed previous results showing that oscillatory responses in the retina (observed here from the LGN responses) largely depend on the size and stationarity of the stimulus. For natural scenes (gray-level and binary movies) oscillations appeared only for brief moments probably when receptive fields were dominated by large continuous, flat-contrast surfaces. Moreover, oscillatory responses to a circle stimulus could be broken with an annular mask indicating that synchronization arises from relatively local interactions among populations of activated cells in the retina. A surprising finding in this study was that retinal oscillations are highly dependent on halothane anesthesia levels. In the absence of halothane, oscillatory activity vanished independent of the characteristics of the stimuli. The same results were obtained for isoflurane, which has similar pharmacological properties. These new and unexpected findings question whether feedfoward oscillations in the early visual system are simply due to an imbalance between excitation and inhibition in the retinal networks generated by the halogenated anesthetics. Further studies in awake behaving animals are necessary to extend these conclusions

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

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

Aspectos da fluência na narrativa oral de indivíduos com Transtorno do Espectro Alcoólico Fetal

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

Desempenho zootécnico e tabela de alimentação de girinos de rã-touro (Rana catesbeiana) criados no sistema anfigranja

Objetivou-se com a execução deste trabalho avaliar, em nível de campo, o desempenho zootécnico de girinos de rã-touro (Rana catesbeiana) criados em ranários comerciais que utilizam o sistema anfigranja, e simultaneamente estimar os valores de consumo diário de ração para compor uma tabela de referência para a alimentação dos animais. Os resultados encontrados nas unidades de observações monitoradas (aproximadamente 249 mil animais) atingiram os seguintes índices médios: a mortalidade (em %) variou de: 0,7 a 35,2 (média 5,9%); o ganho de peso (g. p/dia): de 0,03 a 0,22 (média 0,11 g/dia); e a conversão alimentar: de 0,92 a 2,75 : 1; (média 1,50 : 1). Comparados com dados preliminares, tais resultados indicam uma significativa otimização dos índices zootécnicos, demonstrando que houve um relativo ganho de produtividade com o aperfeiçoamento das técnicas de manejo implementadas. A redução do valor médio da conversão alimentar aparente foi provocada principalmente pelo adequado uso da tabela de referência, cujos valores foram ratificados durante o trabalho de monitoramento das unidades. Tal tabela possibilita o cálculo da quantidade de alimento a ser oferecido diariamente, baseado nos peso médio dos girinos e na estimativa do percentual de consumo, para temperatura da água entre 18 a 29 graus centígrados, (girinos de 0,1 a 26 gramas de peso médio).

Desempenho zootécnico e percentual de consumo de alimento de rã-touro (Rana catesbeiana) na fase de recria (pós-metamorfose) do sistema anfigranja

Objetivou-se com este trabalho avaliar (em nível de campo) o desempenho zootécnico da rã-touro, Rana catesbeiana (Shaw, 1882), criada em ranários comerciais que utilizam o Sistema Anfigranja, e , simultaneamente, estimar os valores de consumo diário de alimento para compor uma tabela de referência para o arraçoamento dos animais. As avaliações demonstraram haver valores de produtividade com grandes variações, de acordo com a capacidade do produtor em assimilar a tecnologia e principalmente manejar os animais de forma padronizada, para atingir o ponto de equilíbrio no fluxo de produção de seu plantel. Os resultados encontrados nas unidades de observação monitoradas (aproximadamente 250 mil animais) atingiram os seguintes índices médios: a mortalidade (em %) 0,5 a 39,5 (média geral 12,0); o ganho de peso (g./dia) 0,4 a 2,5 (média geral 1,2); e a conversão alimentar 0,9 a 2,5 : 1,0 (média geral 1,4 : 1,0). É apresentada uma tabela com os percentuais estimados de consumo de alimento, em função do peso médio dos animais, para o cálculo da quantidade de ração a ser oferecida diariamente.

Time interval from ovulation to extrusion in female bullfrog in different photoperiods

It was analyzed in this work the influence of photoperiod on time interval from ovulation induction period to extrusion of ovocits in female bullfrogs (Lithobates catesbeianus). It was used 54 females reared from metamorphosis to 9 months of age under three photoperiods: dark time (DL 0:24), 16 hours of daylight (DL 16:8) and 12 hours of daylight (DL 12:12). Ovulation was induced by intramuscular application of two doses of LHRHa with 12 hours of interval between the injections. After 10, 25, 28, 31, 34 and 37 hours from the first hormone injection, 10-gram samples (3,000 eggs) were extracted from each female at each time interval and fertilized. Egg hatching rate was checked in each sample 72 hours after fertilization. Analysis of variance showed a significant effect of extrusion delay and the interaction between photoperiod and this delay. Extrusion should be carried out 33, 24 and 26 hours after the first hormone dosage in females reared in environments without light, with 12 hours of daylight and with 16 hours of daylight, respectively, to obtain the maximum fertilization rate.