Expressão imuno-histoquímica de metaloproteína em carcinoma epidermóide de lábio inferior e língua

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Expressão imuno-histoquímica da e-caderina e do CD44v6 em carcinoma epidermóide de lábio inferior e língua

The objective of this study was perform, by the streptoavidin-biotin technique, an immunohistochemical analysis of the E-cadherin and CD44v6 in 15 lower lip squamous cell carcinomas and 15 of tongue, with varied histologic gradation of malignidade, in order to establish a possible relation between the expression these proteins and the anatomical localization of the lesions, metastasis, as well as with the Bryne`s histolologic grading of malignancy system. It was not observed significant statistical association between the localization of the lesions and the malignancy score, however, had a significant correlation between the histologic parameters of malignancy gradation and the total score of malignancy, being that the parameter degree of keratinization presented the highest correlation (r = 0,844). Taking in consideration the anatomical localization of the lesions, it was not significant difference between the profile of expression and the amount of immunopositive cells for Ecaderina and CD44v6. To the metastasis variable, also it was not observed significant difference between the profile of expression and the amount of immunopositive cells for evaluated proteins. However, it was observed a statistical significant difference in relation to the scores of malignancy, being that the low score presented the highest values for the profile of expression and the amount of immunopositive cells for the E-caderina and the CD44v6. It was observed a statistically significant and negative correlation between the expression profile, the amount of E-cadherin and CD44v6 immunopositive cells and the total score of malignancy. Therefore, based in the results of this study, it was concluded that the expression of the immunohistochemical markers E-caderina and CD44v6 did not constitute histological indicator of aggressiveness for the patients with lower lip and tongue squamous cell carcinomas

Expressão imuno-histoquímica da e-caderina e da ß-catenina em carcinoma epidermóide oral com e sem metástase nodal

The adhesion molecules E-cadherin and β-catenin have been studied as possible markers to distinguish carcinomas with and without metastatic potential. The objective of this research was to study the imunohistochemistry expression of the E-cadherin and β-catenin in oral squamous cell carcinoma (OSCC), aiming to contribute for the better understanding of the biological behavior of this lesion. The sample consisted of 30 cases of OSCC, being 15 of tongue and 15 of lower lip. The profile and intensity of labeling and semi quantitative analysis of the percentage of immunopositive tumoral cells in membrane for E-cadherin and β-catenin had been related with the anatomical localization of the lesion, the presence or not of nodal metastasis and the histological grade of malignancy in the invasive front area of the tumor. It was registered the presence or not of cytoplasmic and nuclear labeling of the β-catenin. The results had been submitted to the statistical analysis, being used the Mann-Whitney Test, the Fisher Test and the Spearman Correlation Coefficient (α=0, 05). The results showed that the expression in membrane for E-cadherin and β-catenin was, predominantly, the heterogeneous profile in the lower lip and tongue carcinomas, as well as in the cases with and without nodal metastasis. It was not observed significant statistical difference between expression profile and amount of immunopositive cells for E-cadherin, β-catenin and the anatomical localization of the lesion and for the presence or not of nodal metastasis. However, there was significant difference of the reduced expression of these proteins with the high score of malignancy. It was verified that the expression of the β-catenin in cytoplasm was present in 22 (73.33%) of the 30 analyzed cases, and 6 cases (20%) showed nuclear expression. The statistical analysis detected significant association between the expression of the β-catenin in the cytoplasm with the histological grade of malignancy, being this molecule more frequently present in the cytoplasm in the cases of high score of malignancy. It was concluded that the reduced immunoexpression of these proteins in membrane can be related with the lowest cellular differentiation, as well as with the pattern of invasion in nests and isolated cells, demonstrated in the cases of OSCC with high histological grade of malignancy

Expressão imuno-histoquímica das integrinas a2ß1, a3ß1 e a5ß1 em adenoma pleomórfico de glândula salivar menor e maior e carcinoma adenóide cístico

Pleomorphic adenoma and adenoid cystic carcinoma (ACC) consist benign and malignant neoplasm from salivary gland, respectively. These neoplasms share some characteristics, such as cellular origin and considerable production of extracellular matrix, however, with distinct biological behavior. The aim of the present study was to compare the expression of D2E1, D3E1 e D5E1 integrins in pleomorphic adenoma from minor and major salivary glands and ACCs. Furthermore, it was investigated possible differences in the expression of these integrins according to histological subtypes of ACC. Fourteen cases of pleomorphic adenoma from major salivary gland, fourteen cases from minor salivary gland and ten cases of ACC were selected. It was taken into consideration the presence or absence, localization and intensity of integrin immunoexpression. The cases of pleomorphic adenoma were grouped in order to compare the expression between the distinct neoplasms. It was observed a highly significant difference (p<0,0001) in relation to D2E1 integrin between the neoplasms since pleomorphic adenoma showed a pronounced immunostaining. It was not possible to perform statistical tests considering the D2E1 integrin expression; nevertheless, it could be observed a tendency of higher staining in pleomorphic adenoma. For comparative reasons the cases ACCs were divided in two groups: solid and tubular/cribriform. It was not detected significant differences in regard to D2E1 integrin; and statistical analysis could not be realized in relation to D3E1 and D5E integrin expression. However, it was also verified a tendency of absence or reduced expression in the solid subtype. It can be concluded that the reduced D2E1 integrin expression observed in CACs may be related to a lesser degree of cell differentiation in this neoplasm and the reduced D5E1 integrin expression can be associated with aggressive biological behavior. Moreover, the absence and/or reduced expression of the studied integrins in solid ACC suggests a role in pathogenesis and more aggressive biological behavior of this histological subtype

Perfil imuno-histoquímico do infiltrado inflamatório no front de invasão em carcicomas epidermóides de língua e lábio inferior

The progression of the oral squamous cells carcinomas (OSCCs) seems to suffer influence from related factors to the host, as local and systemic immunologic response, which are essential to the antineoplasic defenses. The purpose of this study was evaluate the local immunity in 30 tongue and 20 lower lip SCC by immunohistochemistry method, utilizing antibodies anti-CD3, CD4, -CD8, -CD25 e -ζ(zeta), which immunoexpressions were compared considering the anatomical localization, the intensity of the inflammatory infiltrate into the front of invasion and metastases. The CD4/CD8+ ratio was calculated for each case and associate with the mentioned variable, being the intensity of the inflammatory infiltrated also compared with the anatomical localization and metastase and for this the cases had been grouped in two categories: (n = 10) absent/scarce inflammatory infiltrate; and (n = 40) moderate/intense infiltrate. Fisher´s exact test was performed (α= 0.05) and it was not observed any significant correlation between these groups with anatomical sites and metastases. With regard to the immunoexpression, the CD3+, CD4+, CD8+ and CD25+ cells count was higher in the lower lip SCCs while the anti-ζimmunomarcation was more evident in the non metastatic cases. Through the statistical analyses, it was verified that the CD3 exhibited positive-significant correlation with the inflammatory infiltrate (p = 0.023). Furthermore, antibodies against CD8 and CD25 cells were also significantly correlated with the inflammatory infiltrate (p = 0.002 and 0.030, respectively) and with the anatomical site (p = 0.004 and p = 0.004) mainly in the lower lip SCCs. CD4/CD8 ratio did not show significant association with metastase nor with anatomical localization. We conclude that the inflammatory infiltrated of the Bryne s (1998) system did not constitute an indicator of aggressiveness in the tongue and lower lip SCCs analyzed and that clinical behavior of the SCCs studied was related in part to the immunohistochemical profile of infiltrated the inflammatory present in tumoral invasion front

Diferenças funcionais das regiões hipocampais na formação da memória do tipo o quê", "quando" e "onde" em ratos

Episodic memory refers to the recollection of what, where and when a specific event occurred. Hippocampus is a key structure in this type of memory. Computational models suggest that the dentate gyrus (DG) and the CA3 hippocampal subregions are involved in pattern separation and the rapid acquisition of episodic memories, while CA1 is involved in memory consolidation. However there are few studies with animal models that access simultaneously the aspects ‗what-where-when . Recently, an object recognition episodic-like memory task in rodents was proposed. This task consists of two sample trials and a test phase. In sample trial one, the rat is exposed to four copies of an object. In sample trial two, one hour later, the rat is exposed to four copies of a different object. In the test phase, 1 h later, two copies of each of the objects previously used are presented. One copy of the object used in sample trial one is located in a different place, and therefore it is expected to be the most explored object.However, the short retention delay of the task narrows its applications. This study verifies if this task can be evoked after 24h and whether the pharmacological inactivation of the DG/CA3 and CA1 subregions could differentially impair the acquisition of the task described. Validation of the task with a longer interval (24h) was accomplished (animals showed spatiotemporal object discrimination and scopolamine (1 mg/kg, ip) injected pos-training impaired performance). Afterwards, the GABA agonist muscimol, (0,250 μg/μl; volume = 0,5 μl) or saline were injected in the hippocampal subregions fifteen minutes before training. Pre-training inactivation of the DG/CA3 subregions impaired the spatial discrimination of the objects (‗where ), while the temporal discrimination (‗when ) was preserved. Rats treated with muscimol in the CA1 subregion explored all the objects equally well, irrespective of place or presentation time. Our results corroborate the computational models that postulate a role for DG/CA3 in spatial pattern separation, and a role for CA1 in the consolidation process of different mnemonic episodes

Expressão de fos após pulso de escuro no núcleo pré- geniculado do tálamo do sagui (Callithrix jacchus)

The pregeniculate nucleus (PGN) of the primate s thalamus is an agglomerate neuronal having a cap shaped located dorsomedially to the main relay visual information to the cerebral cortex, the dorsal lateral geniculate nucleus (GLD). Several cytoarchitectonic, neurochemical and retinal projections studies have pointed PGN as a structure homologous to intergeniculate leaflet (IGL) of rodents. The IGL receives retinal terminals and appears to be involved in the integration of photic and non-photic information relaying them, through geniculo-hypothalamic tract (TGH), to the main circadian oscillator in mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus. Thus, the IGL participates in the control of the biological rhythm by modulating the activity of the SCN. Pharmacological and IGL injury studies conclude that it is critical in the processing of non-photic information which is transmitted to the SCN. Other studies have found that especially neurons immunoreactive to neuropeptide Y (NPY) respond to this type of stimulation, determined by its colocation with the FOS protein. Has not been determined if the PGN responds, expressing the FOS protein, to the non-photic stimulus nor the neurochemical nature of these cells. Thus, we apply a dark pulse in the specifics circadian phases and analyze the pattern of expression of FOS protein in PGN of the marmoset (Callithrix jacchus). We found that in all animals analyzed the FOS expression was higher in the experimental than in the control group. There was a higher expression of FOS when the dark pulse was applied during the subjective day between the groups. Still, a subregion of the PGN, known by immunoreactive to NPY, had a greater number of FOS-positive cells in relation to his other just close dorsal region. Our data corroborate the theory that the PGN and IGL are homologous structures that were anatomically modified during the evolutionary process, but kept its main neurochemical and functional characteristics. However, injury and hodological studies are still needed for a more accurate conclusion

A Zona Incerta no sagüi (Callithrix jacchus): Análise Citoarquitetônica, Neuroquímica e Projeção Retiniana

The retinal projections in mammals usually reach, classically, three major functional systems: the primary visual system, the accessory optic system, and the circadian timing system. But the retinal projections also reach areas classically considered non-visual, one of which groups the neurons of the zona incerta (ZI), target this study. The primary visual system includes thalamic lateral geniculate complex is formed by the dorsal lateral geniculate nucleus, intergeniculate leaflet and the ventral lateral geniculate nucleus and other Components. The accessory optic system is composed of the small nuclei: nuclei terminal dorsal, lateral, medial and the interstitial nucleus of the superior posterior fasciculus. These nuclei are involved in visuo-motor activities. The circadian timing system is comprised of the suprachiasmatic nucleus of the hypothalamus, that act as master circadian pacemaker, entraining pathways and efferents pathways to the efectors, and the intergeniculate leaflet, that seems to act as a modulator of the pacemaker. The retinal projections too reach classically considered non-visual areas, including the zona incerta. This region is localized in the ventral thalamus and has been implicated in various functional properties including nociceptive and somatosensory processing, motor response, sociosexual behaviour, feeding and drinking, in symptoms of neurodegenerative diseases, arousal and attention. It also displays connection with several areas of central nervous system. The aim of this study was characterize the retinal projection in the zona incerta of Callithrix jacchus (sagüi), a primate of the New World through the anterograde axonal transport of the cholera toxin subunit b and analyze the citoarchicteture using Nissl and NeuN, and neurochemical substances such as serotonin, GABA, VIP, VP, GFAP and binding-calcium proteins. The zona incerta showed a different division of the literature in citoarquitetura, both by means of Nissl as neurochemical by NeuN, with a subdivision ventrolateral and dorsomedial. The neurochemical to the other substances corroborate with this subdivision. The GFAP was almost completely negative for the zona incerta, result non evidenced in previous studies yet. The 16 retinal projection in sagüi, unlike other primates and rodents, reached the caudal portion only. This work helps to make further studies are conducted based on this subdivision and the localization of the neurochemical substances associated with possible behaviors that the zona incerta is involved

Distribuição diária das atividades comportamentais dos camarões marinhos Farfantepenaeus subtilis (Pérez Farfante, 1967) e Litopenaeus schmitti (Burkenroad, 1938) em condições laboratoriais

Marine aquaculture has developed in the last decades all over the world, especially related to shrimp management. In Brazil, the introduction of the species Litopenaeus vannamei has contributed to the success of the activity, even if there are problems associated with the management of a exotic species, such as new diseases and ecological pressure on native species. It has been emphasized the need of research for developing new methodology that will allow native species management, being the most important Farfantepenaeus subtilis and Litopenaeus schmitti. Most knowledge obtained from research with those species has generally used a technical approach and mainly focused feeding process. There are no specific behavioral data on their activity pattern which should be of great importance for the use of native species on commercial culture farms. So, it was our objective to study and compare the daily distribution of behavioral activities of the marine shrimp species Litopenaeus schmitti and Farfantepenaeus subtilis. Forty animals of each species, 5 individuals per aquarium, were maintained in aquaria containing 200L of sea water under continuous aeration and filtration. They were marked individually and were observed by the instantaneous focal time sampling, along 10 continuous days, in 6 daily 15 min observation windows, every two hour. In each window, behaviors and location position of the animals in the aquarium were registered at 1 min intervals. Food was offered 3 times a day, representing 10% of each aquarium biomass. Aquaria were maintained in artificial photoperiod, 12hour light/l2 hour dark, 4 aquaria in light cycle equivalent to the environmental one (light from 06:00 to 17:59 h and dark from 18:00 to 05:59 h) and the other 4 in the reverse light cycle (light from 18:00 to 05:59 h and dark from 06:00 to 17:59 h) to allow sequential behavioral observation in both phases of the 24 hour cycle. There was a clear distinction between the distribution of behavioral activities of F. subtilis and L. schmitti in the two phases. The activity pattern of Farfantepenaeus subtilis demonstrates that species has prominently night habits and a burying pattern during the light cycle. Exploration, inactivity and swimming were the most common activities. The behavioral pattern of Litopenaeus schmitti indicates that species is active along both phases of the light cycle, and the most evident behaviors were exploration, inactivity and swimming

Neurofisiologia hipocampal durante sessões de sono intercaladas por exploração espacial: Reverberação do passado ou predição do futuro?

The ability to predict future rewards or threats is crucial for survival. Recent studies have addressed future event prediction by the hippocampus. Hippocampal neurons exhibit robust selectivity for spatial location. Thus, the activity of hippocampal neurons represents a cognitive map of space during navigation as well as during planning and recall. Spatial selectivity allows the hippocampus to be involved in the formation of spatial and episodic memories, including the sequential ordering of events. On the other hand, the discovery of reverberatory activity in multiple forebrain areas during slow wave and REM sleep underscored the role of sleep on the consolidation of recently acquired memory traces. To this date, there are no studies addressing whether neuronal activity in the hippocampus during sleep can predict regular environmental shifts. The aim of the present study was to investigate the activity of neuronal populations in the hippocampus during sleep sessions intercalated by spatial exploration periods, in which the location of reward changed in a predictable way. To this end, we performed the chronic implantation of 32-channel multielectrode arrays in the CA1 regions of the hippocampus in three male rats of the Wistar strain. In order to activate different neuronal subgroups at each cycle of the task, we exposed the animals to four spatial exploration sessions in a 4-arm elevated maze in which reward was delivered in a single arm per session. Reward location changed regularly at every session in a clockwise manner, traversing all the arms at the end of the daily recordings. Animals were recorded from 2-12 consecutive days. During spatial exploration of the 4-arm elevated maze, 67,5% of the recorded neurons showed firing rate differences across the maze arms. Furthermore, an average of 42% of the neurons showed increased correlation (R>0.3) between neuronal pairs in each arm. This allowed us to sort representative neuronal subgroups for each maze arm, and to analyze the activity of these subgroups across sleep sessions. We found that neuronal subgroups sorted by firing rate differences during spatial exploration sustained these differences across sleep sessions. This was not the case with neuronal subgroups sorted according to synchrony (correlation). In addition, the correlation levels between sleep sessions and waking patterns sampled in each arm were larger for the entire population of neurons than for the rate or synchrony subgroups. Neuronal activity during sleep of the entire neuronal population or subgroups did not show different correlations among the four arm mazes. On the other hand, we verified that neuronal activity during pre-exploration sleep sessions was significantly more similar to the activity patterns of the target arm than neuronal activity during pre-exploration sleep sessions. In other words, neuronal activity during sleep that precedes the task reflects more strongly the location of reward than neuronal activity during sleep that follows the task. Our results suggest that neuronal activity during sleep can predict regular environmental changes

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

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

A influência do status social no forrageio de itens camuflados e avermelhados em sagüis (Callithrix jacchus) cativos

Seeing colors can be advantageous, because they are an important signal for providing information about the environment, such as the location of food. However, not every animals sees these chromatic signals in the same way. In primates, the group of Plathyrrhini has polymorphic sex-linked vision with males always dichromats and dichromats or trichromats females. Studies indicate that trichromats during foraging would benefit by seeing better than dichromats ripe fruits against the green foliage background. On the other hand, dichromats appear to distinguish camouflaged insects better than trichromats. The marmoset (Callihtrix jacchus) is a neotropical primate species that have color vision polymorphism. This species establishes family groups with highly reproductive bias, with breeding females often having preferential access to food. This work aims to study whether the social context influences the foraging ability of camouflaged and red items in groups of C. jacchus. Four groups of captive marmosets were presented to four food tasks, involving difficult, easy, reddish and camouflaged food targets. Foods were presented in a concentrated and dispersed manner, to check whether there was monopolization of the resources by the dominant subjects and if this would affect the ability of individuals to find the food targets. Success was measured by latency to food acquisition and number of targets consumed. Males and females differed in their foraging success for camouflaged and reddish items, although this difference has not appeared in all situations and experimental conditions. In general males were more successful for detecting camouflaged items while females succeeded more in identifying reddish items. There were no differences in foraging success between individuals of different social status, however, there were differences in the success of consumption of food items for different situations when food was concentrate compared with dispersed food. Taken as a role, there was a greater difficulty in detecting food items when they were presented in concentrated arrangement, which is supposed to be related to a higher difficulty to approach and stay near the food. Although it appears that there was no direct competition seems to have group's indirect influence on the detection of food items and foraging success of individuals, affecting mainly those items more difficult to detect

Otimização no uso de martelos e bigornas para quebrar sementes por macacos prego (Cebus flavius e C. libidinosus) no Bioma Caatinga

Recently, capuchin monkeys (Cebus spp.) inhabitants of dry environments and with restriction of fleshy fruits, have been the subject of several studies regarding the use of instruments. During behaviour of using stones to crack open nuts there is evidence of selection of more effective hammers, as well as selection of anvils related to reducing the risk of predation. The aim of this study was to determine whether two groups of capuchin monkeys (C.flavius and and C.libidinosus) inhabitants of the Caatinga of Rio Grande do Norte make choice of hammers and anvils. The record of weight and location of stones indicated active choices of with what (choice of hammers) and where (selection of anvils) to crack open encapsulated seeds. The choice of hammers to break nuts depended on the type and degree of ripeness seed. Thus, smaller seeds were smashed with lighter hammers and larger seeds with heavier hammers. Still, C. flavius was the only species that presented a refinement in the choice of hammers that depended on the ripeness of seeds. For both species of capuchin monkeys studied, the nut-crack sites were not spread in accordance with the spatial distribution of seed-producing species, suggesting that the capuchin monkeys promote active choice of anvils. Thus, in environments with more escape routes through the trees, the nut-crack sites were found further apart than in regions that had less chance of escape through the trees. Also, there was a difference in the spacing of the anvils to depend on the type of seed: sites used to crack larger and more caloric seeds were found farther apart than the sites used to crack smaller and less caloric seeds, suggesting a pattern of avoiding direct competition. We conclude that the capuchin monkeys maximize energy savings and reduced risk of predation and the costs of food competition during the behaviour of using stones to crack open nuts

Delimitação dos grupamentos serotoninérgicos/núcleos da rafe do mocó (kerodon rupestris): citoarquitetura e imunoistoquímica para serotonina

Serotonin or 5-hydroxytryptamine (5-HT) is a substance found in many tissues of the body, including as a neurotransmitter in the nervous system, in which may exert varied post-synaptic actions. Inside the neuro-axis, the location of 5-HT neurons is almost restricted to the raphe nuclei of the brainstem, such that 5-HT-immunoreactivity can be considered a marker of the raphe nuclei. The raphe nuclei are located in the brainstem, at or near the midline. The serotonergic groups were originally alphanumerically classified as B1 to B9 towards caudorrostral in rats and can be divided into upper and lower groups. In this study the distribution of serotonergic neurons was studied using immunohistochemistry in the brain of the rock cavy (Kerodon rupestris), a species of rodent endemic to Northeastern Brazil. The cytoarchitectonic location of serotonergic neurons was established in series of adjacent coronal and sagittal sections stained by the Nissl method and immunohistochemistry for 5-HT. Thus, we defined the raphe rostral linear, caudal linear, dorsal, median, and paramedian pontine raphe nuclei, and B9 cluster, constituting the rostral group, and the interpositus, magnus, obscure and palidus, constituting the caudal part of the group, comparable to which has been described for other mammalian species

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