The ultrastructural evaluation of digestive cells in the hepatopancreas of the Amazon River prawn, Macrobrachium amazonicum

The Amazon River prawn Macrobrachium amazonicum is endemic to tropical South America and is being intensively exploited by artisanal fisheries in Brazil. Limited information is available about the nutritional requirements of M. amazonicum, although the production of this species is now technically feasible. The digestive process in this species is still unclear and investigation into the digestive cells of its hepatopancreatic epithelium is required. Thus, the hepatopancreas from 15 specimens were fixed in Karnovsky Solution and processed for Transmission Electron Microscopy. Our results indicate that E cells are located at the distal portion of the hepatopancreatic tubule and are involved in mitotic activity. The cylindrical R cells are sparse and are only found in the proximal portion of the hepatopancreatic tubule. According to its ultrastructural characteristics, this cell is involved in pinocytosis. M cells are generally found near the R cells. The F cells are scattered throughout the length of the hepatopancreatic tubules, and B cells are observed mainly in the proximal and middle regions. F cells and B cells are likely related to, respectively, the synthesis of enzymes and the intracelular digestion. R- and M cells are probably related to material storage. Thus, these findings provide basic information on the cell types that perform protein digestion in M. amazonicum, and will be useful in further nutritional research. The identification and characterization of digestive cells is an important step towards understanding the digestive mechanisms.

Influência de diferentes substratos sobre a composição, a densidade e as interações tróficas do zooplâncton em lagoas de policultivo de camarão-de-água-doce Macrobrachium amazonicum e tilápia-do-nilo Oreochromis niloticus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ontogenetic Variation in Ammonia Excretion during the Early Life Stages of the Amazon River Prawn, Macrobrachium amazonicum

Dry mass (DM) and total ammonia-N (TAN) excretion were determined in embryos, larvae (ZI-ZIX, Z = zoea ), and postlarvae (PL) at 1, 7, and 14 d after metamorphosis (PL1, PL7, and PL14) of Macrobrachium amazonicum. Animals in postmolt-intermolt (A-C) stages were sorted according to their developmental stages, and placed into incubation chambers (similar to 30 mL) for 2 h to quantify TAN excretion. After this period, analyses were carried out using Koroleff`s method for TAN determination. Individual TAN excretion generally increased throughout ontogenetic development and varied from 0.0090 +/- 0.0039 mu g TAN/individual/h in embryo to 1.041 +/- 0.249 mu g TAN/individual/h in PL14. There was no significant difference between embryo-ZIV and ZV-ZIX (P > 0.05), whereas PL1, PL7, and PL14 differed (P < 0.05) from each other. Higher increments in individual ammonia-N excretion were observed between ZIV-ZV, PL1-PL7, and PL7-PL14. Mass-specific excretion rates presented two groups, embryo-ZII (P > 0.05) and ZIII-PL14 (P > 0.05). The lowest value was found in embryo (0.17 +/- 0.07 mu g TAN/mg DM/h) and the maximum values in ZV and PL1 (0.65 +/- 0.25 and 0.64 +/- 0.27 mu g TAN/mg DM/h, respectively). Results indicate that metabolic rate is proportional to the body mass in M. amazonicum, during early life stages. Variations in ammonia excretion during this phase may be associated mainly with body size. Data obtained in the present study may be useful in developing and optimizing rearing techniques of M. amazonicum, such as the proportions between biofilter and rearing tank size, and stocking density in culture tanks or in transport bags.

Taxonomic revision of doubtful Brazilian freshwater shrimp species of genus Macrobrachium (Decapoda, Palaemonidae)

The freshwater prawns of the genus Macrobrachium Spence Bate, 1868 are widely distributed in rivers of tropical and subtropical regions and represent an interesting group with controversial taxonomy. The morphological characters traditionally used to separate species have shown a high intraspecific variation. Doubts about the status of M. birai Lobao, Melo & Fernandes, 1986, M. holthuisi Genofre & Lobao, 1978 and M. petronioi Melo, Lobao & Fernandes, 1986 have been arisen due to the high resemblance of the former two species with M. olfersi (Wiegmann, 1836), and the latter one with M. potiuna (Muller, 1880). Therefore, we performed a detailed morphological analysis of these species, including new characters not usually used in the species recognition. The present results here with molecular data lead us to conclude that M. birai and M. holthuisi are junior synonyms of M. olfersi, and M. petronioi is a junior synonym of M. potiuna. Considering these synonymies, 17 valid species are now reported for the Brazilian territory.

Densidade de estocagem em camarões da espécie Macrobrachium rosenbergii (De Man, 1879) na fase juvenil

Among the species of freshwater shrimp being cultivated, Macrobrachium rosenbergii stands out. Knowledge about the behavior of this species and the influence of certain factors on its development can help optimize management practices and minimize the likely impacts shrimp farming has on the environment and the animals themselves. The objective of this study was to characterize the species' behavior during early stages of development under different stocking densities over a 24-hour cycle. Ten day old postlarvae were transferred from the Jundiaí School of Agriculture (EAJ - Escola Agrícola de Jundiaí) in Macaíba (RN), Brazil to the Shrimp Behavior Laboratory (LSPR - Laboratório de Estudos do Comportamento do Camarão) at the Federal University of Rio Grande do Norte (UFRN), where they were weighed and measured. Eight aquaria with constant temperature, aeration and filtration, and subjected to a12 h light/12 h dark cycle were used for each experiment. Each aquarium also contained two shelters made of bricks and the water quality was monitored weekly. Behavioral observations were made at two densities: 25 individuals/m-² and 40 individuals/m-². The methods for recording behaviors were: behavioral sampling – enter and leave the shelter, exploring on the substrate, exploring in the water column, move away, attack, pursuit and cannibalism; scan sampling - inactivity, feeding, exploration, digging, swimming, cleaning and staying in the shelter. Observations were made during a 15 minute period/per aquarium at a frequency of 4 times daily, for 4 days/week, and over 4 weeks. Food was provided 2 times/day for each aquarium population, immediately before the 1st and 3rd observation periods. Our results demonstrate that at high density, there is an increased frequency of agonistic behavior; during the light phase, there is a greater frequency of behaviors that result in less exposure (inactivity, cleaning and staying in the shelter); during the dark phase, there is an increased frequency of behaviors that result in greater exposure (feeding, exploration, swimming and digging); at times of feed offer, there is an increased frequency of leaving the shelter, moving away, pursuit, feeding, exploration and swimming. At low density, the animals showed a lower frequency of agonistic behaviors, greater weight gain and higher growth rates, which indicates that this is a more favorable growing environment for cultivation and when applied, can generate better living conditions, favor survival rates and increase management success

Influência do tipo de abrigo e da frequência de oferta alimentar no comportamento do camarão Macrobrachium rosenbergii (De Man 1879)

Macrobrachium rosenbergii is a freshwater prawn which presents agonistic behavior and heterogeneous growth. It is known that captive conditions can intensify agonism causing injuries and decreased survival, generating a condition of poor welfare. Based on this, we aim to investigate the behavior of M. rosenbergii in the juvenile phase according to different types of shelter and frequencies of feed offer, emphasizing their agonistic behavior. For this, juveniles were observed in the laboratory in three steps. At step I we characterized the behavioral profile; prawns were kept in eight aquariums (27 prawns/m2 ), identified and observed four times along both phases of 24 h light cycle. At step II (2 experiments), we evaluated the use of shelters (brick or polyethylene rolls) and their influence on agonism by the animals. For classification of animals in dominance rank, the method used was David's Score. At step III (3 experiments), we evaluated different frequencies of feed offer on the behavior of individuals, in particular agonism. Results showed that juveniles do not present a pattern activity/inactivity between the phases of the light cycle. We identified a dominance hierarchy among individuals taking advantage of access to food by the dominant, which showed greater weight gain although the frequency of intake did not differ between individuals. The type of shelter influenced the behavior of animals. Brick shelter generated a higher frequency of permanence and a reduction in the frequency of agonistic interactions. The distribution of food more frequently throughout the day, decreased the motivation of animals for food, as well as to fight. Prawns fed four times showed lower frequency of feed intake and agonistic interactions. Thus, we conclude the shelters which reduce animal’s detection by coespecifics and offer the food four times along the day reduce agonistic behavior. This result causes na improvement in life quality of the prawns and also in its quality as final product.

Desempenho zootécnico dos camarões: Litopenaeus vannamei e Macrobrachium rosenbergii em tanques rede e aquicultura familiar na comunidade de Bebida-Velha, RN

The study aims to evaluate the performance and identify lesions of agonistic interactions of Macrobrachium rosenbergii in different densities cages (chapter 1); compare the growth performance of shrimp M. rosenbergii and Litopenaus vannamei cages (chatper 2); in addition the study aimed at identifying the knowledge and practices of a traditional community of farmers Bebida-Velha, in the city of Pureza / RN, practicing rural family and aquaculture, as well as tracing the community socioeconomic profile and evaluate the management that these farmers use in shrimp farming Macrobrachium rosenbergii (chapter 3). To perform the experiments (chapter 1 and 2) shrimp species L. vannamei and M. rosenbergii were weighed on a digital scale and transferred to cultures in cages. The cages had 1m³, with 5mm mesh between us and were closed on top with screen to avoid predators (birds, insects). The animals remained in adaptation for fifteen days before the start of data collection (each experiment). Both experiments lasted seventy days, totaling eighty-five days cultivation. During the cultures, animals were fed pelleted feed for shrimp at 10% of their biomass with 35% crude protein, offered in feed trays twice a day during the hours (7:00 am and 14:00 pm). The remnants in the tray were removed after 2h of permanence to calculate intake. To determine the performance parameters, some samples were taken every 10 days. The results of both experiments were analyzed using the STATISTIC 7.0 (2004). In Chapter 1 experiment were applied5 treatments were applied with 5 replications each: D5 - 5 animals/m2 ; D10 - 10 animals/m2 ; D15 - 15 animals/m2 ; D20 - 20 animals/m2 ; D25 - 25 animals/m2 . To this, were distributed 25 cages randomly in two masonry nurseries. The end of the experiment the lesions were verified and quantified. The chapter 2 experiment began when the shrimp reached the same age (eighty-four days) and was used the density of 25 shrimp / m2 with 4 repetitions. For the realization of chapter 3 were applied semi-structured interviews by direct approach of Bebida Velha of settlers practicing aquaculture activity, tilapia producers and community shrimp. Data were tabulated and analyzed according to the responses obtained by the participants. Therefore, the amount of damage increased with increasing stocking density. The density of 10 freshwater shrimp/m² showed the best conditions for a better performance in cages. It can be concluded that in cages, the cultivation of species of fresh water shrimp M. rosenbergii had better zootechnical indexes than the cultivation of marine shrimp species L. 9 vannamei. It was possible to verify that the activities of which the interviewees practice guarantee a good quality of life and income for them. We find that respondents have traditional and local knowledge, and also may be interested in the cultivation of the species M. rosenbergii.

Influence of stocking densities in the nursery phase on the growth of Macrobrachium amazonicum reared in net pens

The Amazon river prawn (Macrobrachium amazonicum) is a native species with great potential for aquaculture, based on promising results obtained from experimental culture trials in Brazil. The influence of different stocking densities on the development of prawns reared in cages in the nursery phase as well as on their growth when transferred to grow-out net pens at lower densities was evaluated. In the nursery phase, juveniles I (0.157 +/- 0.014 g, 47 days old) were stocked in 0.5 m(-2) cages at densities of 400, 800 and 1200 m(-2). After 71 days, prawns were transferred to grow-out net pens of 2.0 m(-2), at a density of 20 juveniles II m(-2). The treatments were determined by the mean weights registered for the prawns (118 days old) previously stocked at 400, 800 and 1200 juveniles I m(-2) in the nursery phase: 0.94 +/- 0.07 g (T1), 0.61 +/- 0.04 g (T2) and 0.48 +/- 0.07 g (T3), respectively. In the nursery phase, mean survival was above 96%, whereas mean weights were significantly higher (P<0.05) for the density of 400 prawns m(-2). The highest biomass (276.7 g) and productivity (1152 juveniles II m(-2)) were registered at the density of 1200 prawns m(-2), differing significantly (P<0.05) from the lower densities. One month after the transfer of the animals to the net pens, there was recovery in the specific growth rate (SGR) of prawns in all treatments which was significantly higher (P<0.05) in T3 (4.01 +/- 0.36% day(-1)) and T2 (3.60 +/- 0.18% day(-1)). The feed conversion efficiency (FCE) in the first month after the transfer was also significantly higher (P<0.05) in T3 (78.2 +/- 19.1%) when compared to T1 (39.8 +/- 9.5%). These results suggest the occurrence of a compensatory growth in M. amazonicum after transferring them to lower densities, which can point out high densities for nursery cages as a viable practice. After 277 days of grow-out phase in net pens in the cold season, survival, mean weight and biomass did not differ significantly among the treatments, indicating the viability of using net pens in stocking prawns during autumn and winter, since the minimum temperature does not drop below 17 degrees C. Influence of stocking density during the nursery phase on the grow-out of prawns was not observed. The population structure in prawns reared in net-pens was similar to that observed in earthen ponds. (C) 2012 Elsevier B.V. All rights reserved.

The Predation of Artemia Nauplii by the Larvae of the Amazon River Prawn, Macrobrachium amazonicum (Heller, 1862), is Affected by Prey Density, Time of Day, and Ontogenetic Development

The present study analyzed the effects of prey density, the time of day, and ontogenetic development on the predation of Artemia nauplii by the larvae of the Amazon river prawn, Macrobrachium amazonicum, as well as possible synergy among these factors. Larvae were raised in 120-L tanks with biological filter systems, and fed on recently hatched Artemia nauplii, using two feeding management protocols: (a) fed once per day at 2000 h (high density HD) and (b) half of the ration provided at 2000 h, complemented at 0800 h the following day by a replacement of the nauplii consumed up to a maximum of the full ration (low density with replacement LDWR). Each treatment consisted of six replicates. The consumption of nauplii was estimated prior to the feeding times. Consumption varied according to time of day, ontogenetic development, and feeding protocol. The larvae ingested more nauplii during the daytime at most developmental stages. Ingestion rates were similar during the day under both treatments, but at night the higher density of prey in the HD treatment caused a higher encounter rate and increased ingestion of nauplii by the larvae. Among the performance indicators only survival was greater in HD in comparison with LDWR; productivity and dry weight were similar. The results indicate a circadian trophic rhythm in M. amazonicum, with the encounter rate being an important mechanism for the capture of prey during the night. A second mechanism probably the visual system aids the perception of prey during the daytime. Based on these results, we suggest that feeding captive Amazon river prawn larvae only once a day would be appropriate and economically beneficial. Further work is necessary to determine the most effective time that this single feed should be applied.

Efeito da intensidade luminosa no desenvolvimento larval do camarão-da-amazônia, Macrobrachium amazonicum

The effects of four light intensities (0; 2.8 +/- 0.9; 5,5 +/- 1,8 e 7,8 +/- 2,5 mu mol s(-1) m(-2), about 136.5 +/- 87.5; 273 +/- 43.8 e 390 +/- 125 lux, respectively) on survival, productivity, weight gain and larval development of Macrobrachium amazonicum were investigated. Four treatments with three replicate tanks were evaluated. Newly hatched larvae were held in black tanks (80.2 +/- 0.6 larvae L(-1)) filled with 50-L-brackish water (salinity of 10), in a recirculating system. Tanks were covered with shadow cloth allowing 35% and 70% light, respectively, to reach light intensities of 2.8 +/- 0.9 and 5.5 +/- 1.8 mu mol s(-1) m(-2) at the water surface. Complete absence of light (0 mu mol s(-1) m(-2)) was obtained covering the tanks with opaque black plastic, and full-light condition used no covering (7.8 +/- 2.5 mu mol s(-1) m(-2)). Observations showed that the survival rate was not affected by light intensity. Productivity and weight gain were higher under 7.8 +/- 2.5 mu mol s(-1) m(-2) light intensity than under 0 and 2.8 +/- 0.9 1 mu mol s(-1) m(-2) intensities (P<0.05). The larval development index was similar among the treatments under the different light intensities. However, from stage VII this index was increased slightly in the treatment under 7.8 +/- 2.5 mu mol s(-1) m(-2) light intensity. In conclusion, light intensity affects larval development of M. amazonicum. Values as high as 7.8 mu mol s(-1) m(-2) (about 390 lux) improve the larval performance by enhancing development, productivity and weight gain compared to lower values.

Food intake of Macrobrachium rosenbergii during larval development

This work investigates the acceptance of different food types and sizes by Macrobrachium rosenbergii during each larval stage. Food intake of dry and wet formulated diets of four different size classes (250-425, 425-710, 710-1000 and 1000-1190 mum), as well as Artemia nauplii, was determined. Larvae of each zoeal stage were stocked in beakers and fed ad libitum. After 30-45 min, the digestive tract of each larva was observed under a stereomicroscope. Acceptance was evaluated by food intake frequency (FFI). There was no significant interaction (P<0.05) between inert diet size and FFI for each larval stage. Therefore, food intake during larval development is independent of food particle size. The ingestion of Artemia nauplii, was significantly higher by larvae between stages II and VI. Between stages VII and XI, FFI for Artemia nauplii and wet diet was similar, while the FFI of the dry diet was similar to live food between stages IX and XI. The wet diet was ingested by more than 50% of the larvae only from stage VII onwards, while the dry diet from stage VIII onwards. These results indicate that larvae could be fed Artemia nauplii only until stage VI. Diet supplementation should start from stage VII onwards, using food particles varying from 250 to 1190 mum. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Ingestion rates of Artemia nauplii for different larval stages of Macrobrachium rosenbergii

The functional response between ingestion rate and food concentration was determined for each larval stage of Macrobrachium rosenbergii. Artemia franciscana nauplii were supplied at 2,4, 6, 8, 10 and 12 per milliliter. The nauplii were counted by sight using a Pasteur pipette and transferred to Petri dishes containing 40 ml of brackish water (12 parts per thousand) lying on the top of black plastic. One larva at each stage was individually placed into each Petri dish containing different food density. After 24 h, each larva was removed from the Petri dish and the leftover nauplii were counted. The amount consumed was determined by the difference between the initial and final number of nauplii. Ingestion rate (I) increased as food density (P) increased and was defined by the model I=I-m(1-e(-kP)). The results suggest four levels of ingestion during larval development. The first level includes stages II, III and IV, with average maximum consumption of about 40 nauplii/day; the second level includes stages V and VI, with consumption of approximately 55 nauplii/day; the third level includes stages VII and VIII, with consumption of 80-100 nauplii/day. The fourth level includes stages IX, X and XI, in which the high values for maximum ingestion (Im) exceed the load capacity of the medium. The low values for constant k (that may correspond to the adaptability of the food to prey characteristics, such as, size, mobility, etc.) obtained for stages IX, X and XI indicated that Artemia is not an adequate prey and there is necessity of a supplementary diet. The best relationship between predator and prey seemed to occur during stage IV Results obtained in the present work may subsidize future researches and serve as a guideline for practical considerations of feeding rates. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Comportamento alimentar do camarão de agua doce, Macrobrachium rosenbergii (De Man) (Crustacea, Palaemonidae) durante a fase larval: análise qualitativa

Feeding behavior of the larvae's prawn, Macrobrachium rosenbergii de Man, 1879, in each stage of development was studied face alArtemia Linnaeus, 1758 nauplii, dry and wet diets. Groups of ten larvae in each stage were placed in 500 ml beakers at 29ºC in filtered brackish water. After the fasting period of acclimatization, the three types of food were offered separately to the larvae and the feeding behavior was observed during 30 min. Feeding perception, capture, grasp and ingestion of food were used to determine feeding pattern. Stage I lawac do not eat. The feeding behavior of larvae was similar on the stages II - XI. The perception in distance does not occur in most individuais; the feeding process generally started with the physrcal contact of food with the animal. Food is captured by thoracic appendages (maxillipeds until stage III, and maxillipeds + pereopods in stages IV - XI) and particles are grasped and handled by maxillipeds. Inopposition at the capture, the ingestion is a seletive process. Selective responses decrease in later stages and larvae become omnivorous in greater degree.

Uso de bandejas de alimentação na fase de engorda do camarão-da-Amazônia Macrobrachium amazonicum (HELLER, 1862)

The aim of this experiment was to evaluate the hypothesis that the M. amazonicum prawns accept feeding in trays. Six ponds were stocked with 10 juveniles II (1.2 +/- 0.7 g) per m(2) in 02/14/2003. An entirely randomized experimental design with 2 treatments (feeding in trays or feeding to the throw) and 3 replicates was used. Prawn average weight, survival, productivity, and physical and chemical variables of the water were compared between treatments using "t" test of Student. Average weight, survival and productivity were, respectively, 7.2 +/- 0.3 g, 46.8 +/- 7.8% and 335 +/- 45 kg/ha in feeding tray treatment and 6.8 +/- 0.1 g, 85.8 +/- 5.2% and 586 +/- 42 kg/ha in feed to the throw treatment. Survival and productivity differed significantly (p<0.5), while that average weight didn't differ significantly. M. amazonicum needs to use further number of trays per area, possibly due to territorial and aggressive behavior.

Sistema de recirculação e rotina de manejo para larvicultura de camarões de água doce Macrobrachium rosenbergii em pequena escala

A recirculating system and a matching management schedule for small-scale freshwater prawn larviculture were described. The system comprised a 140 L larval culture tank in line with a 43 L biofilter filled with 24 L of calcareous substrate. Both the tank and biofilter were made of black colored fiberglass in a conical-cylindrical shape. The turnover rate of the water through the system was 24 times a day; water was pumped by airlift. Results of ten larvicultures showed that the system maintained temperature, dissolved oxygen, pH, salinity, ammonium and nitrite stable and suitable to Macrobrachium rosenbergii larvae. Survival and productivity varied from 60.5 to 72.4% and 37 to 72 post-larvae/L, respectively; both were compatible with results of commercial hatcheries. Therefore, this system may be very useful for research purposes or adapted for small-scale post-larvae production.