Abstracts
Probably as a function of their wide geographical distribution, the different population of Macrobrachium amazonicum shrimp may present distinct physiological, biochemical, reproductive, behavioral, and ecological patterns. These differences are so accentuated that the existence of allopatric speciation has been suggested, although initial studies indicate that the genetic variability of populations happen at an intraspecific level. Among the biological responses described for M. amazonicum populations, those regarding osmoregulation and metabolism play a key role for being related to the occupation of diverse habitats. To this effect, we investigated osmoregulation through the role of free amino acids in cell volume control and metabolism, through oxygen consumption in larvae (zoeae I, II, V and IX) and/or post-larvae of a M. amazonicum population from Amazon, kept in aquaculture fish hatcheries in the state of São Paulo. The results add information regarding the existence of distinct physiological responses among M. amazonicum populations and suggest that possible adjustments to metabolism and to the use of free amino acids as osmolytes of the regulation of the larvae and post-larvae cell volume depend on the appearance of structures responsible for hemolymph osmoregulation like, for example, the gills. In this respect, we verified that zoeae I do not alter their metabolism due to the exposition to fresh or brackish water, but they reduce intracellular concentration of free amino acids when exposed to fresh water, what may suggest the inexistence or inefficient performance of the structures responsible for volume regulation and hemolymph composition. On the other hand, in zoeae II and V exposed to fresh and brackish water, metabolism alterations were not followed by changes in free amino acids concentration. Thus it is possible, as the structures responsible for osmoregulation and ionic regulation become functional, that the role of free amino acids gets diminished and oxygen consumption elevated, probably due to greater energy expenditure with the active transportation of salts through epithelial membranes. Osmotic challenges also seem to alter throughout development, given that in zoeae II oxygen consumption is elevated on brackish water of 18, but in zoeae V it happens in fresh water. After M. amazonicum metamorphosis, free amino acids begin to play an important role as intracellular osmolytes, because we verified an increase of up to 40% in post-larvae exposed to brackish water of 18. The main free amino acids involved in cell volume regulation of ontogenetic stages evaluated were the non essential ones: glutamic acid, glycine, alanine, arginine, and proline. Interestingly, larvae from estuarine population studied here survived until the zoeae V stage in fresh water, but in some populations far from the sea, zoeae die right after eclosion in fresh water or they do not reach zoeae III stage. In addition, given that in favorable conditions caridean shrimp larvae shorten their development, we may infer that the cultivation environment, in which larvae developed in the present work, was appropriate, because almost all zoeae VIII kept on brackish water underwent metamorphosis directly to post-larvae and did not go through zoeae IX stage.
Macrobrachium ; osmorregulation; Crustacea; metabolism; physiology
Provavelmente como função da sua ampla distribuição geográfica, as diferentes populações do camarão Macrobrachium amazonicum podem apresentar distintos padrões fisiológicos, bioquímicos, reprodutivos, comportamentais e ecológicos. Essas diferenças são tão acentuadas que tem sido sugerido a existência de especiação alopátrica embora estudos iniciais indiquem que a variabilidade genética das populações ocorre ao nível intraespecífico. Dentre as respostas biológicas descritas para as populações de M. amazonicum, aquelas relacionadas à osmorregulação e metabolismo têm papel central por estarem relacionadas à ocupação dos diversos habitats. Nesse sentido, investigou-se a osmorregulação, por meio do papel dos aminoácidos livres no controle do volume celular e o metabolismo, por meio do consumo de oxigênio, em larvas (zoeas I, II, V e IX) e/ou pós-larvas de uma população de M. amazonicum oriunda da Amazônia e mantida em viveiros de aquicultura no estado de São Paulo. Os resultados adicionam informações a respeito da existência de respostas fisiológicas distintas entre as populações de M. amazonicum e sugerem que possíveis ajustes no metabolismo e no uso de aminoácidos livres como osmólitos da regulação do volume celular das larvas e pós-larvas dependem do surgimento de estruturas responsáveis pela osmorregulação da hemolinfa como, por exemplo, as brânquias. Nesse sentido, verificou-se que as zoeas I não alteram seu metabolismo em função da exposição à água doce ou salobra, mas reduzem a concentração intracelular de aminoácidos livres quando expostas à água doce, o que pode sugerir a inexistência ou um desempenho ineficiente das estruturas responsáveis pela regulação do volume e composição da hemolinfa. Por outro lado, nas zoeas II e V expostas à água doce ou salobra alterações no metabolismo não foram acompanhadas por mudanças na concentração dos aminoácidos livres. Assim é possível que à medida que estruturas responsáveis pela osmo e ionorregulação tornam-se funcionais, o papel dos aminoácidos livres se torne reduzido e o consumo de oxigênio elevado, provavelmente em função do maior gasto energético com o transporte ativo de sais através das membranas epiteliais. Os desafios osmóticos também parecem se alterar ao longo do desenvolvimento visto que em zoeas II o consumo de oxigênio é elevado em água salobra de 18 mas em zoeas V essa resposta ocorre em água doce. Após a metamorfose de M. amazonicum, os aminoácidos livres passam a ter papel importante como osmólitos intracelulares, pois se verificou um aumento de até 40% nas pós-larvas expostas à água salobra de 18. Os principais aminoácidos livres envolvidos na regulação do volume celular dos estágios ontogenéticos avaliados foram os não essenciais ácido glutâmico, glicina, alanina, arginina e prolina. Interessantemente, as larvas da população estuarina aqui estudada sobrevivem até o estágio de zoea V em água doce mas em algumas populações distantes do mar as zoeas morrem logo após a eclosão em água doce ou não chegam ao estágio de zoea III. Adicionalmente, visto que em condições favoráveis as larvas de camarões carídeos abreviam o seu desenvolvimento pode ser inferido que o meio de cultivo em que as larvas se desenvolveram no presente trabalho foi adequado, pois quase todas as zoeas VIII mantidas em água salobra sofreram diretamente a metamorfose para pós-larvas e não passaram pelo estágio de zoeas IX.
Macrobrachium ; osmorregulação; Crustacea; metabolismo; fisiologia
1 Introduction
The M. amazonicum shrimp has a wide geographical distribution that
goes from Caribbean and Atlantic coasts of South America to northern Argentina and
Paraguay and the eastern slopes of Andes in Ecuador, Bolivia and Peru to the
Atlantic coasts of northeastern Brazil (Maciel and
Valenti, 2009Maciel, CR. and Valenti, WC., 2009. Biology, fisheries, and
aquaculture of the Amazon River Prawn Macrobrachium amazonicum:
a review. Nauplius, vol. 17, p. 61-79.). There is a geographical separation and, in consequence, a
genetic isolation among M. amazonicum populations of the northern
region (also including the Atlantic and Caribbean coasts and bays of Amazon and
Orinoco) and southern region (La Plata System) of Brazil. This wide geographical
distribution has as consequence the existence of distinct physiological,
reproductive, behavioral, and ecological patterns among many populations. In
relation to reproduction, Urzúa and Anger
(2011)Urzúa, A. and Anger, K., 2011. Larval biomass and chemical
composition at hatching in two geographically isolated clades of the shrimp
Macrobrachium amazonicum: intra- or interspecific
variation?Invertebrate Reproduction & Development, vol. 55, no. 4, p.
236-246. http://dx.doi.org/10.1080/07924259.2011.576155.
http://dx.doi.org/10.1080/07924259.2011....
verified differences in biomass and chemical composition of larvae
of M. amazonicum populations from Pantanal (state of Mato Grosso,
Brazil) and Belém (state of Pará, Brazil). In relation to physiology, it is
observed, among the different M. amazonicum populations, a curious
pattern of brackish water dependence, in which there are either populations that
complete their life cycle in fresh water (Zanders
and Rodriguez, 1992Zanders, IP. and Rodriguez, JM., 1992. Effect of temperature and
salinity stress on osmoionic regulation in adult and on oxygen consumption in
larvae and adult of . Macrobrachium amazonicum (Decapoda,
Palaemonidae)Comparative Biochemistry and Physiology. A. Comparative Physiology,
vol. 101, no. 3, p. 505-509.
http://dx.doi.org/10.1016/0300-9629(92)90502-H.
http://dx.doi.org/10.1016/0300-9629(92)9...
; Charmantier and
Anger, 2011Charmantier, G. and Anger, K., 2011. Ontogeny of osmoregulatory
patterns in the South American shrimp . Macrobrachium
amazonicum: loss of hypo-regulation in a land-locked population
indicates phylogenetic separation from estuarine ancestorsJournal of
Experimental Marine Biology and Ecology, vol. 396, no. 2, p. 89-98.
http://dx.doi.org/10.1016/j.jembe.2010.10.013.
http://dx.doi.org/10.1016/j.jembe.2010.1...
) and those in which larvae die when kept in this salinity
(Augusto et al., 2007aAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
). Charmantier and Anger (2011)Charmantier, G. and Anger, K., 2011. Ontogeny of osmoregulatory
patterns in the South American shrimp . Macrobrachium
amazonicum: loss of hypo-regulation in a land-locked population
indicates phylogenetic separation from estuarine ancestorsJournal of
Experimental Marine Biology and Ecology, vol. 396, no. 2, p. 89-98.
http://dx.doi.org/10.1016/j.jembe.2010.10.013.
http://dx.doi.org/10.1016/j.jembe.2010.1...
compared two
M. amazonicum populations hydrologically and genetically
separated and verified distinct osmoregulation and tolerance to salinity patterns
among them. Urzúa and Anger (2011)Urzúa, A. and Anger, K., 2011. Larval biomass and chemical
composition at hatching in two geographically isolated clades of the shrimp
Macrobrachium amazonicum: intra- or interspecific
variation?Invertebrate Reproduction & Development, vol. 55, no. 4, p.
236-246. http://dx.doi.org/10.1080/07924259.2011.576155.
http://dx.doi.org/10.1080/07924259.2011....
suggested
that the existing biological diversity among M. amazonicum
populations may happen due to the limited genetic exchange among them, a case of
allopatric speciation. Although the studies about the genetics of different
populations are still incipient, Vergamini et al.
(2011)Vergamini, FG., Pileggi, LG. and Mantelatto, FL., 2011. Genetic
variability of the Amazon River prawn (Decapoda, Caridea, Palaemonidae).
Macrobrachium amazonicumContributions to Zoology
(Amsterdam, Netherlands), vol. 80, no. 1, p. 67-83. verified that the genetic variability of coast and countryside
M. amazonicum populations happen at an intraespecific
level.
The wide distribution of M. amazonicum followed by the diversity of
biological responses makes this specie a very interesting object to be studied.
Among physiological mechanisms which can be investigated, osmoregulation and
metabolism have a notable importance for being related to the distribution of the
species in different environments. In this sense, osmotic and ionic regulation in
crustaceans is composed of two processes, extracellular anisosmotic regulation,
responsible for the maintenance of the osmolality and performed by the action
epithelial enzymes like Na+/K+- ATPase, V-ATPase,
HCO3-ATPase, carbonic anhydrase (Freire et al., 2008Freire, CA., Onken, H. and McNamara, JC., 2008. A structure-function
analysis of ion transport in crustacean gills and excretory organs. Comparative
Biochemistry and Physiology. Part A, Molecular & Integrative Physiology,
vol. 151, no. 3, p. 272-304. http://dx.doi.org/10.1016/j.cbpa.2007.05.008.
PMid:17604200
http://dx.doi.org/10.1016/j.cbpa.2007.05...
; Garçon
et al., 2013Garçon, DP., Lucena, MN., Pinto, MR., Fontes, CFL., McNamara, JC.
and Leone, FA., 2013. Synergistic stimulation by potassium and ammonium of
K(+)-phosphatase activity in gill microsomes from the crab Callinectes ornatus
acclimated to low salinity: novel property of a primordial pump. Archives of
Biochemistry and Biophysics, vol. 530, no. 2, p. 55-63.
http://dx.doi.org/10.1016/j.abb.2012.12.006. PMid:23262318
http://dx.doi.org/10.1016/j.abb.2012.12....
) and intracellular isosmotic regulation, responsible for the
maintenance of intracellular media through the adjustment of the concentration
intracellular osmolytes, mainly free amino acids (Péqueux, 1995Péqueux, A., 1995. Osmotic regulation in crustaceans. Journal of
Crustacean Biology, vol. 15, no. 1, p. 1-60.
http://dx.doi.org/10.2307/1549010.
http://dx.doi.org/10.2307/1549010...
; McNamara et al.,
2004Mcnamara, JC., Rosa, JC., Greene, LJ. and Augusto, A., 2004. Free
amino acid pools as effectors of osmotic adjustment in different tissues of the
freshwater shrimp . Macrobrachium olfersii (Crustacea,
Decapoda) during long-term salinity acclimationMarine and Freshwater Behaviour
and Physiology, vol. 37, no. 3, p. 193-208.
http://dx.doi.org/10.1080/10236240400006208.
http://dx.doi.org/10.1080/10236240400006...
; Augusto et al., 2007aAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
,
bAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007b.
Adaptative shifts in osmoregulatory strategy and the invasion of fresh water by
brachyuran crabs: evidence from . Dilocarcinus pagei
(Trichodactylidae)Journal of Experimental Zoology. Part A, Comparative
Experimental Biology, vol. 307, p. 668-698.; Faria et al., 2011Faria, SC., Augusto, AS. and McNamara, JC., 2011. Intra- and
extracellular osmotic regulation in the hololimnetic Caridea and Anomura: a
phylogenetic perspective on the conquest of fresh water by the decapod
Crustacea. Journal of Comparative Physiology. B, Biochemical, Systemic, and
Environmental Physiology, vol. 181, no. 2, p. 175-186.
http://dx.doi.org/10.1007/s00360-010-0522-6. PMid:20981550
http://dx.doi.org/10.1007/s00360-010-052...
). As more efficient the mechanisms of osmolality
regulation of hemolymph, lower is the necessity of adjustment to the cellular volume
through osmolytes. Both mechanisms involve active transportation through membranes
and, however, adjustment to the respiratory metabolism.
Given that some authors have showed a broad diversity of biological responses among M. amazonicum populations and that this knowledge is still fragmented, the present work has the objective of adding data on the physiology of an estuarine population of M. amazonicum from Amazon, maintained in fish hatcheries for 10 years in the Aquaculture Centre of UNESP, in Jaboticabal, state of São Paulo. We evaluated the effect of M. amazonicum larvae (zoeae I, II, V, IX) and/or post-larvae exposure to different salinities (fresh water, 6, 12 or 18) about the metabolism and/or intracellular isosmotic regulation. The metabolism was assessed by oxygen consumption and intracellular isosmotic regulation through the identification and quantification of body free amino acids.
2 Material and Methods
2.1 Collection, maintenance of the animals in laboratory and larviculture
Macrobrachium amazonicum was obtained from the CAUNESP (UNESP
Aquaculture Centre, Jaboticabal) of the State of São Paulo, Brazil. The
broodstock originated from an estuarine population near Belém in the Amazon
Delta (01°14’30’’S and 48°19’52’’W). Ovigerous females were collected in
reproductive fish hatcheries and kept in a tank of larvae eclosion, in water
with a salinity of 5. After eclosion, larvae were transferred to a polypropylene
tank containing 60 L of fresh water (salinity ≤0.5) or brackish water of 6, 12
or 18 in the density of 100 ind/L. After metamorphosis, post-larvae were
transferred to tanks with capacity of 1000 L to a density of 5 post-larvae/L.
The water temperature in all the experiments was kept at 30°C, photoperiod of
12h:12h of light/dark. Larvae were fed with Artemia
sp nauplii beginning on the second day of cultivation due to
the existence yolk in zoeae I bodies. From the 6th day, humid diet
was added to larvae diet twice a day (Maciel
and Valenti, 2009Maciel, CR. and Valenti, WC., 2009. Biology, fisheries, and
aquaculture of the Amazon River Prawn Macrobrachium amazonicum:
a review. Nauplius, vol. 17, p. 61-79.; Maciel et al.,
2012Maciel, CR., NEW, MB. and Valenti, WC., 2012. The Predation of
artemia nauplii by the larvae of the amazon river prawn, (Heller, 1862), is
affected by prey density, time of day, and ontogenetic development.
Macrobrachium amazonicumJournal of the World Aquaculture
Society, vol. 43, no. 5, p. 659-669.
http://dx.doi.org/10.1111/j.1749-7345.2012.00599.x.
http://dx.doi.org/10.1111/j.1749-7345.20...
). Salinity was daily checked using a refractometer (Atago
S/Mill-E), as well as siphonage of the tanks.
All the ontogenetic stages here studied were selected for the evaluation of the physiological parameters 24h after the change of stage. Thus, all the stages remained at least 24h in experimental salinities (fresh water, 6, 12 or 18). After this period, larvae and post-larvae were withdrawn from the tanks and metabolism and concentration of free amino acids in the body were determined. It was not possible to evaluate the concentration of free amino acids in M. amazonicum zoeae IX because most of the zoeae VIII of our experiments underwent metamorphosis directly to post-larvae; it was possible to obtain enough material for analyses. It was not possible either the determination of oxygen consumption in post-larvae because their elevated oxygen consumption makes the oxygen level within the chambers get to below 70% of saturation, a value that could alter physiological functions and could not be representative.
2.2 Evaluation of the oxygen consumption
Oxygen consumption in zoeae and post-larvae was assessed using a high precision respirometry system for aquatic animals (Strathkelvin Instruments). This system is formed by digital monitor (Mod. 782), respirometric chamber (Mod. MT200), and electrode with micro cathode sensitive to variations in the oxygen dissolved in water, with precision of 0.01 μg/mL (Mod. 1302). The temperature within the respirometric chamber was kept at 30°C with help from a water flow, around the chamber, from thermostatic bath equipment (Tecnal).
The number of larvae transferred to the respirometric chamber in each analysis was defined after a series of preliminary tests using the respirometry system and varied from five (zoeae I) to one (zoeae V, IX) individuals. All the evaluated animals had empty digestive tube. Larvae were acclimated to the respirometric chamber for 30 minutes. After this period, the value of oxygen concentration within the chamber was registered, 60 minutes after a new measure was registered. The difference among values was used in the calculation of individual oxygen consumption. Readings in respirometric chambers with no animals inside were performed, following the same experimental conditions, which were used as control. Variations in oxygen concentration observed in the control chambers were subtracted from the values obtained in respiratory measures of the animals.
After the determination of the oxygen consumption, the animals were killed by freezing, dried at 60°C for 48h (Nova Ética, 400-6ND-200C) and weighted (dry mass). All weight measurement was performed in analytical scale with precision of 1 μg (Mettler). Oxygen consumption is expressed according to the dry mass (μgO2. mgMS–1 h–1).
2.3 Quantification and identification of the free amino acids by hplc
For FAA analyses, the dried tissue samples were homogenized in distilled water,
protein being precipitated with 80% ethanol (v/v). An internal standard of 6.24
nmol α-aminobutyric acid was added and the samples were derivatized with
triethylamine and phenylisothiocyanate, forming FAA/phenylthiocarbamil
derivatives (Bidlingmeyer et al., 1987Bidlingmeyer, BA., Cohen, SA., Tarvin, TL. and Frost, B., 1987. A
new, rapid, high-sensitivity analysis of amino acids in food type samples.
Journal - Association of Official Analytical Chemists, vol. 70, no. 2, p.
241-247. PMid:3571118.).
The individual FAA were identified and quantified by HPLC (Milton Roy) using a
Picotag C18 Column (Waters Corporation) according to Augusto et al. (2007aAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
, bAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007b.
Adaptative shifts in osmoregulatory strategy and the invasion of fresh water by
brachyuran crabs: evidence from . Dilocarcinus pagei
(Trichodactylidae)Journal of Experimental Zoology. Part A, Comparative
Experimental Biology, vol. 307, p. 668-698.).
2.4 Statistical analysis
The effects of exposure to the different salinities on oxygen consumption or FAA concentration were evaluated using 1-way ANOVA followed by the Student-Newman- Keuls multiple means test to locate statistically significant groups. All statistical analyses were performed after determining normality of distribution and equality of variance using Sigma Stat 2.03, employing a minimum significance level of P = 0.05. Data are expressed in the text as mean ± SE.
3 Results
3.1 Survival and duration of larval stages
The exposure of the M. amazonicum larval stages to different salinities revealed differences in what concerns to the survival and duration of the larval development. Whereas in brackish water of 6, 12 and 18 occurred ecdysis in larvae and change the stage until metamorphosis to post-larvae, in fresh water (salinity ≤ 0.5), does no occur ecdysis in the zoea V to zoeae VI. In relation to larval development, we observed an accentuated suppression of zoeae IX stages in all salinities (6, 12 and 18), given that the majority of zoeae VIII underwent metamorphosis directly to post-larvae. The duration of larval development until reaching metamorphosis also varied according to salinity, once on brackish water of 12, larvae suffered metamorphosis in only 17 days, but it took 18 days in brackish water of 6 and 20 days in the salinity of 18.
The accentuated suppression of larval IX stage did not enable the evaluation of the concentration of free amino acids in this M. amazonicum stage due to the reduced quantity of available material. However, it was possible to successfully measure the metabolism of zoeae IX because it is only necessary one individual of this stage in the respirometric chamber to assess the oxygen consumption.
3.2 Metabolism of the selected ontogenetic stages of M. amazonicum
Oxygen consumption of M. amazonicum zoeae I, II, V and IX is shown in Figure 1. In zoeae I, oxygen consumption did not alter due to the exposure salinity. In zoeae II, oxygen consumption increased in the salinity of 18 in comparison to the larvae kept in fresh water. In contrast, in zoeae V, oxygen consumption was higher in zoeae kept in fresh water in comparison to the ones kept in brackish water of 12 or 18. Oxygen consumption was also measured in few larvae that went through the zoea IX stage and we observed that there are no differences in the metabolism of larvae kept in brackish water of 6, 12 or 18. In this stage, there are no data in fresh water because from the zoeae VI stage, all larvae die when kept in this salinity.
Effect of exposure to freshwater or dilute seawater (6, 12 or 18) on oxygen consumption (µg. mg dry weight–1. h–1) in zoea I, II, V and IX of the shrimp Macrobrachium amazonicum. Means in each stage followed by different letters differ statistically. (X ± SEM, N = 4).
3.3 Quantification and identification of free amino acids of the selected ontogenetic stages of M. amazonicum
The total concentration of free amino acids presented variation only in zoeae I and post-larvae (Figure 2). In zoeae I, the total concentration of free amino acids increased from 139.0 ± 15.7 nmol/mg dry mass in fresh water to about 300 nmol/mg dry mass in brackish water of 6, 12 or 18 S. In zoeae II and V, the concentration of free amino acids remained unaltered after the exposure to different salinities and varied from about 320 nmol/mg dry mass in zoeae II in fresh water until about 460 nmol/mg dry mass in zoeae V exposed to brackish water of 18. In post-larvae, the total concentration of free amino acids increased about 40% after exposure to salinity of 18 (553.4 ± 51.6 nmol/mg dry mass) in comparison to those exposed to brackish water of 6 or 12 (respectively, 395.6 ± 9.5 and 405.4 ± 30.7 nmol/mg dry mass).
Effect of exposure to freshwater or dilute seawater (6, 12 or 18) on total free amino acid concentration (nmoles/mg dry weight) in zoea I, II, V and post larvae of the shrimp Macrobrachium amazonicum. Means in each stage followed by different letters differ statistically. (X ± SEM, N = 4).
In all evaluated ontogenetic stages, the most concentrated free amino acids were glutamic acid, glycine, alanine, arginine, and proline together they correspond to about 40% out of the total (Table 1). The exposure of zoeae I to brackish water caused increases in glycine concentration (about 40%), alanine (about 250%), and proline (about 500%) in relation to zoeae kept in fresh water. In zoeae II, the main free amino acids did not have their concentration altered. In zoeae V, proline is the main free amino acid that had its concentration altered, increasing about 200% in larvae exposed to brackish water of 6, 12 and 18 in relation to those kept in fresh water. In post-larvae, glycine concentration reduced about 40% and proline increased about 70% in the individuals exposed to brackish water of 12 in relation to other salinities.
Individual and total free amino acid concentrations (nmoles/mg dry weight) in zoeae I, II, V and post larvae of the shrimp Macrobrachium amazonicum in fresh water or exposed to saline media (6, 12 or 18). Means in each amino acid followed by different letters differ statistically from same stage. (X ± SEM, N = 4).
4 Discussion
The results here presented that come from a population from the estuary of the state
of Pará added information concerning to the existence of distinct physiological
responses among M. amazonicum shrimp populations. The majority of
the populations of this caridean shrimp cannot complete their life cycle in fresh
water, independent on the population coming from the Plata bay or northern region
(Vega Perez, 1984Vega Perez, LA., 1984. Desenvolvimento larval de Macrobrachium
heterochirus (Wiegmann, 1836), Macrobrachium amazonicum (Heller, 1862) e
Macrobrachium brasiliense (Heller,1868) (Crustacea, Decapoda, Palaemonidae), em
laboratório. São Paulo: Universidade de São Paulo, Instituto Oceanográfico. 277
p. Tese de doutorado em Oceanografia.; Zanders et al., 1992;
Augusto et al., 2007aAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
; Charmantier and Anger, 2011Charmantier, G. and Anger, K., 2011. Ontogeny of osmoregulatory
patterns in the South American shrimp . Macrobrachium
amazonicum: loss of hypo-regulation in a land-locked population
indicates phylogenetic separation from estuarine ancestorsJournal of
Experimental Marine Biology and Ecology, vol. 396, no. 2, p. 89-98.
http://dx.doi.org/10.1016/j.jembe.2010.10.013.
http://dx.doi.org/10.1016/j.jembe.2010.1...
). Interestingly,
when maintained in the laboratory, estuarine population here studied survives until
the zoeae V stage in fresh water, whereas in some distinct marine populations, zoeae
die right after eclosion in fresh water (Augusto et
al., 2007aAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
) or do not get to the zoeae III stage if kept in this salinity
(McNamara et al., 1983Mcnamara, JC., Moreira, GS. and Moreira, PS., 1983. The effect of
salinity on respiratory metabolism, survival and moulting in the first zoea of
(Heller) (Crustacea, Palaemonidae). Macrobrachium
amazonicumHydrobiologia, vol. 101, no. 3, p. 239-242.
http://dx.doi.org/10.1007/BF00009880.
http://dx.doi.org/10.1007/BF00009880...
; Gamba, 1984Gamba, AL., 1984. Different egg-associated and larval development
characteristic of and (Arthropoda: Crustacea) in a Venezuelan continental
lagoon. Macrobrachium jelskiiMacrobrachium
amazonicumInternational Journal of Invertebrate Reproduction and
Development. Invertebrate Reproduction & Development, vol. 7, no. 3, p.
135-142. http://dx.doi.org/10.1080/01688170.1984.10510084.
http://dx.doi.org/10.1080/01688170.1984....
). Apparently, from the zoea V
stage, M. amazonicum larvae of the population here studied lose the
capacity to hyperosmoregulate in fresh water, not being able to deal with the water
influx and salts efflux. Charmantier and Anger
(2011)Charmantier, G. and Anger, K., 2011. Ontogeny of osmoregulatory
patterns in the South American shrimp . Macrobrachium
amazonicum: loss of hypo-regulation in a land-locked population
indicates phylogenetic separation from estuarine ancestorsJournal of
Experimental Marine Biology and Ecology, vol. 396, no. 2, p. 89-98.
http://dx.doi.org/10.1016/j.jembe.2010.10.013.
http://dx.doi.org/10.1016/j.jembe.2010.1...
verified that all the post-embryonic stages of the population here
studied are able to hyperosmoregulate until the salinity of 17, although in their
experiments only zoeae I had survived in fresh water. It is possible that different
methodological details used in larvae culture might have caused this imbalance among
the results here presented and the ones from Charmantier and Anger (2011)Charmantier, G. and Anger, K., 2011. Ontogeny of osmoregulatory
patterns in the South American shrimp . Macrobrachium
amazonicum: loss of hypo-regulation in a land-locked population
indicates phylogenetic separation from estuarine ancestorsJournal of
Experimental Marine Biology and Ecology, vol. 396, no. 2, p. 89-98.
http://dx.doi.org/10.1016/j.jembe.2010.10.013.
http://dx.doi.org/10.1016/j.jembe.2010.1...
regarding zoeae survival in fresh
water.
Macrobrachium amazonicum have between 9 and 11 larval stages under
laboratorial conditions in which salinity and temperature are kept, respectively,
around at 10 and 29°C (Guest and Durocher,
1979Guest, WC. and Durocher, PP., 1979. Palaemonid shrimp, .
Macrobrachium amazonicum: effects of salinity and
temperature on survivalProgressive Fish-Culturist, vol. 41, no. 1, p. 14-18.
http://dx.doi.org/10.1577/1548-8659(1979)41[14:PSMA]2.0.CO;2.
http://dx.doi.org/10.1577/1548-8659(1979...
; Vega Perez, 1984Vega Perez, LA., 1984. Desenvolvimento larval de Macrobrachium
heterochirus (Wiegmann, 1836), Macrobrachium amazonicum (Heller, 1862) e
Macrobrachium brasiliense (Heller,1868) (Crustacea, Decapoda, Palaemonidae), em
laboratório. São Paulo: Universidade de São Paulo, Instituto Oceanográfico. 277
p. Tese de doutorado em Oceanografia.; Anger and Hayd, 2009Anger, K. and Hayd, L., 2009. From lecithotrophy to planktotrophy:
ontogeny of larval feeding in the Amazon River prawn Macrobrachium
amazonicum.Aquatic Biology, vol. 7, no. 1-2, p. 19-30.
http://dx.doi.org/10.3354/ab00180.
http://dx.doi.org/10.3354/ab00180...
). We observed in the
present work an abbreviation of the larval stage once almost all zoeae VIII kept on
brackish water of 6, 12 or 18 underwent metamorphosis directly to post-larvae.
Diverse works have shown that caridean shrimps under unfavorable conditions, for
example, during osmotic or nutritional stress, high temperatures, tend to increase
the number of stages or prolonged the duration of larval development (Criales and Anger, 1986Criales, MM. and Anger, K., 1986. Experimental studies on the larval
development of the shrimps and Crangon crangonCrangon
allmanni.Helgolaender Meeresuntersuchungen, vol. 40, no. 3, p.
241-265. http://dx.doi.org/10.1007/BF01983735.
http://dx.doi.org/10.1007/BF01983735...
; Mascetti and Wehrtmann, 1996Mascetti, P. and WEHRTMANN, IS., 1996. Aspects of the reproductive
biology of (Guerin, 1835) (Decapoda, Anomura, Porcellanidae). 3. Effects of
starvation and different types of diet on larval development under laboratory
conditions. Petrolisthes laevigatusArchiv fuer Fischerei und
Meeresforschung, vol. 43, no. 2, p. 159-170.). Low salinity may cause a
reduction of the average rate of feeding or growth efficiency due to metabolic
disadjustments caused by osmotic stress. However, it can be inferred that the
cultivation media in which larvae developed in the present work as salinity,
temperature, and feeding was appropriate to the studied population and it is
reflected in the reduction of the number of larval stages.
Data presented here suggest that possible adjustments in the respiratory metabolism
and in the use of free amino acids in the regulation of larval cellular volume are
dependent on the appearance of structures responsible for osmoregulation and its
functioning. It is known that as more efficient the regulation mechanisms of volume
and composition of extracellular fluid lower the necessity of osmotic effectors in
the regulation of cellular volume. We verified that M. amazonicum
zoeae I did not alter their respiratory metabolism due to exposure to fresh or
brackish water, but reduce the intracellular concentration of free amino acids when
exposed to fresh water, what may suggest the inexistence or an inefficient
performance of the structures responsible for volume regulation and hemolymph
composition. It is even possible that it may be due to a higher permeability of
larvae than other ontogenetic stages to salts and water. The reduction in the
concentration of free amino acids up against the exposure to a diluted media is
related in various crustaceans as Palaemon northropi (Augusto et al., 2009AUGUSTO, A., SILVA, AP., GREENE, LJ., LAURIE, HJ. and MCNAMARA, CM.,
2009. Evolutionary transition to freshwater by ancestral marine palaemonids:
evidence from osmoregulation in a tide pool shrimp. Aquatic Biology, vol. 7, no.
1-2, p. 113-122. http://dx.doi.org/10.3354/ab00183.
http://dx.doi.org/10.3354/ab00183...
) and Litopenaeus
vannamei (Shinji et al., 2012Shinji, J., Okutsu, T., Jayasankar, V., Jasmani, S. and Wilder, MN.,
2012. Metabolism of amino acids during hyposmotic adaptation in the whiteleg
shrimp, Litopenaeus vannamei. Amino Acids, vol. 43, no. 5, p. 1945-1954.
http://dx.doi.org/10.1007/s00726-012-1266-2. PMid:22418866
http://dx.doi.org/10.1007/s00726-012-126...
)
and it is responsible for the maintenance of isosmotic intracellular media with
surrounding extracellular media. This adjustment in the free amino acids
concentration may occur through the reduction of the synthesis rate or increase in
the oxidation of these compounds, increase of amino acid efflux, increase in the
synthesis of proteins or reduction of their catabolism (Mantel and Farmer, 1983Mantel, LH. and Farmer, LL.,1983. Osmotic and ionic regulation. In
MANTEL, LH. (Ed.). Internal anatomy and physiological regulation. New York:
Academic Press. p. 53-161. The Biology of Crustacea, vol. 5.
http://dx.doi.org/10.1016/B978-0-12-106405-1.50013-8.
http://dx.doi.org/10.1016/B978-0-12-1064...
). Hemolymph amino acids increase on
exposure to dilute media and may be stored like hemocyanin, which may increase the
amount of oxygen available to cells (Gilles and
Péqueux, 1981Gilles, R. and Péqueux, A., 1981. Cell volume regulation in
crustaceans: relationship between mechanisms for controlling the osmolality of
extracellular and intracellular fluids. The Journal of Experimental Zoology,
vol. 215, no. 3, p. 351-362.
http://dx.doi.org/10.1002/jez.1402150312.
http://dx.doi.org/10.1002/jez.1402150312...
; Mantel and Farmer,
1983Mantel, LH. and Farmer, LL.,1983. Osmotic and ionic regulation. In
MANTEL, LH. (Ed.). Internal anatomy and physiological regulation. New York:
Academic Press. p. 53-161. The Biology of Crustacea, vol. 5.
http://dx.doi.org/10.1016/B978-0-12-106405-1.50013-8.
http://dx.doi.org/10.1016/B978-0-12-1064...
). Shinji et al. (2012)Shinji, J., Okutsu, T., Jayasankar, V., Jasmani, S. and Wilder, MN.,
2012. Metabolism of amino acids during hyposmotic adaptation in the whiteleg
shrimp, Litopenaeus vannamei. Amino Acids, vol. 43, no. 5, p. 1945-1954.
http://dx.doi.org/10.1007/s00726-012-1266-2. PMid:22418866
http://dx.doi.org/10.1007/s00726-012-126...
and
Shinji and Wilder (2012)Shinji, J. and Wilder, MN., 2012. Dynamics of free amino acids in
the hemolymph of Pacific white leg shrimp . Litopenaeus
vannamei exposed to different types of stressFisheries Science,
vol. 78, no. 6, p. 1187-1194.
http://dx.doi.org/10.1007/s12562-012-0542-0.
http://dx.doi.org/10.1007/s12562-012-054...
verified still
that in adult L.vannamei amino acids were consumed as an energy
source when animals were exposed to low salinity.
In M. amazonicum zoeae II and V, the alterations in the metabolism
during exposure to fresh or brackish water were not followed by alterations in free
amino acids concentration. Thus it is possible that as the structures responsible
for osmoregulation and ionic regulation become effectively functional in larvae, the
role of free amino acids gets diminished and oxygen consumption more elevated,
probably because of the highest energy expenditure with the active transportation of
salts through epithelial membranes. It is possible that gills or other structure
present in initial phases of the development of crustaceans as branchiostegites,
pleurae or dorsal organ are performing osmoregulatory functions, what would reduce
the role of free amino acids in the intracellular volume regulation. The
participation of these structures in the osmotic regulation is cited during the
development of diverse crustaceans as Callianassa jamaicense (Felder et al., 1986Felder, JMD., Felder, L. and Hand, SC., 1986. Ontogeny of
osmoregulation in the estuarine ghost shrimp louisianensis Schmitt (Decapoda,
Thalassinidea). Callianassa jamaicenseJournal of Experimental
Marine Biology and Ecology, vol. 99, no. 2, p. 91-105.
http://dx.doi.org/10.1016/0022-0981(86)90230-3.
http://dx.doi.org/10.1016/0022-0981(86)9...
) and Carcinus
maenas (Cieluch et al.,
2004Cieluch, U., Anger, K., Aujoulat, F., Buchholz, F.,
Charmantier-Daures, M. and Charmantie, G., 2004. Ontogeny of osmoregulatory
structures and functions in the green crab . Carcinus maenas
(Crustacea, Decapoda)The Journal of Experimental Biology, vol. 207, no. Pt 2, p.
325-336. http://dx.doi.org/10.1242/jeb.00759. PMid:14668316
http://dx.doi.org/10.1242/jeb.00759...
).
The osmotic challenges seem to alter throughout the development given that in zoeae
II oxygen consumption is elevated in brackish water of 18, but in zoeae V this
increase happens in fresh water. In addition, from zoeae VI stage, larvae become
incapable of surviving in fresh water and zoeae IX keep unaltered metabolism on
brackish water of 6, 12 or 18. In a M. amazonicum population from
Sertãozinho, Augusto et al. (2007a)Augusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
verified
that only M. amazonicum zoeae I alter their concentration of free
amino acids after exposure to brackish water; the authors attribute it to the
development of structure responsible for the regulation of extracellular fluid in
zoeae II, which would make the osmolality regulation of hemolymph more efficient,
reducing the role of the mechanisms of intracellular isosmotic regulation in this
and in the next phases of development. In M. olfersii diadromous
shrimp, free amino acids concentration did not alter either in zoeae II, although
zoeae I present an increase after exposure to brackish water (Augusto et al., 2007aAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
).
After metamorphosis of M. amazonicum, free amino acids begin to play
an important role as intracellular osmolytes because we verified an increase of up
to 40% in post-larvae of the population here studied exposed to brackish water of
18. These data are similar to the ones presented by the M. olfersii
shrimp post-larvae that present an increase of 28% in the concentration of free
amino acids after the exposure to brackish water (Augusto et al., 2007aAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
). Diadromous shrimp post-larvae of the
Macrobrachium gender constitute the phase of the life cycle
that marks the return to fresh water after larval development in brackish water.
Given that M. amazonicum possess from estuarian to freshwater
populations distant from the sea, the pattern of dependence on brackish water and
migration of the specie also seem to be dependent on their geographical
localization.
The main free amino acids involved in the regulation of cellular volume of the
ontogenetic stages of the M. amazonicum population here studied
were the non essential glutamic acid, glycine, alanine, arginine, and proline. These
amino acids together constitute about 60% of the total of free amino acids,
independent on the ontogenetic stage evaluated. The results obtained here are in
agreement with the literature that shows that the main amino acids which take part
in the osmoregulatory process of crustaceans are the non-essential ones, that means,
those obtained through a biosynthetic via and not trough feeding (McNamara et al., 2004Mcnamara, JC., Rosa, JC., Greene, LJ. and Augusto, A., 2004. Free
amino acid pools as effectors of osmotic adjustment in different tissues of the
freshwater shrimp . Macrobrachium olfersii (Crustacea,
Decapoda) during long-term salinity acclimationMarine and Freshwater Behaviour
and Physiology, vol. 37, no. 3, p. 193-208.
http://dx.doi.org/10.1080/10236240400006208.
http://dx.doi.org/10.1080/10236240400006...
; Augusto et al., 2007aAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The
ontogeny of isosmotic intracellular regulation in the diadromous, freshwater
palaemonid shrimps, and . Macrobrachium amazonicumM.
olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p.
626-634. http://dx.doi.org/10.1651/S-2796.1.
http://dx.doi.org/10.1651/S-2796.1...
, bAugusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007b.
Adaptative shifts in osmoregulatory strategy and the invasion of fresh water by
brachyuran crabs: evidence from . Dilocarcinus pagei
(Trichodactylidae)Journal of Experimental Zoology. Part A, Comparative
Experimental Biology, vol. 307, p. 668-698.; Augusto et al., 2009AUGUSTO, A., SILVA, AP., GREENE, LJ., LAURIE, HJ. and MCNAMARA, CM.,
2009. Evolutionary transition to freshwater by ancestral marine palaemonids:
evidence from osmoregulation in a tide pool shrimp. Aquatic Biology, vol. 7, no.
1-2, p. 113-122. http://dx.doi.org/10.3354/ab00183.
http://dx.doi.org/10.3354/ab00183...
; Faria et al., 2011Faria, SC., Augusto, AS. and McNamara, JC., 2011. Intra- and
extracellular osmotic regulation in the hololimnetic Caridea and Anomura: a
phylogenetic perspective on the conquest of fresh water by the decapod
Crustacea. Journal of Comparative Physiology. B, Biochemical, Systemic, and
Environmental Physiology, vol. 181, no. 2, p. 175-186.
http://dx.doi.org/10.1007/s00360-010-0522-6. PMid:20981550
http://dx.doi.org/10.1007/s00360-010-052...
; Prymaczok et al., 2012Prymaczok, NC., Chaulet, A., Medesani, DA. and Rodríguez, EM., 2012.
Survival, growth, and physiological responses of advanced juvenile freshwater
crayfish (), reared at low temperature and high salinities. Cherax
quadricarinatusAquaculture (Amsterdam, Netherlands), vol. 334-337,
p. 176-181.
http://dx.doi.org/10.1016/j.aquaculture.2011.12.032.
http://dx.doi.org/10.1016/j.aquaculture....
; Shinji et al., 2012Shinji, J., Okutsu, T., Jayasankar, V., Jasmani, S. and Wilder, MN.,
2012. Metabolism of amino acids during hyposmotic adaptation in the whiteleg
shrimp, Litopenaeus vannamei. Amino Acids, vol. 43, no. 5, p. 1945-1954.
http://dx.doi.org/10.1007/s00726-012-1266-2. PMid:22418866
http://dx.doi.org/10.1007/s00726-012-126...
).
The results presented here broaden the range of physiological responses described for
M. amazonicum populations, because larvae can survive until
zoeae V stage in fresh water and only some stages possess dependence on free amino
acids in the cellular volume regulation and adjust their metabolism according to
exposure to brackish water. This pattern is different from the one found by some
M. amazonicum populations where larvae die in fresh water right
after eclosion or in other initial stages of the life cycle. Additionally, data add
useful information to aquaculture of M. amazonicum, considered
promising specie for commercial cultivation (New et
al., 2010New, MB., Valenti, WC., Tidwell, JH., Abramo, LRD. and Kutty, MN.,
2010. Freshwater Prawns Biology and Farming. Oxford: Wiley-Blackwell. 544
p.; Bentes et al., 2011BENTES, BS., MARTINELLI, JM., SOUZA, LS., CAVALCANTE, DV., ALMEIDA,
MC. and ISAAC, VJ., 2011. Spatial distribution of the amazon river prawn
Macrobrachium amazonicum (Heller, 1862) (Decapoda, Caridea, Palaemonidae) in two
perennial creeks of an estuary on the northern coast of Brazil (Guajara Bay,
Belem, Para). Brazilian Journal of Biology, vol. 71, no. 4, p. 925-935.
http://dx.doi.org/10.1590/S1519-69842011000500013.
http://dx.doi.org/10.1590/S1519-69842011...
).
The fact that zoeae II canalize greater quantity of energy for osmoregulatory
processes when kept in brackish water of 18 may negatively influence their growth
and development. Taking into account only metabolic parameters, it would be
interesting that the larvae cultivated for commercial purposes were kept in fresh
water or brackish water of up to 12 until zoeae II stage.
Acknowledgements
We thank the Carciniculture Laboratory of the Aquaculture Center of UNESP (CAUNESP) in the person of its coordinator Dr Wagner Cotroni Valenti for all the support offered during the time experiments were conducted. The authors are grateful to the Protein Chemistry Center of the Hemocentro/USP/RP by the collaboration in the free amino acids analysis by HPLC. Alessandra Augusto received grants from FAPESP (process 07/56553-7) and CNPQ (process 479428/2009-3).
References
- Anger, K. and Hayd, L., 2009. From lecithotrophy to planktotrophy: ontogeny of larval feeding in the Amazon River prawn Macrobrachium amazonicum.Aquatic Biology, vol. 7, no. 1-2, p. 19-30. http://dx.doi.org/10.3354/ab00180.
» http://dx.doi.org/10.3354/ab00180 - Augusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007a. The ontogeny of isosmotic intracellular regulation in the diadromous, freshwater palaemonid shrimps, and . Macrobrachium amazonicumM. olfersi (Decapoda)Journal of Crustacean Biology, vol. 27, no. 4, p. 626-634. http://dx.doi.org/10.1651/S-2796.1.
» http://dx.doi.org/10.1651/S-2796.1 - Augusto, A., Greene, LJ., Laure, HJ. and McNamara, JC., 2007b. Adaptative shifts in osmoregulatory strategy and the invasion of fresh water by brachyuran crabs: evidence from . Dilocarcinus pagei (Trichodactylidae)Journal of Experimental Zoology. Part A, Comparative Experimental Biology, vol. 307, p. 668-698.
- AUGUSTO, A., SILVA, AP., GREENE, LJ., LAURIE, HJ. and MCNAMARA, CM., 2009. Evolutionary transition to freshwater by ancestral marine palaemonids: evidence from osmoregulation in a tide pool shrimp. Aquatic Biology, vol. 7, no. 1-2, p. 113-122. http://dx.doi.org/10.3354/ab00183.
» http://dx.doi.org/10.3354/ab00183 - BENTES, BS., MARTINELLI, JM., SOUZA, LS., CAVALCANTE, DV., ALMEIDA, MC. and ISAAC, VJ., 2011. Spatial distribution of the amazon river prawn Macrobrachium amazonicum (Heller, 1862) (Decapoda, Caridea, Palaemonidae) in two perennial creeks of an estuary on the northern coast of Brazil (Guajara Bay, Belem, Para). Brazilian Journal of Biology, vol. 71, no. 4, p. 925-935. http://dx.doi.org/10.1590/S1519-69842011000500013.
» http://dx.doi.org/10.1590/S1519-69842011000500013 - Bidlingmeyer, BA., Cohen, SA., Tarvin, TL. and Frost, B., 1987. A new, rapid, high-sensitivity analysis of amino acids in food type samples. Journal - Association of Official Analytical Chemists, vol. 70, no. 2, p. 241-247. PMid:3571118.
- Charmantier, G. and Anger, K., 2011. Ontogeny of osmoregulatory patterns in the South American shrimp . Macrobrachium amazonicum: loss of hypo-regulation in a land-locked population indicates phylogenetic separation from estuarine ancestorsJournal of Experimental Marine Biology and Ecology, vol. 396, no. 2, p. 89-98. http://dx.doi.org/10.1016/j.jembe.2010.10.013.
» http://dx.doi.org/10.1016/j.jembe.2010.10.013 - Cieluch, U., Anger, K., Aujoulat, F., Buchholz, F., Charmantier-Daures, M. and Charmantie, G., 2004. Ontogeny of osmoregulatory structures and functions in the green crab . Carcinus maenas (Crustacea, Decapoda)The Journal of Experimental Biology, vol. 207, no. Pt 2, p. 325-336. http://dx.doi.org/10.1242/jeb.00759. PMid:14668316
» http://dx.doi.org/10.1242/jeb.00759 - Criales, MM. and Anger, K., 1986. Experimental studies on the larval development of the shrimps and Crangon crangonCrangon allmanni.Helgolaender Meeresuntersuchungen, vol. 40, no. 3, p. 241-265. http://dx.doi.org/10.1007/BF01983735.
» http://dx.doi.org/10.1007/BF01983735 - Faria, SC., Augusto, AS. and McNamara, JC., 2011. Intra- and extracellular osmotic regulation in the hololimnetic Caridea and Anomura: a phylogenetic perspective on the conquest of fresh water by the decapod Crustacea. Journal of Comparative Physiology. B, Biochemical, Systemic, and Environmental Physiology, vol. 181, no. 2, p. 175-186. http://dx.doi.org/10.1007/s00360-010-0522-6. PMid:20981550
» http://dx.doi.org/10.1007/s00360-010-0522-6 - Felder, JMD., Felder, L. and Hand, SC., 1986. Ontogeny of osmoregulation in the estuarine ghost shrimp louisianensis Schmitt (Decapoda, Thalassinidea). Callianassa jamaicenseJournal of Experimental Marine Biology and Ecology, vol. 99, no. 2, p. 91-105. http://dx.doi.org/10.1016/0022-0981(86)90230-3.
» http://dx.doi.org/10.1016/0022-0981(86)90230-3 - Freire, CA., Onken, H. and McNamara, JC., 2008. A structure-function analysis of ion transport in crustacean gills and excretory organs. Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology, vol. 151, no. 3, p. 272-304. http://dx.doi.org/10.1016/j.cbpa.2007.05.008. PMid:17604200
» http://dx.doi.org/10.1016/j.cbpa.2007.05.008 - Gamba, AL., 1984. Different egg-associated and larval development characteristic of and (Arthropoda: Crustacea) in a Venezuelan continental lagoon. Macrobrachium jelskiiMacrobrachium amazonicumInternational Journal of Invertebrate Reproduction and Development. Invertebrate Reproduction & Development, vol. 7, no. 3, p. 135-142. http://dx.doi.org/10.1080/01688170.1984.10510084.
» http://dx.doi.org/10.1080/01688170.1984.10510084 - Garçon, DP., Lucena, MN., Pinto, MR., Fontes, CFL., McNamara, JC. and Leone, FA., 2013. Synergistic stimulation by potassium and ammonium of K(+)-phosphatase activity in gill microsomes from the crab Callinectes ornatus acclimated to low salinity: novel property of a primordial pump. Archives of Biochemistry and Biophysics, vol. 530, no. 2, p. 55-63. http://dx.doi.org/10.1016/j.abb.2012.12.006. PMid:23262318
» http://dx.doi.org/10.1016/j.abb.2012.12.006 - Gilles, R. and Péqueux, A., 1981. Cell volume regulation in crustaceans: relationship between mechanisms for controlling the osmolality of extracellular and intracellular fluids. The Journal of Experimental Zoology, vol. 215, no. 3, p. 351-362. http://dx.doi.org/10.1002/jez.1402150312.
» http://dx.doi.org/10.1002/jez.1402150312 - Guest, WC. and Durocher, PP., 1979. Palaemonid shrimp, . Macrobrachium amazonicum: effects of salinity and temperature on survivalProgressive Fish-Culturist, vol. 41, no. 1, p. 14-18. http://dx.doi.org/10.1577/1548-8659(1979)41[14:PSMA]2.0.CO;2.
» http://dx.doi.org/10.1577/1548-8659(1979)41[14:PSMA]2.0.CO;2 - Maciel, CR. and Valenti, WC., 2009. Biology, fisheries, and aquaculture of the Amazon River Prawn Macrobrachium amazonicum: a review. Nauplius, vol. 17, p. 61-79.
- Maciel, CR., NEW, MB. and Valenti, WC., 2012. The Predation of artemia nauplii by the larvae of the amazon river prawn, (Heller, 1862), is affected by prey density, time of day, and ontogenetic development. Macrobrachium amazonicumJournal of the World Aquaculture Society, vol. 43, no. 5, p. 659-669. http://dx.doi.org/10.1111/j.1749-7345.2012.00599.x.
» http://dx.doi.org/10.1111/j.1749-7345.2012.00599.x - Mantel, LH. and Farmer, LL.,1983. Osmotic and ionic regulation. In MANTEL, LH. (Ed.). Internal anatomy and physiological regulation. New York: Academic Press. p. 53-161. The Biology of Crustacea, vol. 5. http://dx.doi.org/10.1016/B978-0-12-106405-1.50013-8.
» http://dx.doi.org/10.1016/B978-0-12-106405-1.50013-8 - Mascetti, P. and WEHRTMANN, IS., 1996. Aspects of the reproductive biology of (Guerin, 1835) (Decapoda, Anomura, Porcellanidae). 3. Effects of starvation and different types of diet on larval development under laboratory conditions. Petrolisthes laevigatusArchiv fuer Fischerei und Meeresforschung, vol. 43, no. 2, p. 159-170.
- Mcnamara, JC., Moreira, GS. and Moreira, PS., 1983. The effect of salinity on respiratory metabolism, survival and moulting in the first zoea of (Heller) (Crustacea, Palaemonidae). Macrobrachium amazonicumHydrobiologia, vol. 101, no. 3, p. 239-242. http://dx.doi.org/10.1007/BF00009880.
» http://dx.doi.org/10.1007/BF00009880 - Mcnamara, JC., Rosa, JC., Greene, LJ. and Augusto, A., 2004. Free amino acid pools as effectors of osmotic adjustment in different tissues of the freshwater shrimp . Macrobrachium olfersii (Crustacea, Decapoda) during long-term salinity acclimationMarine and Freshwater Behaviour and Physiology, vol. 37, no. 3, p. 193-208. http://dx.doi.org/10.1080/10236240400006208.
» http://dx.doi.org/10.1080/10236240400006208 - New, MB., Valenti, WC., Tidwell, JH., Abramo, LRD. and Kutty, MN., 2010. Freshwater Prawns Biology and Farming. Oxford: Wiley-Blackwell. 544 p.
- Péqueux, A., 1995. Osmotic regulation in crustaceans. Journal of Crustacean Biology, vol. 15, no. 1, p. 1-60. http://dx.doi.org/10.2307/1549010.
» http://dx.doi.org/10.2307/1549010 - Prymaczok, NC., Chaulet, A., Medesani, DA. and Rodríguez, EM., 2012. Survival, growth, and physiological responses of advanced juvenile freshwater crayfish (), reared at low temperature and high salinities. Cherax quadricarinatusAquaculture (Amsterdam, Netherlands), vol. 334-337, p. 176-181. http://dx.doi.org/10.1016/j.aquaculture.2011.12.032.
» http://dx.doi.org/10.1016/j.aquaculture.2011.12.032 - Shinji, J. and Wilder, MN., 2012. Dynamics of free amino acids in the hemolymph of Pacific white leg shrimp . Litopenaeus vannamei exposed to different types of stressFisheries Science, vol. 78, no. 6, p. 1187-1194. http://dx.doi.org/10.1007/s12562-012-0542-0.
» http://dx.doi.org/10.1007/s12562-012-0542-0 - Shinji, J., Okutsu, T., Jayasankar, V., Jasmani, S. and Wilder, MN., 2012. Metabolism of amino acids during hyposmotic adaptation in the whiteleg shrimp, Litopenaeus vannamei. Amino Acids, vol. 43, no. 5, p. 1945-1954. http://dx.doi.org/10.1007/s00726-012-1266-2. PMid:22418866
» http://dx.doi.org/10.1007/s00726-012-1266-2 - Urzúa, A. and Anger, K., 2011. Larval biomass and chemical composition at hatching in two geographically isolated clades of the shrimp Macrobrachium amazonicum: intra- or interspecific variation?Invertebrate Reproduction & Development, vol. 55, no. 4, p. 236-246. http://dx.doi.org/10.1080/07924259.2011.576155.
» http://dx.doi.org/10.1080/07924259.2011.576155 - Vega Perez, LA., 1984. Desenvolvimento larval de Macrobrachium heterochirus (Wiegmann, 1836), Macrobrachium amazonicum (Heller, 1862) e Macrobrachium brasiliense (Heller,1868) (Crustacea, Decapoda, Palaemonidae), em laboratório. São Paulo: Universidade de São Paulo, Instituto Oceanográfico. 277 p. Tese de doutorado em Oceanografia.
- Vergamini, FG., Pileggi, LG. and Mantelatto, FL., 2011. Genetic variability of the Amazon River prawn (Decapoda, Caridea, Palaemonidae). Macrobrachium amazonicumContributions to Zoology (Amsterdam, Netherlands), vol. 80, no. 1, p. 67-83.
- Zanders, IP. and Rodriguez, JM., 1992. Effect of temperature and salinity stress on osmoionic regulation in adult and on oxygen consumption in larvae and adult of . Macrobrachium amazonicum (Decapoda, Palaemonidae)Comparative Biochemistry and Physiology. A. Comparative Physiology, vol. 101, no. 3, p. 505-509. http://dx.doi.org/10.1016/0300-9629(92)90502-H.
» http://dx.doi.org/10.1016/0300-9629(92)90502-H
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(With 2 figures)
Publication Dates
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Publication in this collection
May 2015
History
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Received
06 Aug 2013 -
Accepted
22 Oct 2013