Phylogeography of the widespread Australian freshwater prawn, Macrobrachium australiense (Decapoda, Palaemonidae)

Aim To investigate the phylogeographic structure of the widespread freshwater prawn, Macrobrachium australiense, within and between major Australian drainage basins using mitochondrial sequence data. This will enable the investigation of historical connections between major drainages and examination of hypotheses of biogeographic associations among Australian freshwater basins. Location Inland, eastern and northern Australia. Methods Sequencing 16S rRNA and ATPase 6 protein coding mitochondrial DNA genes from M. australiense from 19 locations from inland, eastern and northern Australia. Results Within drainage basins, haplotype trees are monophyletic, with the exception of the Finke River from the Lake Eyre Basin. Macrobrachium australiense from the two main inland drainages, the Murray–Darling and Lake Eyre Basin are divergent from each other and do not form a monophyletic group, instead the Murray–Darling Basin haplotypes clade with eastern coastal haplotypes. Haplotypes from neighbouring eastern coastal drainages were found to be quite divergent from each other. Main conclusions The phylogeographic relationships among M. australiense suggest that the two major inland drainages, the Murray–Darling Basin and the Lake Eyre Basin, are not biogeographically closely associated to each other. Instead the Murray–Darling Basin is more closely allied with the eastern coastal drainages across the Great Dividing Range. Despite their proximity the neighbouring southeast Queensland coastal Mary and Brisbane Rivers are also biogeographically divergent from each other. The results also indicate that the Finke River appears to have been isolated from the remainder of the Lake Eyre Basin catchment for a significant period of time.     

Embryonic development of the Caridean prawn Macrobrachium mammillodactylus (Crustacea: Decapoda: Palaemonidae)

The freshwater knob-tooth prawn Macrobrachium mammillodactylus is a commercially exploited species in the Philippines. To study the biology of this species, broodstock from the wild was collected, transported to the laboratory and kept in pairs in indoor polyethylene tanks for breeding. Eggs from berried females were sampled to follow the stages of embryonic development until hatching to zoea larva. It took 18.0?±?2.1?days for the eggs to hatch at ambient water temperature between 25 and 28?°C. The morphological landmarks of development at the different stages (pre-cleavage, cleavage, blastula, pre-nauplius, post-nauplius and pre-hatching) of the live embryos are described. Incremental percentage staging was adopted from 0% at fertilization to 100% at hatching and were matched with corresponding morphological development. Egg volume increased significantly toward the mid-to-later stages of development. The eye index also showed a significant increase as the egg developed. The colour of the egg mass changed from light olive green to grey as the eggs progressed in development. The general pattern of development was comparable to other members of the genus Macrobrachium.     

Phylogenetic relationships of the globally distributed freshwater prawn genus Macrobrachium (Crustacea: Decapoda: Palaemonidae): biogeography, taxonomy and the convergent evolution of abbreviated larval development

There has hitherto been little research into evolutionary and taxonomic relationships amongst species of the freshwater prawn genus Macrobrachium Bate across its global distribution. Previous work by the authors demonstrated that the endemic Australian species did not evolve from a single ancestral lineage. To examine whether other regional Macrobrachium faunas also reflect this pattern of multiple origins, the phylogeny of 30 Macrobrachium species from Asia, Central/South America and Australia was inferred from mitochondrial 16S rRNA sequences. Phylogenetic relationships demonstrate that, despite some evidence for regional diversification, Australia, Asia and South America clearly contain Macrobrachium species that do not share a common ancestry, suggesting that large-scale dispersal has been a major feature of the evolutionary history of the genus. The evolution of abbreviated larval development (ALD), associated with the transition from an estuarine into a purely freshwater lifecycle, was also mapped onto the phylogeny and was shown to be a relatively homoplasious trait and not taxonomically informative. Other taxonomic issues, as well as the evolutionary origins of Macrobrachium , are also discussed.     

A Revision of Australian River Prawns, Macrobrachium (Crustacea: Decapoda: Palaemonidae)

A taxonomic revision of Australian Macrobrachium identified three species new to the Australian fauna – two undescribed species and one new record, viz. M. auratumsp. nov., M. koombooloombasp. nov., and M. mammillodactylus(Thallwitz, 1892). Eight taxa previously described by Riek (1951) are recognised as new junior subjective synonyms, viz. M. adscitum adscitum, M. atactum atactum, M. atactum ischnomorphum, M. atactum sobrinum, M. australiense crassum, M. australiense cristatum, M. australiense eupharum of M. australienseHolthuis, 1950, and M. glypticumof M. handschiniRoux, 1933. Apart from an erroneous type locality for a junior subjective synonym, there were no records to confirm the presence of M. australe(Guérin-Méneville, 1838) on the Australian continent. In total, 13 species of Macrobrachiumare recorded from the Australian continent. Keys to male developmental stages and Australian species are provided. A revised diagnosis is given for the genus. A list of 31 atypical species which do not appear to be based on fully developed males or which require re-evaluation of their generic status is provided. Terminology applied to spines and setae is revised.     

A re-description of Macrobrachium handschini (Roux, 1933) (Crustacea: Decapoda: Palaemonidae)

Diversity, distribution, seasonal changes in density and relative abundance of Ephemeroptera nymphs were studied in five stations on three streams in and around Shillong, Meghalaya state, North-eastern India. Seventeen species belonging to eight genera and five families were recorded. Diversity (Shannon's index) was governed by the heterogeneity of the substratum which in turn was influenced by catchment disturbances (quarrying, logging, and entry of sewage). Seasonal changes in density were governed by the spate-frequency regime of the streams. Relative abundance showed dominance of one or two species at impacted stations, while in less disturbed ones, a more equitable distribution of species was observed.     

Geometric morphometric analysis of the freshwater prawn Macrobrachium borellii (Decapoda: Palaemonidae) at a microgeographical scale in a floodplain system

Morphometric studies are useful for delineating the shapes of various populations and species over geographical ranges and as evidence of regional differences in crustaceans. Hydrological fluctuations in a floodplain system modulate the dispersal and presence of decapods among habitats and constitute an important macrofactor that regulate other environmental variables and which could explain the richness, distribution and abundance of organisms that live in these systems. Morphological variations among populations of the freshwater prawn Macrobrachium borellii in a floodplain system at a microgeographical scale were studied using geometric morphometrics. Carapace structure was represented using 16 digitised landmarks. Allometry and sexual dimorphism was tested. Variation in shape was explored via Principal Component Analysis. Canonical Variate Analyses was applied to compare the differences in shape between species’ populations. The correlations and covariations among shapes and hydrometric level, current velocity, geographical location and hydrologic distances were analysed. The average carapace shape was different between sexes in all sites. Populations that were near each other in terms of hydrological distance had similar shapes, but all of the populations differed in shape from the farthest population. The environmental variables were not good predictors of the carapace shape. Instead, the shape was strongly related to the hydrologic distance and geographical location. The swimming characteristics of these prawns and their passive movements, together with the dynamics of a floodplain system, explain the low morphological variation between populations in this study. The dynamic characteristics of the system influence the dispersal of the prawns and allow populations to remain connected.     

Integrated morphometric and molecular analyses indicate three male morphotypes in the freshwater prawn Macrobrachium olfersii (Decapoda,Palaemonidae) along the Brazilian neotropical region

Males of Macrobrachium olfersii have a large cheliped (second pereiopod) with individual variations in shape and ornamentation, and they vary in size within and between populations. Some misidentification or doubts about taxonomic validity occur due to this morphological variability throughout their geographic distribution. We carried out a morphometric analysis to detect specific patterns and the potential occurrence of morphotypes in M. olfersii from different populations. Our data set included 52 females and 109 males collected in the Neotropical region, between the north and south limits of Brazilian territory, in coastal rivers from the states of Rio Grande do Norte, Bahia, Espírito Santo, Rio de Janeiro, São Paulo, Paraná and Santa Catarina. Body measurements were size standardized by allometric methods and explored by the principal component analysis (PCA) and non-hierarchical analysis of K-means clustering and canonical variate analysis (CVA). Our results suggested three distinct morphological groups (morphotypes) only for males in M. olfersii. Furthermore, the confirmation of monophyly of the different populations of M. olfersii was accomplished by sequences of 16S rDNA. Additional studies about relative growth in other populations and the analysis of the entire ontogenetic development of males in the laboratory will be important to complement the knowledge about morphotypes in this species.     

Trophic ecology of the freshwater prawn, Pseudopalaemon bouvieri (Decapoda: Palaemonidae) in Northeastern Argentina, with remarks on population structure

Freshwater decapod crustaceans are important components of food webs in these environments, but little is known about the diet of species that live in tropical waters. We studied the feeding ecology of the prawn Pseudopalaemon bouvieri and its population structure in two different areas with six lagoons, with a different composition and abundance of aquatic macrophytes. At each site of macrophytes banks, 18 prawns sample was collected with a hand net (1mm mesh size) from 1m2. In the laboratory, prawns cephalothorax length was measured, sex determined, and a total of 208 stomachs were examined for food items. Our results showed that the population abundance varied between 10 ind/m2 and 1 411 ind/m2. The cephalothorax length ranged between 6mm and 21mm, and the male:female ratio varied between 0.3 and 1.0, with a higher proportion of ovigerous females (21%) in area one than area two. P. bouvieri is omnivorous, and its diet was principally based on algae, plant remains, Protozoa, Rotifera, Oligochaeta, Crustacea, Insecta, detritus and other items. The analysis of the stomach content did not reveal any significant difference in the diet between juveniles and adults, and males and females of both areas consumed a similar diet (Kruskal-Wallis test p=0.8273). We concluded that the dietary items consumed by prawns and the niche breadth were similar between the two areas, although the proportion of items consumed varied between lagoons of both areas. The density of P. bouvieri was different between areas, but the size of cephalothorax (CL) was similar.     

Polymorphic microsatellite markers of freshwater prawn Palaemon paucidens (De Haan, 1844) (Crustacea: Palaemonidae)

We report 11 polymorphic microsatellite loci developed from the freshwater prawn Palaemon paucidens. Their genetic characteristics were assessed in 48 individuals selected from six Korean populations. The number of alleles ranged from two to 21, and the observed and expected heterozygosities were between 0.13 and 0.83, and 0.46 and 0.95, respectively. We examined the cross-specific amplification of each locus in three species of palaemonid prawn and one species of atyid prawn.     

Genetic structure and dispersal of Macrobrachium australiense (Decapoda: Palaemonidae) in western Queensland, Australia

SUMMARY 1. The freshwater prawn, Macrobrachium australiense (Decapoda: Palaemonidae), is an abundant species throughout the rivers of western Queensland, and it is thought to have effective dispersal capabilities. Given the very low topographical relief of the western Queensland landscape and the vast nature of episodic flooding, it was predicted that widespread dispersal in this species would occur within, and possibly between the four major catchments of the region: the Darling, Bulloo, Cooper and Diamantina. We analysed eight polymorphic allozyme systems and a fragment of the cytochrome c oxidase subunit 1 (COI) mitochondrial DNA (mtDNA) gene to determine the extent of recent and historical patterns of dispersal at nested spatial scales, within and between catchments.
2. Large and significant levels of allozyme and mtDNA differentiation were revealed among all catchments, indicating that dispersal of M. australiense does not occur across catchment boundaries, although this species is reportedly capable of overland movement. In contrast, no significant patterns of genetic differentiation were resolved between major subcatchments of the Darling and Cooper, or between sites within these subcatchments, indicating that populations of M. australiense are panmictic within catchments.
3. The MtDNA data resolved two divergent and reciprocally monophyletic clades, with the first representing the Darling catchment, and the second corresponding to the Bulloo, Cooper and Diamantina catchments. We postulate that extreme variation in historical climatic patterns and palaeohydrologic conditions played an important role in shaping the population structure of M. australiense throughout western Queensland during the Quaternary.     

Continuous light increases N-acetyltransferase activity in the optic lobe of the giant freshwater prawn Macrobrachium rosenbergii de Man (Crustacea: Decapoda).

Giant freshwater prawns, Macrobrachium rosenbergii de Man, were reared under three different lighting conditions: continuous darkness (DD), 12 hr of light and 12 hr of darkness (LD 12:12) and continuous light (LL). After one month, the prawns were sacrificed and optic lobes isolated from the eyestalks were determined for N-acetyltransferase (NAT) activities and melatonin concentrations. Gonads were weighed and examined under light microscopy. The optic lobes from LL prawns contained significantly higher activities of NAT than those from LD 12:12 prawns. The melatonin concentrations and size and histological features of the gonads from the three groups of prawns did not differ. The results indicate that continuous light increases NAT activities in the optic lobe of M. rosenbergii but has no drastic effect on gonadal growth.     

Na,K-ATPase activity and epithelial interfaces in gills of the freshwater shrimp Macrobrachium amazonicum (Decapoda,Palaemonidae)

Diadromous freshwater shrimps are exposed to brackish water both as an obligatory part of their larval life cycle and during adult reproductive migration; their well-developed osmoregulatory ability is crucial to survival in such habitats. This study examines gill microsomal Na,K-ATPase (K-phosphatase activity) kinetics and protein profiles in the freshwater shrimp Macrobrachium amazonicum when in fresh water and after 10-days of acclimation to brackish water (21‰ salinity), as well as potential routes of Na⁺ uptake across the gill epithelium in fresh water. On acclimation, K-phosphatase activity decreases 2.5-fold, Na,K-ATPase α-subunit expression declines, total protein expression pattern is markedly altered, and enzyme activity becomes redistributed into different density membrane fractions, possibly reflecting altered vesicle trafficking between the plasma membrane and intracellular compartments. Ultrastructural analysis reveals an intimately coupled pillar cell-septal cell architecture and shows that the cell membrane interfaces between the external medium and the hemolymph are greatly augmented by apical pillar cell evaginations and septal cell invaginations, respectively. These findings are discussed regarding the putative movement of Na⁺ across the pillar cell interfaces and into the hemolymph via the septal cells, powered by the Na,K-ATPase located in their invaginations.     

Triacylglycerol catabolism in the prawn Macrobrachium borellii (Crustacea: Palaemoniade)

While invertebrates store neutral lipids as their major energy source, little is known about triacylglycerol (TAG) class composition and their differential catabolism in aquatic arthropods. This study focuses on the composition of the main energy source and its catabolism by lipase from the midgut gland (hepatopancreas) of the crustacean Macrobrachium borellii. Silver-ion thin-layer chromatography of prawn large TAG deposit (80% of total lipids) and its subsequent fatty acid analysis by gas chromatography allowed the identification of 4 major fractions. These are composed of fatty acids of decreasing unsaturation and carbon chain length, the predominant being 18:1n-9. Fraction I, the most unsaturated one, contained mainly 20:5n-3; fraction II 18:2n-6; fraction III 18:1n-9 while the most saturated fraction contained mostly 16:0. Hepatopancreas main lipase (Mr 72 kDa) cross-reacted with polyclonal antibodies against insect lipase, was not dependent on the presence of Ca²⁺ and had an optimum activity at 40 °C and pH 8.0. Kinetic analysis showed a Michaelis–Menten behavior. A substrate competition assay evidenced lipase specificity following the order: 18:1n-9-TAG > PUFA-enriched-TAG > 16:0-TAG different from that in vertebrates. These data indicate there is a reasonable correspondence between the fatty acid composition of TAG and the substrate specificity of lipase, which may be an important factor in determining which fatty acids are mobilized during lipolysis for oxidation in crustaceans.     

Setal morphology of the grooming appendages of Macrobrachium rosenbergii (Crustacea: Decapoda: Caridea: Palaemonidae) and review of decapod setal classification

Setae are vital in grooming activities and aiding in the removal of epibionts and sedimentary fouling from the body surfaces of decapod crustaceans. Thus, the setal structures and their arrangement on the grooming appendages and sensory structures of the commercially important shrimp, Macrobrachium rosenbergii, were examined using scanning electron microscopy. Macrobrachium rosenbergii is extensively grown in aquaculture and exhibits unique male morphological forms, termed morphotypes. The three male morphotypes are termed blue‐clawed males, orange‐clawed males, and small‐clawed or undifferentiated males and all three differ in their dominance, behavior, body morphology, and reproductive success. Seven setal types, two of which have never been described in the literature, are identified on the grooming appendages (third maxillipeds, first, second, and fifth pereopods) and antennae: simple, serrate, serrulate, spiniform, pappose, crinoid, and spinulate. The latter two setae are newly identified. Certain setal types, such as serrate and serrulate setae were located and associated with specific grooming appendages such as the first pereopods. The types of setae on the grooming appendages varied among females and male morphotypes and the novel setal types (crinoid and spinulate) were found only on two of the male morphotypes. A literature review of terminology related to the structure of setae and setal types in decapod crustaceans is offered as the usage of various terms is ambiguous and conflicting in the literature. The intention of this review is to provide future authors with a comprehensive collection of terms and images that can be used to describe various aspects of setal morphology in decapods. J. Morphol. 275:634–649, 2014. © 2014 Wiley Periodicals, Inc.     

Effect of salinity on the embryonic development of Macrobrachium acanthurus (Decapoda: Palaemonidae)

The effect of salinity level on the embryonic development of Macrobrachium acanthurus was analyzed under laboratory conditions, considering characteristics of the egg (size, volume, and water content) and of the embryo (eye index). The experimental design was completely randomized, with five repetitions (ovigerous females) per treatment (0, 10, 17 and 20 ppt). During embryonic development, two eggs per female were taken daily for analyses of size, volume, water content, and eye index. Our results showed that salinity of 20 ppt leads to death and/or abortion of the embryo in all females. The size, volume and water content of eggs increased according to embryonic development, providing space in the egg for formation and organization of embryo. Salinity affected these egg characteristics, causing water loss to the hypertonic medium. Neither the duration of embryonic development nor embryo formation were affected by saltwater content. The results of the present study indicate that ovigerous females of M. acanthurus can survive in freshwater rivers as well as in low-salinity environments during incubation period and the successful larval development is not likely to rely on female migrating to estuaries. Larvae can easily be incubated in freshwater and complete development at higher salinities after hatch.     

Reproductive pattern of the freshwater prawn Pseudopalaemon bouvieri (Crustacea,Palaemonidae) from hypo-osmotic shallow lakes of Corrientes (Argentina)

Pseudopalaemon bouvieri undergoes complete abbreviated development. The reproductive cycle (proportion and size of the sexually mature population and juvenile recruitment), fecundity and egg size of this prawn were examined in three subtropical shallow lakes. The reproductive cycle was seasonal; gonadal maturation occurred during the winter, ovigerous females were abundant in the spring and breeding occurred at the end of spring. Females produced small numbers of eggs (9–55) of relatively large sizes (1.0–2.1 mm). The proportion of ovigerous females with respect to the total number of females, the fecundity and egg size differed among the lakes according to the trophic state of the environments. Pseudopalaemon bouvieri has a reproductive strategy similar to other Palaemonidae species that inhabit nutrient-poor inland waters.     

Morphology of the male reproductive system of freshwater prawn Macrobrachium carcinus (Decapoda,Caridea): Functional and comparative aspects

Testes and vasa deferentia are parts of the male reproductive system of decapod crustaceans. Both organs show morphological differences among decapod species in terms of anatomical and histological patterns reflecting the diversity of this group. Describing these features may assist in systematics, phylogenetics, and studies of reproductive behavior, especially for species of commercial interest, such as Macrobrachium carcinus, a native American species that, unusually for this genus, has no precopulation courting behavior. This study aims to describe the reproductive morphology and spermatogenesis of the male freshwater prawn M. carcinus. The male reproductive system of this species consisted of lobed testes connected to the vasa deferentia. The testis of M. carcinus was divided into several lobules. Each lobule was formed by a cluster of germ cells surrounded by connective tissue and nurse cells. This microscopic anatomy and histology of the testicular histoarchitecture has been described for many species of Decapoda and may represent a derived design of the testes. Unlike that in other decapod species, spermatogenesis proceeds in short transitory phases that produce spermatozoa at high concentrations and frequencies, corroborating the uncommon male reproductive behavior of this species. In the spermatic pathway, the lobules develop and fuse before releasing spermatozoa from the testes; however, this process has not been observed in decapods, yet. The neutral compounds secreted by the vas deferens are important for sperm nutrition as females secrete a substance for spermatophore adhesion during reproduction. This study presents different features and dynamics of the spermatogenic process in the male reproductive system of M. carcinus that have not yet been presented in the literature for decapods.     

Population structure of Macrobrachium australiense (Decapoda: Palaemonidae) in Western Queensland, Australia: the role of contemporary and historical processes

Rivers of Western Queensland, Australia, represent a discontinuous and variable aquatic habitat for the freshwater fauna of the region. Rivers periodically fluctuate between being highly fragmented, with numerous disconnected waterholes and ephemeral channels, and being highly connected by a dominant system of anastomosing channels. We used mitochondrial sequences to investigate the genetic structure and inferred patterns of dispersal associated with this flow regime for the freshwater prawn, Macrobrachium australiense (Decapoda: Palaemonidae), sampling 28 localities throughout eight catchments. Based on a 505 base pair fragment of mitochondrial cytochrome c oxidase subunit I, we identified 98 haplotypes in a sample of 402 individuals. The haplotypes clustered into two main clades corresponding geographically to the major drainages: the Lake Eyre and Murray-Darling basins. Populations of M. australiense inhabiting the two basins appear to have diverged around 800,000 years ago (estimated sequence divergence of 1.6%). Analysis of population differentiation indicated contemporary high levels of genetic subdivision and restricted gene flow among populations within and among catchments. Phylogenetic analysis detected a series of historical range expansions in the region and we suggest that climate fluctuations during the Pleistocene have resulted in extensive floods that have promoted historical movements of aquatic organisms across catchment boundaries.     

Embryo lipoproteins and yolk lipovitellin consumption during embryogenesis in Macrobrachium borellii (Crustacea: Palaemonidae)

The prawn Macrobrachium borellii has lecithotrophic eggs with highly-abbreviated development. The major yolk component is lipovitellin (LV), a lipoprotein with 30% lipids (by weight). LV consumption during embryogenesis was followed by ELISA and Western blot analysis using an anti-LV polyclonal antibody. No cross-reacting proteins were observed and LV-like lipoproteins were strongly recognized by the antibody in hemolymph (vitellogenin), yolk (LV) and embryos (LVe), as determined by Western Blot analysis. LV decreased significantly along development from 9.4 to 1.1 microg/mg egg. Consumption rate of LV was slow in early embryogenesis, followed by a rapid utilization in late embryonic stages. Significant LVe amounts were still present at hatching. LV apolipoproteins were selectively degraded during embryo development, being the highest molecular weight subunit the most affected. Comparison among in vitro, in vivo and theoretical proteolysis suggested that trypsin may be involved in LV degradation during late embryogenesis. Embryo lipoprotein (HDLe) synthesis was first detected at stage 6. HDLe shared the same density, MW and subunit composition as adult hemolymph HDL(1) and did not cross-react with LV-like lipoproteins. Though expressed at low concentration, it fulfilled embryo needs for lipid transport among organs.