First worldwide molecular phylogeny of the morphologically and ecologically hyperdiversified snapping shrimp genus Alpheus (Malacostraca: Decapoda).

Hyperdiverse animal groups raise intriguing questions regarding the factors that generate and maintain their diversity. The snapping shrimp genus Alpheus (with >300 described species) is a spectacularly diversified group of decapod crustaceans that serves as an exemplary system for addressing evolutionary questions regarding morphological adaptations, symbiosis, cryptic diversity and molecular divergence. A lack of information regarding evolutionary relationships among species has limited investigations into the mechanisms that drive the diversification of Alpheus. Previous phylogenetic studies of Alpheus have been restricted in scope, while molecular datasets used for phylogenetic reconstructions have been based solely on mitochondrial and a handful of nuclear markers. Here we use an anchored hybrid enrichment (AHE) approach to resolve phylogenetic relationships among species of Alpheus. The AHE method generated sequence data for 240 loci (>72,000 bp) for 65 terminal species that span the geographic, ecological and taxonomic diversity of Alpheus. Our resulting, well-supported phylogeny demonstrates a lack of monophyly for five out of seven morphologically defined species groups that have traditionally been used as a framework in Alpheus taxonomy. Our results also suggest that symbiotic associations with a variety of other animals have evolved independently in at least seven lineages in this genus. Our AHE phylogeny represents the most comprehensive phylogenetic treatment of Alpheus to date and will provide a useful evolutionary framework to further investigate questions, such as various modifications of the snapping claw and the role of habitat specialization and symbiosis in promoting speciation. Running head: PHYLOGENY OF THE SNAPPING SHRIMP GENUS ALPHEUS.

Morphology and DNA Data Reveal a New Shrimp Species of Genus Latreutes Stimpson, 1860 (Decapoda: Hippolytidae) from the Western Atlantic.

The genus Latreutes Stimpson, 1860 comprises 17 described species of marine shrimps, most of them distributed in the Indo-West Pacific and eastern Pacific. Only three species are recorded in the Atlantic. When comparing specimens of Latreutes parvulus (Stimpson, 1871) from both sides of the Atlantic by means of a combination of morphological and molecular data, we recognized a new species of Latreutes from Brazil (Rio Grande do Norte, Bahia, Rio de Janeiro and São Paulo). The phylogenetic tree generated on the basis of the 16S gene showed a clear separation of the specimens of the new species from the other closely related species analyzed. The analyses also evidenced that the new species and L. parvulus are sister taxa. Although very similar, both species can be easily separated by features of the scaphocerite and eyestalk. Rostrum form and dentition, characters usually used for L. parvulus recognition, were highly variable in both species. Thus, herein the new species is described, illustrated, and compared with morphological similar species of Latreutes. A key for the Atlantic species is also provided.

Ontogenetic shape changes and sexual dimorphism in Aegla marginata Bond-Buckup and Buckup, 1994.

A study on relative growth, sexual dimorphism and ontogenetic trajectory was carried out in a population of the aeglidAegla marginata coming from Barrinha River, Iguape River Basin, Tunas do Paraná, Paraná State, Brazil. The size the of morphological sexual maturity was estimated for males and females. The analysis of sexual dimorphism and ontogenetic trajectory were performed using geometric morphometric technique. Males reach maturity with 10.58 mm of carapace length (CL) and females with 10.38 mm CL. Sexual size dimorphism was only visible among adults, with males reaching larger sizes. This is probably related to the reproductive strategy of males. However, sexual shape dimorphism was found for both juveniles and adults: the posterior region of the carapace was wider in females. As the contrast of this feature was stronger in adults, it can be considered that large abdomen is advantageous for egg incubation. The allometric trajectories of juveniles presented similar directions, becoming divergent during the adult phase. The shape variation inA. marginataoccurred gradually throughout its development, with no abrupt transformation upon reaching sexual maturity. The reproductive adaptation is the main reason for the morphological variation within populations ofA. marginata.

Evolution of body size, vision, and biodiversity of coral-associated organisms: evidence from fossil crustaceans in cold-water coral and tropical coral ecosystems.

BACKGROUND: Modern cold-water coral and tropical coral environments harbor a highly diverse and ecologically important macrofauna of crustaceans that face elevated extinction risks due to reef decline. The effect of environmental conditions acting on decapod crustaceans comparing these two habitats is poorly understood today and in deep time. Here, we compare the biodiversity, eye socket height as a proxy for eye size, and body size of decapods in fossil cold-water and tropical reefs that formed prior to human disturbance. RESULTS: We show that decapod biodiversity is higher in fossil tropical reefs from The Netherlands, Italy, and Spain compared to that of the exceptionally well-preserved Paleocene (Danian) cold-water reef/mound ecosystem from Faxe (Denmark), where decapod diversity is highest in a more heterogeneous, mixed bryozoan-coral habitat instead of in coral and bryozoan-dominated facies. The relatively low diversity at Faxe was not influenced substantially by the preceding Cretaceous/Paleogene extinction event that is not apparent in the standing diversity of decapods in our analyses, or by sampling, preservation, and/or a latitudinal diversity gradient. Instead, the lower availability of food and fewer hiding places for decapods may explain this low diversity. Furthermore, decapods from Faxe are larger than those from tropical waters for half of the comparisons, which may be caused by a lower number of predators, the delayed maturity, and the increased life span of crustaceans in deeper, colder waters. Finally, deep-water specimens of the benthic crab Caloxanthus from Faxe exhibit a larger eye socket size compared to congeneric specimens from tropical reefs, suggesting that dim light conditions favored the evolution of relatively large eyes. CONCLUSIONS: The results suggest a strong habitat control on the biodiversity of crustaceans in coral-associated environments and that the diversity difference between deep, cold-water reefs and tropical reefs evolved at least ~63 million years ago. Futhermore, body size and vision in crustaceans evolved in response to environmental conditions in the deep sea. We highlight the usefulness of ancient reefs to study organismal evolution and ecology.

Intuitive representation of photopolarimetric data using the polarization ellipse.

Photopolarimetry is the spatial characterization of light polarization. Unlike intensity or wavelength, we are largely insensitive to polarization and therefore find it hard to explore the multidimensional data that photopolarimetry produces (two spatial dimensions plus four polarization dimensions). Many different ways for presenting and exploring this modality of light have been suggested. Most of these ignore circular polarization, include multiple image panes that make correlating structure with polarization difficult, and obscure the main trends with overly detailed information and often misleading colour maps. Here, we suggest a novel way for presenting the main results from photopolarimetric analyses. By superimposing a grid of polarization ellipses onto the RGB image, the full polarization state of each cell is intuitively conveyed to the reader. This method presents linear and circular polarization as well as ellipticity in a graphical manner, does not require multiple panes, facilitates the correlation between structure and polarization, and requires the addition of only three novel colours. We demonstrate its usefulness in a biological context where we believe it would be most relevant.

The comparative hydrodynamics of rapid rotation by predatory appendages.

Countless aquatic animals rotate appendages through the water, yet fluid forces are typically modeled with translational motion. To elucidate the hydrodynamics of rotation, we analyzed the raptorial appendages of mantis shrimp (Stomatopoda) using a combination of flume experiments, mathematical modeling and phylogenetic comparative analyses. We found that computationally efficient blade-element models offered an accurate first-order approximation of drag, when compared with a more elaborate computational fluid-dynamic model. Taking advantage of this efficiency, we compared the hydrodynamics of the raptorial appendage in different species, including a newly measured spearing species, Coronis scolopendra The ultrafast appendages of a smasher species (Odontodactylus scyllarus) were an order of magnitude smaller, yet experienced values of drag-induced torque similar to those of a spearing species (Lysiosquillina maculata). The dactyl, a stabbing segment that can be opened at the distal end of the appendage, generated substantial additional drag in the smasher, but not in the spearer, which uses the segment to capture evasive prey. Phylogenetic comparative analyses revealed that larger mantis shrimp species strike more slowly, regardless of whether they smash or spear their prey. In summary, drag was minimally affected by shape, whereas size, speed and dactyl orientation dominated and differentiated the hydrodynamic forces across species and sizes. This study demonstrates the utility of simple mathematical modeling for comparative analyses and illustrates the multi-faceted consequences of drag during the evolutionary diversification of rotating appendages.

Size-dependent mortality rate profiles.

Knowledge of mortality rates is crucial to the understanding of population dynamics in populations of free-living fish and invertebrates in marine and freshwater environments, and consequently to sustainable resource management. There is a well developed theory of population dynamics based on age distributions that allow direct estimation of mortality rates. However, for most cases the aging of individuals is difficult or age distributions are not available for other reasons. The body size distribution is a widely available alternative although the theory underlying the formation of its shape is more complicated than in the case of age distributions. A solid theory of the time evolution of a population structured by any physiological variable has been developed in 1960s and 1970s by adapting the Hamilton-Jacobi formulation of classical mechanics, and equations to estimate the body size-distributed mortality profile have been derived for simple cases. Here I extend those results with regards to the size-distributed mortality profile to complex cases of non-stationary populations, individuals growing according to a generalised growth model and seasonally patterned recruitment pulses. I apply resulting methods to two cases in the marine environment, a benthic crustacean population that was growing during the period of observation and whose individuals grow with negative acceleration, and a sea urchin coastal population that is undergoing a stable cycle of two equilibrium points in population size whose individuals grow with varying acceleration that switches sign along the size range. The extension is very general and substantially widens the applicability of the theory.

[Morfogeometry shell crab Damithrax spinosissimus (Decapoda: Mithracidae) on three islands in the southwestern Caribbean]. / Morfogeometría del caparazón del cangrejo Damithrax spinosissimus (Decapoda: Mithracidae) en tres islas del Caribe suroccidental.

The Caribbean King Crab, Damithrax spinosissimus is a fishery resource, but few biological studies are available. Its patchy distribution, and the high environmental heterogeneity due to the oceanographic, landscape, and ecological characteristics of the Caribbean Sea, can favor the phenotypic variability according to the geographic origin. For this reason, the objective of our study was to determine morphometic variability in the carapace of the crabs from three Southwestern Caribbean islands: Providence, Rosario and San Bernardo. The former has an oceanic influence, whereas the two latter islands, which are closer to the mainland, have more influence of the coastal dynamic. A total of 276 individuals from the three islands were captured and photographed: 103 females and 173 males; their variation was analyzed from 12 anatomical landmarks marked on one half of the carapace. The differences in the carapace size were calculated using a Fisher's pairwise comparison; the allometric effect was calculated by Multivariate Regression Analysis; and the Allometric model via Multivariate Analysis of Covariance. The average shape was calculated from the relative wraps RW obtained through PCA analysis; and the Euclidian distances between the shape averages, were used to construct a tree using the Neighbour-Joining algorithm over 10 000 permutations. The results showed significant differences in the size and shape of the carapace between sexes and among the three islands. The differences in the shape of the crabs from Providencia were significantly greater than those found between the crabs of Rosario and San Bernardo. These results can be explained inclusively due to the genetic differences and phenotypic plasticity, due to environmental heterogeneity of the sector. This study, the first of its kind, is a contribution to the knowledge of the species. We concluded that different management strategies should be differently adopted in each of the three studied islands.

[Evaluation of the growth of Callinectes sapidus (Decapoda: Portunidae) by the use of lengthbased methods based on size in Tamaulipas, Mexico]. / Evaluación del crecimiento de Callinectes sapidus (Decapoda: Portunidae) con métodos basados en talla, Tamaulipas, México.

The capture blue crab (Callinectes sapidus) is one of the major fisheries of the state of Tamaulipas, Mexico; both in volume and selling price, as well as employment generation, but there is little information on its biological characteristics. The aim of this study was to evaluate the growth parameters of the blue crab, establishing the most appropriate method. We estimated the length frequency of 17 814 crabs from commercial catch of thirteen locations, including four coastal lagoons. The lagoons were El Barril, Madre, Morales and San Andrés from Tamaulipas, State. Growth parameters were evaluated using indirect methods ELEFAN, PROJMAT and SLCA in combination with the jackknife technique to establish the uncertainty of estimates inherent in each method. The growth parameters L∞ and k were consolidated for purposes of comparison with the growth index phi prime (Φ'). With a mode of 110 mm, the interval carapace length varied between 60 and 205 mm. The values of the growth parameters varied according to the method used. Using SLCA, L∞ varied between 259 and 260 mm and k ranged between 0.749 and 0.750 /year; with PROJMAT, L∞ recorded values between 205 and 260 mm, k fluctuated between 0.550 and 0.740/year, and with ELEFAN, L∞ ranged between 156 and 215 mm and k varied between 0.479 and 0.848/year. Estimates by jackknife detected no variability in Φ' between locations and significant differences between methods. The ranges of values of Φ' and PROJMAT estimated SLCA (4.70 to 4.71 and 4.66 to 4.70, respectively) were in the range reported in the literature (4.201-4.798), while lower values ELEFAN contributed significantly (3.87 to 4.27). The SLCA and PROJMAT methods in combination with the jackknife technique, proved to be the most suitable to estimate the growth parameters of C. sapidus.

Social Control of Reproduction and Breeding Monopolization in the Eusocial Snapping Shrimp Synalpheus elizabethae.

Understanding why individuals within altruistic societies forgo reproduction to raise others' offspring has fascinated scientists since Darwin. Although worker polymorphism is thought to have evolved only in sterile workers, worker subcastes appear to be common among social invertebrates and vertebrates. We asked whether sterility accompanies eusociality and morphological differentiation in snapping shrimps (Synalpheus)-the only known marine eusocial group. We show that workers in Synalpheus elizabethae are reproductively totipotent and that female-but not male-gonadal development and mating are mediated by the presence of a queen, apparently without physical aggression. In queenless experimental colonies, a single immature female worker typically became ovigerous, and no female workers matured in colonies with a resident queen. Thus, eusocial shrimp workers retain reproductive totipotency despite signs of morphological specialization. The failure of most female workers to mature is instead facultative and mediated by the presence of the queen, ensuring her reproductive monopoly.

Large-field electron imaging and X-ray elemental mapping unveil the morphology, structure, and fractal features of a Cretaceous fossil at the centimeter scale.

We used here a scanning electron microscopy approach that detected backscattered electrons (BSEs) and X-rays (from ionization processes) along a large-field (LF) scan, applied on a Cretaceous fossil of a shrimp (area ∼280 mm(2)) from the Araripe Sedimentary Basin. High-definition LF images from BSEs and X-rays were essentially generated by assembling thousands of magnified images that covered the whole area of the fossil, thus unveiling morphological and compositional aspects at length scales from micrometers to centimeters. Morphological features of the shrimp such as pleopods, pereopods, and antennae located at near-surface layers (undetected by photography techniques) were unveiled in detail by LF BSE images and in calcium and phosphorus elemental maps (mineralized as hydroxyapatite). LF elemental maps for zinc and sulfur indicated a rare fossilization event observed for the first time in fossils from the Araripe Sedimentary Basin: the mineralization of zinc sulfide interfacing to hydroxyapatite in the fossil. Finally, a dimensional analysis of the phosphorus map led to an important finding: the existence of a fractal characteristic (D = 1.63) for the hydroxyapatite-matrix interface, a result of physical-geological events occurring with spatial scale invariance on the specimen, over millions of years.

The emergence of lobsters: phylogenetic relationships, morphological evolution and divergence time comparisons of an ancient group (decapoda: achelata, astacidea, glypheidea, polychelida).

Lobsters are a ubiquitous and economically important group of decapod crustaceans that include the infraorders Polychelida, Glypheidea, Astacidea and Achelata. They include familiar forms such as the spiny, slipper, clawed lobsters and crayfish and unfamiliar forms such as the deep-sea and "living fossil" species. The high degree of morphological diversity among these infraorders has led to a dynamic classification and conflicting hypotheses of evolutionary relationships. In this study, we estimated phylogenetic relationships among the major groups of all lobster families and 94% of the genera using six genes (mitochondrial and nuclear) and 195 morphological characters across 173 species of lobsters for the most comprehensive sampling to date. Lobsters were recovered as a non-monophyletic assemblage in the combined (molecular + morphology) analysis. All families were monophyletic, with the exception of Cambaridae, and 7 of 79 genera were recovered as poly- or paraphyletic. A rich fossil history coupled with dense taxon coverage allowed us to estimate and compare divergence times and origins of major lineages using two drastically different approaches. Age priors were constructed and/or included based on fossil age information or fossil discovery, age, and extant species count data. Results from the two approaches were largely congruent across deep to shallow taxonomic divergences across major lineages. The origin of the first lobster-like decapod (Polychelida) was estimated in the Devonian (∼409-372 Ma) with all infraorders present in the Carboniferous (∼353-318 Ma). Fossil calibration subsampling studies examined the influence of sampling density (number of fossils) and placement (deep, middle, and shallow) on divergence time estimates. Results from our study suggest including at least 1 fossil per 10 operational taxonomic units (OTUs) in divergence dating analyses. [Dating; decapods; divergence; lobsters; molecular; morphology; phylogenetics.].

Additional records of the laomediid mud-shrimp genus Naushonia Kingsley, 1897 (Crustacea: Decapoda: Gebiidea), with a revised identification key.

Six species of the peculiar mud-shrimp genus Naushonia Kingsley, 1897 (Laomediidae) from various localities in the world are treated in this study: N. carinata Dworschak, Marin & Anker, 2006 (newly recorded from Japan); N. japonica Komai, 2004 (second record since the original description); N. lactoalbida Berggren, 1992 (newly recorded from Papua New Guinea); N. portoricensis (Rathbun, 1901) (newly recorded from the French Antilles); N. serratipalma Komai & Anker, 2010 (newly recorded from Saudi Arabia); and N. draconis Anker, 2014 (newly recorded from Cuba). A detailed description and new illustrations are provided for N. portoricensis, a species for which a modern description was hitherto unavailable. Gill formula was re-examined in all six species. Colour photographs of living or fresh specimens are provided for most species. A revised key to all known species of Naushonia is presented, a eliminating misinterpretation of an important character in previous keys.

Further records of species of Gennadas (Crustacea, Decapoda, Dendrobranchiata, Benthesicymidae) in the Mexican Pacific.

Additional material of species of Gennadas was collected off western Mexico during the TALUD survey aboard the R/V "El Puma". Gennadas sordidus was by far the most common species collected and only a few specimens of the other three species, G. incertus, G. propinquus and G. scutatus, were obtained. New and previous records indicate that Gennadas sordidus is widely distributed along the west coast of Mexico, although there is only one record for it south of the Gulf of California. In this study it was collected in as many as 35 localities, both in the Gulf of California and off the west coast of Southern Baja California. Numbers of specimens also confirmed that it is the dominant species of Gennadas in the area. Gennadas incertus was found in six samples, all from off the west coast of northern Baja California. The other two species occurred each in two samples only, with sampling localities widely spread along the Baja California Peninsula. Compared with the rest of the eastern Pacific, the diversity of Gennadas in Mexican waters is intermediate (6 species) vs. 9 in Chile and 4 in the NW Pacific.

Description of the newly-hatched juvenile of Aegla perobae (Crustacea: Decapoda: Aeglidae).

The present paper contains the complete description of the external morphology of the first juvenile stage of Aegla perobae analysed through light microscopy (LM) and scanning electron microscopy (SEM). Newly-hatched juveniles were obtained from ovigerous females kept under laboratory conditions. Hatching is asynchronous, taking 2-4 days for all juveniles of a single brood to hatch. Average carapace dimensions are 1.54 mm wide and 1.69 mm long (rostrum excluded). Morphology of the carapace, of the cephalothoracic appendages (antennule, antenna, mandible, maxillule, maxilla, maxillipeds, and pereopods), of the pleon, and of the tail fan (telson plus uropods) are described in detail. Aegla perobae juveniles can be readily differentiate from the first juveniles of other aeglids species described so far by the upwardly curved condition of the distal region of the rostrum and the distinct groove along the orbital sinus produced the elevated free in this area. Pleopods 2-5 are present as rudimentary digitiform buds. Rudimentary pleopods are still present in adult males of the species, a trait not yet described in freshwater aeglids. This curious condition is compared and discussed in the light of the current knowledge of early postembryonic developmental patterns found in other anomurans.

A new species of the ghost shrimp genus Lepidophthalmus (Crustacea: Decapoda: Axiidea) from the southwestern Gulf of Mexico.

A new species of Lepidophthalmus lacking a ventral median sclerite on the second abdominal somite is described from coastal waters of the southwestern Gulf of Mexico. Lepidophthalmus statoni sp. nov., originally recognized only as a unique population in allozyme studies, is sympatric with the ventrally plated species Lepidophthalmus manningi Felder & Staton, 2000, but more closely resembles Lepidophthalmus louisianensis (Schmitt, 1935) from the northern and northwestern Gulf of Mexico. Apparently restricted to intertidal and shallow subtidal tropical waters, the new species is known to range from western Campeche to middle-upper reaches of Veracruz, Mexico. As many members of the genus, it commonly inhabits euryhaline inlets, estuaries, and protected shorelines, including richly organic muddy to clayey sands and sandy muds adjacent to shoreline vegetation. Coloration is documented and discussed as a tool to facilitate field identifications, as are morphological characters.

Alpheopsis balaeniceps sp. nov., an unusual alpheid shrimp (Decapoda: Caridea) from French Polynesia.

Alpheopsis balaeniceps sp. nov. is described based on a single male specimen from Moorea, French Polynesia. The new species is unique within the genus Alpheopsis Coutière, 1896 in possessing balaeniceps-type setae on the fingers of both chelipeds, being especially well developed on the major chela. Within the family Alpheidae, this feature exists only in some species of the distantly related genus Alpheus Fabricius, 1798, however, only on the minor cheliped. This convergence between Alpheopsis and Alpheus is another example of the remarkable diversity and plasticity of the alpheid chelipeds.

Extending the southern range of four shrimps (Crustacea: Decapoda: Stenopodidae, Hippolytidae and Alpheidae) in southwestern Atlantic (27o S) and confirming the presence of Mediterranean Stenopus spinosus Risso, 1827 in Brazil.

In subtidal zones, certain shrimp species with cryptic behaviour represent a gap in the biodiversity description in many places in the world. This study extends the southern limit of Stenopus hispidus (Oliver, 1811), Alpheus formosus Gibbes, 1850, Alpheus cf. packardii Kingsley, 1880 and Lysmata ankeri Rhyne & Lin, 2006 to Santa Catarina State-Brazil, 27oS. The results also confirm the new occurrence of Stenopus spinosus Risso, 1827 in Brazilian waters. All specimens were collected by scuba diving from rocky islands between 3 and 25 meters depth. We present for each species certain taxonomic features in colour images that will help to identify these decapods in situ in further monitoring programs.

Phylogenetic relationships within the snapping shrimp genus Synalpheus (Decapoda: Alpheidae).

The snapping shrimp genus Synalpheus (Alpheidae) is one of the most speciose decapod genera, with over 160 described species worldwide. Most species live in symbiotic relationships with other marine organisms, such as sponges, corals and crinoids, and some sponge-dwelling species have a highly organized, social structure. The present study is the first worldwide molecular phylogenetic analysis of Synalpheus, based on >2,200 bp of sequence data from two mitochondrial (COI and 16S) and two nuclear (PEPCK and 18S) loci. Our molecular data show strong support for monophyly of three out of six traditionally recognized morphology-based species groups: the S. brevicarpus, S. comatularum and S. gambarelloides groups. The remaining three species groups (S. paulsoni, S. neomeris and S. coutierei groups) are non-monophyletic in their current composition and will need to be either abandoned or taxonomically redefined. We also identified potential cryptic species of Synalpheus in our dataset, using intraspecific and interspecific sequence variation in COI from the taxonomically well-studied S. gambarelloides group to establish a genetic divergence threshold. We then used both genetic divergence and tree-based criteria (reciprocal monophyly) to identify potential cryptic species in the remaining taxa of the genus. Our results suggest the presence of multiple cryptic lineages in Synalpheus, underlining the need for more integrative taxonomic studies-including morphological, ecological, molecular, and color pattern data-in this biologically interesting genus.

New records of the caridean shrimp genus Processa Leach, 1815 (Crustacea: Decapoda: Processidae) from Japan, with descriptions of two new species.

Three species of the processid shrimp genus Processa Leach, 1815, including two new species, are reported from shallow coastal waters in Okinawa Island, Ryukyu Islands, Japan: P. affinis Hayashi, 1975, P. filipes n. sp. and P. hayashii n. sp. The present specimens of P. affinis represent the rediscovery and range extension of this poorly known species, originally described from Indonesia. Male characteristics of P. affinis are documented for the first time. The two new species are referred to the P. aequimana Paulson, 1875 species group. Processa filipes n. sp. is characteristic in the greatly elongate fifth pereopod and the transverse ridge on thoracic sternite 8 consisting of soft cuticle. Processa hayashii n. sp. is characterized by the presence of a deep concavity on the anterolateral margin of the carapace just inferior to the antennal tooth. The number of species of Processa known from Japanese waters is raised from five to eight.