Feeding activity of Callinectes ornatus Ordway, 1863 and Callinectes danae Smith, 1869 (Crustacea, Brachyura, Portunidae) in Ubatuba, SP, Brazil

The feeding activity along the day cycle and the time consumed for extracellular digestion were evaluated in the portunids C. ornatus and C. danae. Swimming crabs were obtained from trawling in Ubatuba bay, São Paulo, Brazil, during both the rainy and dry seasons. In each season, daily scheduled samples were taken at dawn (±6 h), noon (±12 h), dusk (±18 h) and midnight (±24 h). All individuals were dissected and the degree of stomach replenishment was recorded. In order to estimate the time elapsed for extracellular digestion, crabs were fed, and groups were dissected at 30 min intervals to check the conditions of their stomachs. In general, both species show a higher feeding activity during periods of lower light intensity, as evidenced by an increased percentage of full stomachs in dusk and midnight samples. The obtained results support higher feeding activity at night in these species and indicate short time for extracellular digestion, not exceeding 8 h. Nevertheless, full stomachs were recorded in all sampling schedules. In this case, it should be considered that elimination of certain food items such as fish bones, mollusk shells and carapace fragments of crustaceans could take more time than other items. Additionally, some crab species could require a cycle of cell replacement in the midgut gland epithelium until they can take their next meal.     

Female reproductive pattern of Callinectes sapidus Rathbun, 1896 (Brachyura: Portunidae) in Iskenderun Bay,Eastern Mediterranean

We studied the female reproductive pattern of Callinectes sapidus, which was introduced to the Mediterranean in the 20^th century. We assessed female size at first maturity, fecundity, and fecundity relationship to size in Iskenderun Bay, eastern Mediterranean, Turkey. Samples were collected between July 2014 and June 2015 using bottom trawling at depths ranging from 1 to 50 m. A total of 322 crabs were caught of which 308 (95.7%) were females including 116 ovigerous ones. The minimum carapace width of the mature females was 39.1 mm and the mean carapace width 123.8 mm. The carapace width of ovigerous females varied between 95.1 and 144.5 mm, with a mean of 120.3 mm. The highest number of ovigerous females was observed in July and August. Mean fecundity was 1.91 million (667,950–4,669,853) eggs per female. A weak positive linear relationship between fecundity and carapace width was noted, as well as a high correlation with total egg weight. In the eastern Mediterranean, maturity sizes of females were smaller than those in the native region of the species.     

Identification of polymorphic microsatellite loci in the mud crab Scylla serrata (Brachyura: Portunidae)

Five microsatellite loci are identified and characterized from the genome of Scylla serrata, a widespread and commercially important species of coastal marine crab. The loci were detected by randomly screening for di‐ and tri‐nucleotide repeat units within a partial genomic library developed for the species. The five loci consist of dinucleotide repeats and are both co‐dominant and polymorphic within the species. A sample (N = 36) of S. serrata from one Australian population has an average observed heterozygosity of 0.875 and provides no evidence of either linkage among loci or significant deviation from random mating expectations across loci. PCR products for each of the five loci were also observed from a small sample of three other species within the Scylla genus. These markers may provide genetic information that will be useful for both aquaculture and studies of natural populations of the genus.     

Morphology of the male reproductive system and sperm ultrastructure of the egg parasitoid Gryon pennsylvanicum (Ashmead) (Hymenoptera,Platygastridae)

Gryon pennsylvanicum is a platygastrid hymenopteran that has lately received increasing attention in Europe due to its possible use in biological control of the conifer seed bug pest Leptoglossus occidentalis. Here the male reproductive system and the spermatogenesis of this species, along with those of Gryon muscaeformis, are examined ultrastructurally for the first time. The male genital system is formed by a pair of testes, each containing only one follicle, a pair of accessory glands and deferent ducts connected to a single ejaculatory duct. All the stages of spermatogenesis are described in detail. Characteristic features of the Gryon spp. sperm, which are 100 μm long, are the presence of a polygonal nucleus, only one mitochondrial derivative, the occurrence of the centriole adjunct and a typical insect 9 + 9 + 2 flagellar axoneme. The single derivative, however, results from a process in which one of the two mitochondria is lost during spermiogenesis. Unlike in other insects, two centrioles occur in spermatids as a consequence of the ameiotic parthenogenesis. These characteristics stand as a valuable tool for phylogenetic inferences. Furthermore this study suggests a useful strategy for laboratory mass rearing.     

Molluscan predation by Ovalipes punctatus (De Haan) (Crustacea: Brachyura: Portunidae)

Molluscan predation by the three-spot swimming crab was investigated. The dentition of the heteromorphic chelae allowed crushing, shearing, cutting and holding of prey. Laboratory investigations indicated that small mussels and gastropods were crushed, the larger mussels were prized open, and the foot of the larger gastropods shredded and bits removed. Stomach contents of freshly captured crabs indicated that the crabs are selective carnivores and preferred prey species which are not most abundant in situ (crabs from Kings Beach, Donax serra Röding; crabs from Maitlands River Beach, Bullia rhodostoma Reeve). Ovalipes punctatus (De Haan) foraged on a variety of prey and had no upper prey size limit, but the crabs did show preferences for certain prey sizes. Data indicate that the swimming crabs can effectively utilize the entire mollusc populations on the beaches as prey items.     

Morphology and development of the male reproductive tract in Callinectes danae (Crustacea: Brachyura)

The aim of this study was to characterize the morphology and function of each section of the reproductive system of male Callinectes danae, as well as the stages of reproductive development and their relation to secondary sexual characteristics. Development of their reproductive system begins after completion of the pubertal moult. The growth of the gonopodium showed negative allometry for both juveniles and adults. The reproductive system is divided into portions with different functions. There is a germinal zone in the testes containing spermatogonia, a zone of maturation containing spermatocytes, spermatids or spermatozoa, as well as a collecting duct, which carries spermatozoa to the vas deferens. There are two matrices in the anterior vas deferens that initiate the separation of spermatozoa groups, one composed of polysaccharide acids (matrix I) and another consisting of neutral polysaccharides (matrix II). In the median vas deferens, the matrix II forms an acellular capsule, which forms the spermatophores. In the posterior vas deferens, the matrices are accumulated, initially with a granular texture and are homogenous for the final portion. The ejaculatory duct and penis have muscle lining to expel the spermatophores at copulation. Even after copulation, males retain a stock of spermatophores, allowing copulation with other females.     

The ovarian cycle histochemistry and its relationship with hepatopancreas weight in the blue crab Callinectes danae (Crustacea: Portunidae)

Zara, F.J., Gaeta, H.H., Costa, T.M., Toyama, M.H. and Caetano, F.H. 2011. The ovarian cycle histochemistry and its relationship with hepatopancreas weight in the blue crab Callinectes danae (Crustacea: Portunidae). —Acta Zoologica (Stockholm) 00 :1–13. Several studies use macroscopic patterns of the ovarian development in crustaceans. Here, we examined the relationship between ovary histochemistry, changes in gonadosomatic and hepatosomatic indices against the macroscopic pattern of the ovarian development in Callinectes danae. Animals were collected in the south coast of São Paulo State, Brazil. Ovaries were macroscopically classified as juvenile, rudimentary, developing, intermediary, mature, and rudimentary ovigerous. Samples were fixed in 4% paraformaldehyde, processed for historesin, and stained with HE, protein, and neutral and acid polysaccharides detection. The juvenile oocytes are not enclosed by follicular cells and have fewer yolk nuclei being less intense in PAS reactivity than rudimentary oocytes. Developing oocytes show yolk granules and thick follicular cells. Yolk granules were positive for proteins and neutral polysaccharides. The intermediary stage is marked by a qualitative increase in yolk granules and the onset of chorion formation. In mature oocytes, cytoplasm is completely filled by yolk granules and the chorion is completely formed. Ovigerous ovaries have several atretic follicles and large quantities of hemocytes in the process of tissue reorganization. In C. danae, the changes in cell, goandosomatic and hepatosomatic indices coinciding with macroscopic observations and any combination of different macroscopic stages in a single pattern should be avoided.     

Comparative spermatozoal ultrastructure and molecular analysis in dromiid crabs and their phylogenetic implications for Dromiidae and Podotremata (Decapoda: Brachyura)

We described the spermatozoal ultrastructure and conducted a molecular analysis of Dromiidae Hypoconcha parasitica, Hypoconcha arcuata, Moreiradromia antillensis and Dromia erythropus. To elucidate the relationship between the different species of this brachyuran group, we also compared the spermatozoal morphologies and phylogenetic positioning among species of Dromiidae, Dromioidea and Podotremata. Specimens were collected from the northern coast of São Paulo, Brazil and were fixed and processed followed by transmission electron microscopy and molecular analysis routines. The Dromiidae spermatozoa studied are characterized by a discoidal acrosome, with three or four concentric zones, which are centrally separated by a bilaterally capitate perforatorial chamber, with a “mushroom”-shaped apex in the Hypoconchinae and a “T-shape” in Dromiinae. Above the perforatorial chamber, there is an apical protuberance, continuous with the subopercular region and the operculum, which forms a low, centrally perforated dome. Under differential interference contrast microscopy, the spermatozoa show 3 to 4 radial arms. The spermatozoal characters in Hypoconchinae and Dromiinae do not separate these subfamilies from the Dromiidae and Dromioidea. Ultrastructural differentiation was only found between representative Dromioidea and other Podotremata. Thus, the spermiotaxonomy of these Hypoconcha, Moreiradromia and Dromia species corroborated previous morphological and molecular studies, supporting the monophyly of Dromiidae and Dynomenidae in relation to Homolidae and Latreilliidae.     

Ultrastructure of spermatozoa and spermatophores of old world freshwater crabs (Brachyura: Potamoidea: Gecarcinucidae,Potamidae, and Potamonautidae)

We investigated the ultrastructure of spermatozoa and spermatophores of 19 palaeotropical freshwater crab species [12 species of the Gecarcinucidae, 6 of the Potamidae (Potamiscinae), and 1 species of the Potamonautidae (Deckeniinae: Hydrothelphusini)]. The investigated Potamiscinae have densely packed coenospermic spermatophores with the exception of Thaiphusa sirikit and Johora singaporensis that exhibit cleistospermia. In contrast, in the Gecarcinucidae the spermatozoa are loosely embedded in a mucous matrix. The gecarcinucid and potamiscine sperm differ, furthermore, in acrosomal structure and size. The acrosome in the Gecarcinucidae is much smaller and spherical, while the larger acrosome in the Potamiscinae has the tendency to be depressed. In the Potamiscinae, an additional middle acrosomal zone evolved between the acrosome ray zone and the outer acrosomal zone. Within the Gecarcinucidae, a differentiation into two groups (Gecarcinucinae and Parathelphusinae) is not supported by the present spermatological data. The sperm morphology of Hydrothelphusa aff. madagascariensis (Potamonautidae: Deckeniinae) differs from Potamonautes sidneyi (Potamonautidae: Potamonautinae) in acrosomal size and shape, and in the absence of a periopercular rim. A closer relationship of Deckeniinae and Gecarcinucidae cannot be confirmed by spermatology. J. Morphol., 2009. © 2008 Wiley‐Liss, Inc.     

Molecular phylogeny of the western Atlantic species of the genus Portunus (Crustacea, Brachyura, Portunidae)

The genus Portunus encompasses a comparatively large number of species distributed worldwide in temperate to tropical waters. Although much has been reported about the biology of selected species, taxonomic identification of several species is problematic on the basis of strictly adult morphology. Relationships among species of the genus are also poorly understood, and systematic review of the group is long overdue. Prior to the present study, there had been no comprehensive attempt to resolve taxonomic questions or determine evolutionary relationships within this genus on the basis of molecular genetics. Phylogenetic relationships among 14 putative species of Portunus from the Gulf of Mexico and other waters of the western Atlantic were examined using 16S sequences of the rRNA gene. The resultant molecularly based phylogeny disagrees in several respects with current morphologically based classification of Portunus from this geographical region. Of the 14 species generally recognized, only 12 appear to be valid. We recommend that P. vossi be hereafter regarded as a junior synonym of P. spinimanus and that P. bahamensis be regarded as a junior synonym of P. depressifrons . Our analysis suggests that western Atlantic members of the genus can be subdivided into at least three well-defined clades. Pending further molecular analyses with a large subset of species, it appears that the genus is not monophyletic and that it warrants further taxonomic revision.  © 2007 The Linnean Society of London, Zoological Journal of the Linnean Society , 2007, 150 , 211–220.     

A comparative study of zoeal morphology in the genus Liocarcinus (Crustacea: Brachyura: Portunidae)

The zoeal morphology of Liocarcinus arcuatus, L. corrugatus and L. depurator is described from laboratory reared material and the larvae of L. holsatus, L. puber, L. pusillus and L. marmoreus are reexamined from specimens deposited in the BM (NH). No single meristic character was found that would adequately separate the seven described species of Liocarcinus. Characters that may facilitate separation of later zoeal stages only are listed.     

Geographic and gender shape differences in the carapace of Liocarcinus depurator (Brachyura: Portunidae) using geometric morphometrics and the influence of a digitizing method

The current work describes the use of geometric morphometrics in the study of the portunid crab Liocarcinus depurator carapace shape variation along the Mediterranean coast of Spain. As a first step, the differences in shape induced by digitizing the carapace with either a digital camera or a flat bed scanner were investigated. Carapace shape information inferred from the camera images proved different from that obtained from the scanner images. This difference was greater than any shape variation because of crab gender or sample location. Carapace shape was analysed using multiple analysis of covariance (with centroid size as a covariate) of the partial warps and uniform components indicating differences between genders and between sample location; however, significant three-way interaction implied that such differences were gender and location specific. Landmarks identifying carapace width, carapace length and posterior carapace width showed greatest variation; hence gender and location differences were further analysed through the interrelationships between these linear measures. Regression analysis of the relationship between posterior carapace width and the ratio of carapace width to carapace length indicated clear differences between locations but not between genders. Crabs from the Alborán Sea had proportionally wider posterior carapaces than those from Alicante or North Catalonia. This finding may represent the difference between a southern influx of an Atlantic population distinguishable from a more northerly Mediterranean one, which would be in agreement with the overall oceanographic surface circulation pattern in the western Mediterranean.     

Ultrastructure variation in the spermatozoa of Pseudopaludicola frogs (Amphibia,Anura, Leptodactylidae), with brief comments on its phylogenetic relevance

The taxonomic history of the small frogs of the genus Pseudopaludicola from South America has been controversial. Phylogenetic inferences based on molecular data have identified four Pseudopaludicola clades, correlating with the known variation in karyotypes (2n = 22, 20, 18, and 16). In this study, the ultrastructure of the spermatozoa was analyzed in 12 species of the Pseudopaludicola, with the aim of describing their morphology and identifying characters that may contribute to a better understanding of the phylogenetic relationships. The spermatozoa presented marked differences in tail structures. The tails of the spermatozoa of the species with 2n = 22 chromosomes (Pseudopaludicola sp. 1 [P. pusilla group], Pseudopaludicola falcipes, P. mineira, and Pseudopaludicola saltica), as well as Pseudopaludicola ameghini and Pseudopaludicola ternetzi (2n=20), have juxta‐axonemal fibers, undulating membranes and axial fibers. In contrast, in the species with 2n = 18 (P. facureae, P. giarettai, Pseudopaludicola canga, P. atragula, and Pseudopaludicola sp. 2) and 2n = 16 (Pseudopaludicola mystacalis), there are no evident axial or juxta‐axonemal fibers, but a paraxonemal rod with a thick undulating membrane, which is shorter than that found among Pseudopaludicola species. The ultrastructural morphological differences observed in the spermatozoa of these species may be phylogenetically informative, given that they coincide with the consensus phylogeny of the group and appear to represent a progressive simplification of the spermatozoon. J. Morphol. 276:1495–1504, 2015. © 2015 Wiley Periodicals, Inc.     

Ultrastructure and histochemistry of the metacercarial cyst of Spelotrema nicolli (Microphallidae: Trematoda)

Based on electron microscopy and histochemical tests, four layers are described for the metacercarial cyst of Spelotrema nicolli. The outermost layer I is made up of host connective tissue. Layer II, which may contain lipid, is a narrow band varying in electron density. Layer III is composed of mucoprotein with a granular fine structure. The innermost layer is made up of columnar, proteinaceous crystals, which radiate out from the inner edge of the cyst. Observations on the epidermis of Spelotrema nicolli contrast previous work on the mechanism of cyst formation in microphallids.     

Morphology and ultrastructure of the nephridial system of Hypania invalida (Grube, 1860) (Annelida,Polychaeta, Ampharetidae)

The excretory organs of the freshwater polychaete Hypania invalida have been examined using scanning and transmission electron microscopy. Three pairs of macroscopically and ultrastructurally different nephridia are present in the thorax. Intersegmental septa in the thorax are absent, with the exception of a single diaphragm between second and third chaetiger. The first pair of nephridia is anterior to this septum, the second pair crosses the septum, with the nephrostomes anterior and the ducts and the nephridiopori posterior to it, and the third pair of nephridia is entirely posterior to the diaphragm. The first two pairs of nephridia have ciliated nephrostomes of moderate size and long nephridial ducts that extend the length of the thorax. In contrast, the third pair is characterized by short ducts and very prominent nephrostomes. Macroscopically, seven different sections of nephridial duct cells can be distinguished along the length of the first two pairs of nephridia, whereas, on an ultrastructural basis, only six different regions can be identified. Only two regions of different duct cells can be recognized in the third pair of nephridia. Cells of the two anterior pairs of nephridia show typical characteristics of transport epithelia and most likely function as excretory organs. In contrast, the duct cells of the third pair are not that much differentiated and might primarily be responsible for the release of sexual products, as sperm was observed passing through these ducts. Podocyte‐like cells were observed to accompany nephridial ducts. J. Morphol., 2011. © 2010 Wiley‐Liss, Inc.     

粉尘螨生殖系统超微结构的透射电镜观察

本研究主要采用透射电镜观察粉尘螨Dermatophagoides farinae (Hughes)生殖系统超微结构。粉尘螨雄性生殖系统是由精巢、 输 精管、 附腺、 射精管、 交配器官及附属交配器官组成。精巢内可同时有精子发育各阶段的细胞。精子无核膜、 核染色质聚集成束、 线 粒体缺乏典型的嵴、 胞质内有平行排列的电子致密薄片等为其特征性结构。雌性生殖系统由交合囊、 交合囊管、 储精囊、 囊导管、 卵 巢、 输卵管、 子宫及产卵管构成。卵巢内可见含多个细胞核的中央细胞, 其周为卵母细胞等生殖细胞。该研究丰富了对粉尘螨生殖系统 结构的认识。     

Anatomy and ultrastructure of the male reproductive system in Pleioplana atomata (Platyhelminthes: Polycladida)

Abstract. The ultrastructure of the male reproductive system in the polyclad flatworm Pleioplana atomata is described. Numerous testes are scattered throughout the entire body but are heavily concentrated on the ventral side. All stages of differentiating sperm cells are present in all testes follicles. Intercellular bridges connect spermatocytes and spermatids derived from a single spermatogonium. In the distal part of spermatids, a zone of differentiation develops with a row of microtubules beneath the plasmalemma. Adjacent to these microtubules, an intercentriolar body is flanked by two basal bodies that give rise to two axonemes (each with a 9+“1” microtubular pattern) that face in opposite directions. The Golgi complex appears in the central portion of the spermatid and produces numerous small and large electron-dense bodies. The small bodies surround the nucleus, whereas the large bodies cluster along with the mitochondria in the central part of the spermatid. Development of the spermatid leads to cell elongation and formation of a filiform, biflagellate mature spermatozoon with cortical microtubules all along the sperm shaft. The male canal system consists of paired vasa deferentia that separately enter a single seminal vesicle. A single prostatic canal connects the seminal vesicle to the prostatic vesicle. Ultrastructurally, the seminal vesicle and prostatic canal are very similar, and along with the prostatic vesicle and stylet pocket, are lined by a ciliated epithelium. The ultrastructure of the prostatic vesicle indicates that it probably produces a large volume of seminal fluid that, along with spermatozoa, is transferred to the mating partner through a stylet. Some of the findings, particularly on sperm ultrastructure, may provide characters useful for phylogenetic analysis.     

Structure of the male reproductive system of the blue swimmer crab Portunus pelagicus (Decapoda: Portunidae)

An attempt was made here to study the structure of the male reproductive system of Portunus pelagicus, which would improve the knowledge base on the reproductive biology of the species and also help in the maintenance of broodstock under controlled conditions. Male P. pelagicus of different sizes were collected from the Palk Bay off Mandapam (9°17′ N, 79°9′ E) and maintained under controlled conditions for the study. Tissues from testis, anterior vas deferens (AVD), median vas deferens (MVD), posterior vas deferens (PVD), ejaculatory duct and penis were fixed in Bouin's fluid and 2.5% buffered glutaraldehyde separately and processed for light and electron microscopic studies, respectively. The reproductive system consisted of testis, commissure, vas deferens, ejaculatory duct and penis. The vas deferens was divided based on the morphology and/or histology into AVD, MVD and PVD. The AVD was further divided based on histology into proximal and distal regions, and the MVD, based on diameter into major and minor coils. The testicular lobe had several lobules with a central seminiferous tubule, which continued till the penis. The seminiferous tubule was lined by a layer of cuboidal or columnar epithelium. The lining of the central tubule of the vas deferens formed several ‘folds’, which at times formed ‘pouches’. High incidence of cell organelles in the columnar epithelial cells, aggregations of vesicles and occurrence of blebs at the luminal periphery and the projection of numerous microvilli containing electron‐dense materials into the lumen from the cell lining denoted high secretory activity of the epithelial cells.     

Variation in male reproductive traits among three bitterling fishes (Acheilognathinae: Cyprinidae) in relation to the mating system

Male traits that correlate with fertilization success include testis size and structure, ejaculate size, ejaculation frequency, and sperm motility. Two hypotheses potentially explain interspecific differences in these traits: sperm competition and sperm limitation. We examined variation in six traits associated with fertilization success in three closely‐related species of bitterling fish; the European bitterling (Rhodeus amarus), the Chinese rose bitterling (Rhodeus ocellatus), and the Chinese bitterling (Rhodeus sinensis). Interspecific differences indicated that the three study species have evolved different sperm allocation strategies. Rhodeus amarus displayed the most developed reproductive apparatus with a number of traits associated with both high levels of sperm production and fertilization efficiency. Rhodeus ocellatus and R. sinensis appear to have more comparable sperm allocation strategies, although relative testis size and spermatozoa head : tail ratio were greater in R. sinensis, suggesting that sperm competition risk may be higher in this species. All three species possessed an unusually well developed sperm duct with evidence of mucin production, which greatly extends the longevity of sperm and, consequently, the period over which fertilization can occur. We discuss these findings in the context of differences in the mating systems of the species examined, and relate the results obtained to differences in the temporal and spatial clustering of fertilizations. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 103 , 622–632.