Assessing the ovarian development of Macrobrachium amazonicum (Heller, 1862) phenotypes by means of an integrative analysis.

Macrobrachium amazonicum is a species of economic interest with a wide distribution in the Americas and high morphological and reproductive variability. Three phenotypes can be observed in this species: i) large-size amphidromous, ii) large-size and iii) small-size hololimnetic prawns. In the present work, the morphological, histochemical and ultrastructural aspects of ovarian development in the three phenotypes were comparatively analyzed. In addition, the interaction between the ovary and the hepatopancreas was investigated in these phenotypes through the use of gonadosomatic (GSI) and hepatosomatic (HSI) indices. Despite the morphological differences and different reproductive strategies adopted by the females, the macroscopic, histochemical and ultrastructural patterns of ovarian development showed no differences between the phenotypes. The ovaries were macroscopically classified into five stages of development (I to V). In early stages (I and II), the ovaries are full of oogonia, previtellogenic oocytes and oocytes in primary or endogenous vitellogenesis. At these stages, the rough endoplasmic reticulum (RER) produces a granular electron-dense material and sends it to the Golgi apparatus, where it will be modified, compacted and transformed into immature yolk granules. From stage III, secondary or exogenous vitellogenesis begins (with no interruption of endogenous vitellogenesis), where follicular cells nourish the oocytes and extracellular material is absorbed by endocytic vesicles, which fuse with immature yolk granules (forming mature granules) or with existing mature yolk granules. In stages IV and V, secondary vitellogenesis continues and mature yolk granules progressively occupy the cytoplasm. In M. amazonicum, the patterns of increase in oocyte diameter are quite similar between phenotypes, being greater in the small-size phenotype. This is related to the formation of larger oocytes/eggs and the production of large lipid reserves for their larvae. Changes in GSI and HSI during ovarian development show strong similarity between phenotypes, supporting the results obtained by histology and ultrastructure. Females in stages III and IV mobilize hepatopancreas reserves for ovarian maturation, which justifies the higher HSI values recorded in these stages. On the other hand, females in stage V show higher GSI and lower HSI values, indicating a mobilization of resources for the end of ovarian development as the females are ready to spawn.

The ultrastructure of spermatozoa of two species of Aegla (A. parana and A. quilombola) (Crustacea, Decapoda) endemic to Brazil.

The previously published ultrastructure of Aegla spermatozoa contributed to the phylogenetics of this unique taxon. The present study describes the spermatozoa of two additional aeglids, Aegla parana and A. quilombola. The spermatozoa consist of two hemispheres of the approximate same size and a bilayered acrosomal vesicle; both characteristics of the genus Aegla. The similarity of spermatozoa ultrastructure observed between A. parana and A. quilombola and the endemic Australian anomuran, Lomis hirta (Lomidae) reflects a sister group relationship, even though both are from different regions of the world and different environments today. Aeglid spermatozoa share the same organization with Lomis including the two equal size hemispheres separated by a membrane also two layers in the acrosomal vesicle with the external layer being surrounded by another membrane. The number of spermatozoa microtubular arms is unclear in Aegla, however, they are present in both the nucleus and cytoplasm. This observation does not agree with the presence of spermatozoa arms only in the nucleus, as an exclusive character for Aegla, as proposed previously. The presence of lipid-droplets and peroxisomes was observed only in the spermatozoa of A. quilombola. The greatly reduced number of spermatozoa observed in all specimens analyzed raises concerns about the conservation of several threatened species. In addition, the absence of any spermatophores seems to be a characteristic of the Aeglidae to date.

Male reproductive system of the freshwater prawn Macrobrachium brasiliense (Decapoda: Palaemonidae): Notes on spermatophore formation and sperm count.

Studies on the morphology of the reproductive system are essential for understanding the reproduction processes of species or even within genera or families. The present study aimed to describe the functional morphology of the male reproductive system, spermatophore formation, and sperm count of Macrobrachium brasiliense. The anatomy of the reproductive system consists of a pair of testes from which the vasa deferentia (VD) starts, extending to the fifth pair of pereopods. The VD is divided into three regions: proximal (PVD), middle (MVD), and distal (DVD). In the PVD, there is a prominent fold, the typhlosole, formed by columnar cells. The typhlosole disappears in the MVD, being incorporated into one of the faces of the VD wall, identified by its simple columnar epithelium while the remainder of the vessel wall is formed by squamous or simple cubic epithelium. Columnar cells produce type-II and III secretion. The epithelium in the DVD is made up only of cubic cells. Low sperm concentration was observed when compared to other species of the genus Macrobrachium. In conclusion, the typhlosole and columnar epithelium are responsible for the asymmetric spermatophore, which seems the pattern of Macrobrachium that is probably shared with other caridean shrimps.

The first zoeal stage of the deep-sea crab Tetraxanthus rathbunae (Brachyura: Pseudorhombilidae): a morphological description.

The description of the first zoea stage of Tetraxanthus rathbunae Chace, 1939, the third known zoea of Pseudorhombilidae and the first for Tetraxanthus, was carried out from laboratory hatched larvae. Their morphology differs from other pseudorhombilid species previously described primarily in the number and type of setae on the antennule, antennal exopod, maxilla, first maxilliped, and second maxilliped. These zoeae, however, share the same number of spines on the carapace (rostral, dorsal, and lateral), absence of the endopod on the antenna, long and well-developed spinose protopod, maxillary exopod with four marginal plumose setae and a robust posterior process, the absence of setae on the basis of the second maxilliped, and the presence of three lateral spines on each side of the furca. The results obtained in the present study are of great importance in being only the third known first stage pseudorhombilid zoea and the first for the Tetraxanthus, as well as in contributing to the identification of brachyuran planktonic zoea. Additionally, it also provides information that can help elucidate pseudorhombilid phylogeny.

Is there a trade-off between sperm production and sexual weaponry in the Amazon River prawn Macrobrachium amazonicum (Heller, 1862)?

The Amazon River prawn Macrobrachium amazonicum shows populations with four well-defined morphotypes in males. Dominant males of morphotypes green claw 1 (GC1) and green claw 2 (GC2) have large bodies and chelipeds and a higher reproductive success in comparison with the submissive morphotypes - translucent claw (TC) and cinnamon claw (CC). However, recently, some populations of the species do not have dominant morphotypes. Here, we compared the patterns of spermatic production and concentration among morphotypes and populations with three different phenotypes: (i) large-size amphidromous prawns, and (ii) large-size ("i" and "ii" with dominant morphotypes) and (iii) small-size hololimnetic prawns (without morphotypes). We described the spermatogenesis and the histochemical features of vasa deferentia (VD) and evaluated the relationship between the investment in spermatic production and sexual weapons acquisition in males of different phenotypes. The spermatic production and concentration in populations with four morphotypes were similar between morphotypes. The exception was the CC morphotype in which males had the seminiferous tubules filled with spermatocytes and low spermatic concentration. The spermatogenesis, spermiogenesis, and VD structure were not different among the studied phenotypes and populations. The seminal fluid of M. amazonicum is comprised by glycoproteins and by concentric layers of secretions of types I, II (basophilic), and III (eosinophilic). We could infer that males of dominant morphotypes allocate a higher amount of energy to the development of strong sexual weapons at the expense of the energy allocated to the reproductive system during the sequential growth. Inversely, small-size males direct more energy toward the spermatic production and transference at the expense of sexual weapons. Therefore, there is a clear trade-off between the investment in the gonadal and sexual weapons development in males of M. amazonicum.

Checklist of decapod crustaceans from the coast of the So Paulo state (Brazil) supported by integrative molecular and morphological data: V. Dendrobranchiata and Pleocyemata [Achelata, Astacidea, Axiidea, Caridea (Alpheoidea and Processoidea excluded), Gebiidea, Stenopodidea].

This checklist is the fifth and last compilation on the decapod crustaceans reported to So Paulo (Brazil) coastal area, resulting from long-term multidisciplinary projects, which combined morphological analyses and molecular techniques. The current research includes 75 decapod species, herein referred as shrimps/lobsters-like (shrimps, ghost-shrimps, lobsters, and related groups), reported to So Paulo coastal area. These species occur in marine, estuarine, and amphidromous habitats and are classified into 21 families as follow: Aristeidae (2 spp., 2 genera), Atyidae (4 spp., 2 genera), Axianassidae (1 sp., 1 genus), Callianassidae (1 sp., 1 genus), Callichiridae (6 spp., 4 genera), Crangonidae (1 sp., 1 genus), Glyphocrangonidae (1 sp., 1 genus), Luciferidae (2 spp., 2 genera), Nephropidae (4 spp., 2 genera), Palaemonidae (15 spp., 9 genera), Palinuridae (2 spp., 1 genus), Pandalidae (1 sp., 1 genus), Pasiphaeidae (1 sp., 1 genus), Penaeidae (10 spp., 6 genera), Sergestidae (3 spp., 3 genera), Sicyoniidae (4 spp., 1 genus), Scyllaridae (5 spp., 3 genera), Solenoceridae (4 spp., 3 genera), Stenopodidae (2 spp., 1 genus), and Upogebiidae (6 spp., 1 genus). We generated new sequences of cytochrome oxidase subunit I (barcode region) and 16S genes (51 and 54, respectively) of 54 species. Our examination concluded that 75 shrimps/lobsters-like species are reported to the So Paulo coast. We excluded Leander tenuicornis (Palaemonidae), Penaeus setiferus (Penaeidae), Philocheras gorei (Crangonidae), and Rhynchocinetes typus (Rhynchocinetidae) from this list.

From copula to fertilization: The reproduction of primitive crab Hypoconcha parasitica (Linnaeus, 1763) (Podotremata: Dromiidae).

This study describes the mating behavior of Hypoconcha parasitica under laboratory conditions highlighting the spermathecal morphology and focusing on the seminal fluid storage and release of spermatozoa. The pairs were kept in aquaria where the mating behavior was recorded and described. The spermathecae of the female were analyzed using scanning electron microscopy, X-ray micro-CT, histology, and histochemistry. No pre- or post-copulatory mate guarding was observed in H. parasitica. The sperm transfer occurred with each pair maintaining the protection shield (bivalve shell) on their dorsum. The pair of spermathecae is covered exclusively by the cuticle, following the Podotremata pattern. Many muscle fiber bundles are attached to the cuticular wall facing toward the cephalothorax cavity, especially covering the lateral and slightly dorso-anterior region toward the apodeme of Sternite 7. The spermathecal organization indicates that the process of sperm release during fertilization occurs through muscular action exerted by the female on the wall of the chamber. Thus, the musculature distribution in Hypoconchinae distinguishes them from described for other Podotremata such as Homolidae, which shows the musculature associated with the spermathecae aperture. Like the Homolidae, the first pleopod in H. parasitica seems to take part in the transfer/ movement of spermatozoa and oocytes between the sternum and abdomen, which forms a temporary chamber where fertilization occurs. In conclusion, the spermathecal morphology and associated structures bring new insights to the mechanisms involved in the sperm storage and fertilization of primitive crabs and how the Dromiidae spermatheca perform a new pattern among the poorly studied Podotremata.

Seminal fluid and spermatophore production in a western Atlantic invasive swimming crab, Charybdis hellerii, reveals a different pattern to Portunoidea.

The male reproductive system in Portunoidea is voluminous in order to produce the sperm plug, avoiding sperm competition. The portunid crab, Charybdis hellerii, is a successful invasive species, and this study describes the male reproductive system under light and electron microscopy and evaluates the gonadosomatic index and spermatophore dehiscence compared to other species that produce a sperm plug. The reproductive system is small, and its gonadosomatic index much lower than those of other Portunoidea that produce a sperm plug. The spermatophores are stored in the anterior part of the vas deferens, which is uncommon in Brachyura. Lateral outpocketings are found in the short, middle, and posterior parts of the vasa deferentia, where spermatophores are not found. The seminal fluid composition is different from the species that produce sperm plug. The usual portunid spermatophore wall is absent and the round coenospermic to even cleistospermic spermatophores are enclosed by secretions. This mucous-type spermatophore undergoes dehiscence in seawater or when passing through the gonopod. C. hellerii differ from other Portunoidea described regarding a low amount of seminal fluid production, spermatophore storage, and not show traits of producing sperm plugs. These different male reproductive features can lead to different female sperm storage in Portunoidea.

Much more than hooked: Setal adaptations for camouflage in Macrocoeloma trispinosum (Latreille, 1825) (Crustacea: Decapoda: Brachyura).

Several majoid crabs are known to adhere exogenous materials to their bodies, a behaviour called decoration. Until now, the adhesion of exogenous materials to the body is most attributed to the well-known hooked setae. Here, we analysed the carapace of Macrocoeloma trispinosum (Latreille, 1825) under light and electron microscopy to study the different mechanisms allowing majoid crabs to decorate themselves. Five setal types are described here, of which four for the first time: velvet type I, velvet type II, depressa and cattail seta. These setae are morphologically and histologically detailed, and new hypotheses about the fixation of exogenous material on the carapace are explored. M. trispinosum has a complex setal apparatus for the adhesion of the decoration, with tegumental ducts along the shaft of most setae. These tegumental ducts are connected to glands formed by large cells arranged radially (rosette or acini) at the base of the setae, in the connective tissue, just below the epithelium. We could observe these glands in different stages of maturation, and no valve-like structure was observed, which may indicate a continuous flow of protein secretion that could serve as an adhesive substance found in the apex of most setae. This is the first record indicating a potential chemical adhesion mechanism aiding the masking process in decorator crabs.

Long-Term Evaluation of Poly(lactic acid) (PLA) Implants in a Horse: An Experimental Pilot Study.

In horses, there is an increasing interest in developing long-lasting drug formulations, with biopolymers as viable carrier alternatives in addition to their use as scaffolds, suture threads, screws, pins, and plates for orthopedic surgeries. This communication focuses on the prolonged biocompatibility and biodegradation of PLA, prepared by hot pressing at 180 °C. Six samples were implanted subcutaneously on the lateral surface of the neck of one horse. The polymers remained implanted for 24 to 57 weeks. Physical examination, plasma fibrinogen, and the mechanical nociceptive threshold (MNT) were performed. After 24, 28, 34, 38, and 57 weeks, the materials were removed for histochemical analysis using hematoxylin-eosin and scanning electron microscopy (SEM). There were no essential clinical changes. MNT decreased after the implantation procedure, returning to normal after 48 h. A foreign body response was observed by histopathologic evaluation up to 38 weeks. At 57 weeks, no polymer or fibrotic capsules were identified. SEM showed surface roughness suggesting a biodegradation process, with an increase in the median pore diameter. As in the histopathological evaluation, it was not possible to detect the polymer 57 weeks after implantation. PLA showed biocompatible degradation and these findings may contribute to future research in the biomedical area.

Checklist of decapod crustaceans from the coast of the São Paulo state (Brazil) supported by integrative molecular and morphological data: IV. Infraorder Anomura: Superfamilies Chirostyloidea, Galatheoidea, Hippoidea and Paguroidea.

This checklist is the fourth contribution resulting from a long-term multidisciplinary project which combined morphological analyses and molecular techniques (mitochondrial DNA markers) for accurate identification of marine and coastal decapod crustaceans of São Paulo State (Brazil). We provide a list of 63 species of the following 11 families of 4 superfamilies of Anomura: Albuneidae (4 spp.), Blepharipodidae (1 sp.), Chirostylidae (1 sp.), Diogenidae (18 spp.), Hippidae (1 sp.), Munididae (8 spp.), Munidopsidae (1 sp.), Paguridae (13 spp.), Parapaguridae (2 spp.), Porcellanidae (13 spp.), and Pylochelidae (1 sp.). Seven species previously reported from the region were neither collected nor found in museum collections during our study, including one (Sympagurus dimorphus) that we suggest to be removed from São Paulo coast fauna lists. We generated new sequences of cytochrome oxidase subunit I (barcode region) and 16S genes of 44 species. This anomuran inventory may serve as guideline for future studies on taxonomy, conservation, population genetics, biogeography, and phylogenetics, which might flag species that deserve further investigations and concerns.

Is Potimirim potimirim (Crustacea, Decapoda, Atyidae) a protandric hermaphrodite species? Behavioral and morphological aspects of the reproductive system.

We evaluated the mating behavior of Potimirim potimirim in the laboratory based on the assumption that this caridean might be hermaphroditic. We also performed histology and scanning electron microscopy analyses of the reproductive system of females, males, and differentiated individuals. The mating experiments produced three behavioral stages, namely, interaction, lateral positioning, and copulation (which did not necessarily occur in this sequence). The hypothesis of a random pure searching mating was corroborated by the lack of male courtship, postcopulatory guarding, and the high aggregation of individuals. Three macroscopic ovarian stages were recorded in adult females: rudimentary, developing and mature. Secondary vitellogenesis begins at the developing stage, producing mature yolk. The male reproductive system is formed by testes and the vasa deferentia (VD), that is divided into three regions: proximal, middle, and distal. The proximal VD has a typhlosole that produces a thin layer of type II secretion around the central spermatozoa mass immersed in a type I secretion. External and adherent type III secretion is produced from the MVD to DVD, and both compound the primordial spermatophore. Potimirim potimirim has a gonochoric reproductive system and mating behavior and its sexual system does not fit into any of the previously described protandric systems.

Checklist of decapod crustaceans from the coast of the São Paulo state (Brazil) supported by integrative molecular and morphological data: III. Infraorder Brachyura Latreille, 1802.

This checklist is the third part of a series derived from a long-term multidisciplinary project on the biodiversity of decapod crustaceans from marine and coastal environments of São Paulo state (Brazil). We integrated molecular techniques (DNA markers) and morphological analyses of adult specimens for accurate identifications. We compilated 185 species from the literature, but we confirmed the presence of 168 species: 130 of which we sampled, analyzed and obtained sequences (COI and/or 16S totalizing 113 sequences) and 38 that were not directly collected but were confirmed by analyses. In addition, 27 had their distribution removed from São Paulo due to uncertainties, and absence of material as voucher. Five species were reported for the first time on the coast of São Paulo (Calappa ocellata, Neohelice granulata, Teleophrys pococki, Teramnonotus monodi, Tetraxanthus rathbunae) and one on the Brazilian coast (Pseudomedaeus agassizi). Most of the non-sampled species previously reported on the coast of São Paulo might be considered doubtful records stablished in the past by inaccurate analyses, which assumed extended distribution to the area and/or misidentifications. At this time and based on our refined compilation, we can estimate the brachyuran diversity on the coast of São Paulo in 168 species. This detailed inventory contributes to the knowledge on the local decapod fauna by checking available dataset, adding new species records in São Paulo and new sequences to GenBank database. These data may serve as baseline for future identifications and studies on conservation, population genetics, biogeography and phylogenetics, which might flag species that deserve further investigations and concerns.

Validation of Xiphopenaeus dincao Carvalho-Batista, Terossi, Zara, Mantelatto amp; Costa and Xiphopenaeus baueri Carvalho-Batista, Terossi, Zara, Mantelatto amp; Costa (Decapoda: Penaeidae) from western Atlantic.

Two species of Penaeidae shrimp from western Atlantic were described by Carvalho-Batista et al. (2019) as Xiphopenaeus dincao and Xiphopenaeus baueri. The descriptions and figures presented by Carvalho-Batista et al. (2019) and the posterior author's correction (Carvalho-Batista et al., 2020) characterize the species. However, the journal issue in which the description was published is online only, and the article with the new names did not include a Zoobank registration number (LSID), required for validation of new names in electronic-only publications (ICZN 2012). Thus, the names Xiphopenaeus dincao and Xiphopenaeus baueri, as published in Scientific Reports 9: 15281, is not available according the International Code of Zoological Nomenclature (ICZN 1999, 2012). Therefore, the present note serves to validate the names Xiphopenaeus dincao and Xiphopenaeus baueri by fulfilling ICZN conditions for nomenclatural availability. Type specimens of both species are deposited in Museu de Zoologia da Universidade de São Paulo (MZUSP), São Paulo, Brazil.

Ultrastructure and histochemistry of the male reproductive system of the genus Callinectes Stimpson, 1860 (Brachyura: Portunidae).

We described the ultrastructure and histochemistry of the reproductive system of five Callinectes species, and evaluate the seasonal variation in weight of the reproductive system and hepatopancreas by comparing annual changes of somatic indices. The somatic indices changed little throughout the year. In Callinectes, spermatogenesis occurs inside the lobular testes and, within each lobule, the cells are at the same developmental stage. Spermatogenesis and spermiogenesis follow the same development pattern in all Callinectes studied. Mature spermatozoa are released into the seminiferous ducts through the collecting ducts. Cells of the vas deferens are secretory as evidenced by rough endoplasmic reticulum, Golgi complex, and secretory vesicles that produce the seminal fluid. The anterior vas deferens shows two portions: proximal and distal. In proximal portion (AVDp), spermatozoa are clustered and embedded in an electron-dense, basophilic glycoproteinaceous secretion Type I. In the distal portion (AVDd), the spermatophore wall is formed by incorporation of a less electron-dense glycoproteinaceous secretion Type II. The secretion Type I change to an acid polysaccharide-rich matrix that separates the spermatophores from each other. The median vas deferens (MVD) stores the spermatophores and produces the granular glycoproteinaceous seminal fluid. The posterior vas deferens (PVD) has few spermatophores. Its epithelium has many mitochondria and the PVD seminal fluid changes into a liquid and homogeneous glycoprotein. Many outpocketings in the PVD and MVD help to increase the fluid production. Overall, the reproductive pattern of Callinectes is similar to other species that produce sperm plugs. The secretions of AVD, MVD, and PVD are responsible for the polymerization that forms the solid, waxy plug in the seminal receptacle. The traits identified here are common to all Portunidae species studied so far.

Size at onset of sexual maturity in Macrobrachium amazonicum (Heller, 1862) phenotypes: an integrative approach.

The size at the onset of sexual maturity (SOM) was determined in four hololimnetic populations of Macrobrachium amazonicum using morphometric, physiological and functional criteria. Male prawns from two populations analyzed had hypertrophied chelipeds and large body proportions, showing the presence of four morphotypes, i.e. large-size phenotype (LS). However, the other two populations showed smaller males without morphotypes, i.e. small-size phenotype (SS). The development of sexual weapons in males modulated the mating system and SOM in this species. It was verified that there is a synchrony between physiological and functional maturities in SS males. On the other hand, functional maturity in LS males occurred after physiological and morphological maturities. In females, we verified synchronization between morphological and functional maturities. The results obtained indicated that, in both sexes, physiological maturity anticipates the others criteria. There were no differences between the sequential scheme of sexual maturity for female phenotypes, being: physiological → functional morphological. However, the evaluation of sexual maturity in males must take into account the phenotype of population, since that was modulated by functional maturity. The sequential scheme for LS males is: physiological → morphological → functional, while for SS males it is: physiological = functional → morphological.

Author Correction: A multigene and morphological analysis expands the diversity of the seabod shrimp Xiphopenaeus Smith, 1869 (Decapoda: Penaeidae), with descriptions of two new species.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

A multigene and morphological analysis expands the diversity of the seabod shrimp Xiphopenaeus Smith, 1869 (Decapoda: Penaeidae), with descriptions of two new species.

After being stable for nearly a century, the taxonomic history of the genus Xiphopenaeus has been marked by many changes in the last three decades. The taxonomic status of the Atlantic species has a low resolution, and many species are still undefined and grouped as cryptic species. Here we employed an integrative approach to define the species of Xiphopenaeus and the morphological characters needed to differentiate them. We combined the analyses of two molecular markers (COI and 16 S rDNA), scanning electron microscopy and light microscopy. Based on specimens from 17 localities from the Atlantic and Pacific oceans, we detected five divergent genetic groups, three in the Atlantic (A1, A2, A3) and two in the Pacific (P1, P2). Male secondary sexual characters were able to differentiate four out of the five genetic groups. Group A1 corresponds to X. kroyeri, and A2 and A3 correspond to new species. We redescribed the genus and two new species are described and illustrated: Xiphopenaeus dincao nov. sp. (A2) and Xiphopenaeus baueri nov. sp. (A3). Since the holotype of X. riveti was missing and the specimen analysed from group P2 was a female, the status of the species of Xiphopenaeus from the Pacific remains unresolved.

Integrative depiction of the male reproductive system of the commercial purple crab Homalaspis plana (Platyxanthidae): Structure and function.

The information available on structures and function of reproductive systems are female-biased in many animal groups and crabs are not an exception. However, knowledge about the male reproductive system can help us to understand evolutionary and ecological processes such as the mating system and mating behavior. This study focuses on an integrative description of the principal regions of the male reproductive system of the purple crab, Homalaspis plana, a commercially exploited crab from the southeastern Pacific. For this description a combination of morphological observation, gravimetric records, histological and histochemical techniques and biochemical analysis were used. Also, a gross description of the seminal receptacle of the female was performed. The male reproductive system of H. plana was highly extended and complex, with a great capacity to produce and store spermatophores and secrete, at least, two kinds of seminal fluids. Sperm production, sperm packaging in spermatophores, storage of spermatophores and seminal liquid secretion were functions localized in specific regions. The vasosomatic index was the highest recorded to date in crabs, symptomatic of a high investment in reproduction. All these traits allow identification of H. plana as an internal sperm plug producer, which matches well with the highly extensible seminal receptacle of females. The great quantity of seminal material transferred to females during mating indicates that males may be able to mate only one time in the reproductive season and thus this species could be vulnerable to sperm depletion in males and sperm limitation in females if the fishery substantively reduced the abundance of males in the populations.

Parabronchial remodeling in chicks in response to embryonic hypoxia.

The embryonic development of parabronchi occurs mainly during the second half of incubation in precocious birds, which makes this phase sensitive to possible morphological modifications induced by O2 supply limitation. Thus, we hypothesized that hypoxia during the embryonic phase of parabronchial development induces morphological changes that remain after hatching. To test this hypothesis, chicken embryos were incubated entirely (21 days) under normoxia or partially under hypoxia (15% O2 during days 12 to 18). Lung structures, including air capillaries, blood capillaries, infundibula, atria, parabronchial lumen, bronchi, blood vessels larger than capillaries and interparabronchial tissue, in 1- and 10-day-old chicks were analyzed using light microscopy-assisted stereology. Tissue barrier and surface area of air capillaries were measured using electron microscopy-assisted stereology, allowing for calculation of the anatomical diffusion factor. Hypoxia increased the relative volumes of air and blood capillaries, structures directly involved in gas exchange, but decreased the relative volumes of atria in both groups of chicks, and the parabronchial lumen in older chicks. Accordingly, the surface area of the air capillaries and the anatomical diffusion factor were increased under hypoxic incubation. Treatment did not alter total lung volume, relative volumes of infundibula, bronchi, blood vessels larger than capillaries, interparabronchial tissue or the tissue barrier of any group. We conclude that hypoxia during the embryonic phase of parabronchial development leads to a morphological remodeling, characterized by increased volume density and respiratory surface area of structures involved in gas exchange at the expense of structures responsible for air conduction in chicks up to 10 days old.