PERFORATING POTENTIAL OF LOLIGINID SPERMATOPHORES

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ultrastructure of the spermatophores and spermatozoa from the gecarcinid land crab, Cardisoma guanhumi Latreille, 1828, (Crustacea, Brachyura, Gecarcinidae) from the western Atlantic

The ultrastructure of the sperm of the gecarcinid land crab Cardisoma guanhumi is described by scanning and transmission electron microscopy. Ultrastructural sperm morphology lends support for the monophyletic origin of the Thoracotremata, and for the placement of C. guanhumi in that clade. Additionally, it further attests the low level of variability within the thoracotreme clade. With regard to ultrastructural morphology, small differences concerning the apical button, the hyaline periopercular rim, the inner acrosome zone, and the outer two lamellations in the outer acrosome zone have been found between the spermatozoa of C. guanhumi and Cardisoma carnifex, the only other species in this genus studied for spermatozoal morphology. Copyright © 2013 Taylor & Francis.

Unraveling the structure of squids' spermatophores: a combined approach based on Doryteuthis plei (Blainville, 1823) (Cephalopoda: Loliginidae)

Marian, J.E.A.R. and Domaneschi, O. 2012. Unraveling the structure of squids spermatophores: a combined approach based on Doryteuthis plei (Blainville, 1823) (Cephalopoda: Loliginidae). Acta Zoologica (Stockholm) 93: 281307. Male coleoid cephalopods produce elaborate spermatophores, which function autonomously outside the male body during copulation, undergoing a complicated process of evagination. In order to contribute to the understanding of this unique structure, this study investigated the morphology of the spermatophore of Doryteuthis plei applying several microscopy techniques. A hitherto unreported, much more complex structural arrangement was revealed for the loliginid spermatophore, the most striking findings being: (1) the complex, layered structure of the middle membrane, which bears an additional, chemically distinct segment surrounding part of the cement body; (2) the presence of a space between the inner tunic and middle membrane filled with a fine reticulated material; (3) the presence of stellate particles not only embedded in the spiral filament, but also closely applied to the inner membrane at the level of the cement body; (4) the presence of a pre-oral chamber in the cap region; and (5) the complex organization of the cement body, formed by two distinct layers encompassing contents of different chemical and textural properties. Careful literature reassessment suggests several of these features are common to loliginids, and to some extent to other squids. Their possible functional implications are discussed in light of our knowledge of the spermatophoric reaction mechanics.

Revisiting a medical case of "stinging" in the human oral cavity caused by ingestion of raw squid (Cephalopoda: Teuthida): new data on the functioning of squid's spermatophores

Male squid produce intricate spermatophores that, when transferred to the female, undergo the spermatophoric reaction, a complex process of evagination that leads to the attachment of the spermatangium, that is, the everted spermatophore containing the sperm mass. While this process is still not completely understood, the medical literature includes several reports of "oral stinging" (i.e., punctured wounds in the human oral cavity) following consumption of raw male squid, which contains undischarged spermatophores able to inflict such wounds. Here, we revisit a recent medical report of oral stinging by Shiraki et al. (Pathol Int 61:749-751, 2011), providing an in-depth reanalysis of their histological biopsies and revealing vital information on the functioning of squid spermatophores. The morphology of the spermatangia attached within the oral cavity is similar to the condition found in spermatangia naturally attached to female squids. The spermatangia were able to superficially puncture the superficial layers of the oral stratified squamous epithelium, and numerous, minute stellate particles from the squid spermatophore were found adhered to the oral epithelium. These findings corroborate previous hypotheses on the functioning of squid spermatophores, namely that spermatophore attachment generally involves tissue scarification, and that stellate particles play a vital role in the attachment process. Moreover, spermatophore attachment is confirmed to be autonomous (i.e., performed by the spermatophore itself) in another squid species (possibly a loliginid), and the results strongly indicate that the attachment mechanism is not dependent upon a specialized epithelium, nor a mate's specific chemical stimulus. From the pathological point of view, the best prophylactic measure at present is the removal of the internal organs of the raw squid prior to its consumption.

On the structure of the seminal receptacle in cyclopids (Copepoda, Cyclopoida). [Translation from: Informatsionnyi Byulleten Biologiya Vnutrennikh Vod (6) 26-31, 1970.]

The seminal bag, or seminal receptacle, forms a characteristic organ of cyclopids, serving for retention of the sperms discharged from the spermatophores. The structure of the seminal receptacle, more precisely its form, is fairly widely used in diagnosis and undoubtedly can be more widely applied in the systematics of the group. Within the limits of the family Cyclopidae it is possible to distinguish crustaceans with three basic types of seminal bag. The differences consist of the position which this organ occupies in the genital segment. of one species, we carried out a series of observations on its formation in ontogenesis and during the life of the adult stage. As material for observation the study used laboratory cultures of three species; Acanthocyclops americanus (Marsh) from the plankton of the Moscow River, Cyclops vicinus Uljan and Mesocyclops leuckarti Glaus from the plankton of the channel section of the upper part of the Gorkovsk reservoir. The author concluded that the irreversibility of the changes in the seminal receptacle presents the possibility of utilising this structure as one of the indicators of the growth of the individual.

Length at maturity in three pelagic sharks (Lamna nasus, Isurus oxyrinchus, and Prionace glauca) from New Zealand

Reproductive data collected from porbeagle, shortfin mako, and blue sharks caught around New Zealand were used to estimate the median length at maturity. Data on clasper development, presence or absence of spermatophores or spermatozeugmata, uterus width, and pregnancy were collected by observers aboard tuna longline vessels. Direct maturity estimates were made for smaller numbers of sharks sampled at recreational fishing competitions. Some data sets were sparse, particularly over the vital maturation length range, but the availability of multiple indicators of maturity made it possible to develop estimates for both sexes of all three species. Porbeagle shark males matured at 140–150 cm fork length and females at about 170–180 cm. New Zealand porbeagles therefore mature at shorter lengths than they do in the North Atlantic Ocean. Shortfin mako males matured at 180–185 cm and females at 275 –285 cm. Blue shark males matured at about 190 –195 cm and females at 170–190 cm; however these estimates were hampered by small sample sizes, difficulty obtaining representative samples from a population segregated by sex and maturity stage, and maturation that occurred over a wide length range. It is not yet clear whether regional differences in median maturity exist for shortfin mako and

Reproductive biology of the blue swimmer crab (Portunus pelagicus, Decapoda: Portunidae) in five bodies of water on the west coast of Australia

Portunus pelagicus was collected at regular intervals from two marine embayments and two estuaries on the lower west coast of Australia and from a large embayment located approximately 800 km farther north. The samples were used to obtain data on the reproductive biology of this species in three very different environments. Unlike females, the males show a loosening of the attachment of the abdominal flap to the cephalothorax at a prepubertal rather than a pubertal molt. Males become gonadally mature (spermatophores and seminal fluid present in the medial region of the vas deferentia) at a very similar carapace width (CW) to that at which they achieve morphometric maturity, as reflected by a change in the relative size of the largest cheliped. Logistic curves, derived from the prevalence of mature male P. pelagicus, generally had wider confidence limits with morphometric than with gonadal data. This presumably reflects the fact that the morphometric (allometric) method of classifying a male P. pelagicus as mature employs probabilities and is thus indirect, whereas gonadal structure allows a mature male to be readily identified. However, the very close correspondence between the CW50’s derived for P. pelagicus by the two methods implies that either method can be used for management purposes. Portunus pelagicus attained maturity at a significantly greater size in the large embayment than in the four more southern bodies of water, where water temperatures were lower and the densities of crabs and fishing pressure were greater. As a result of the emigration of mature female P. pelagicus from estuaries, the CW50’s derived by using the prevalence of mature females in estuaries represent overestimates for those populations as a whole. Estimates of the number of egg batches produced in a spawning season ranged from one in small crabs to three in large crabs. These data, together with the batch fecundities of different size crabs, indicate that the estimated number of eggs produced by P. pelagicus during the spawning season ranges from about 78,000 in small crabs (CW=80 mm) to about 1,000,000 in large crabs (CW=180 mm).

A study of the male reproductive organs of nine species of penaeid prawns (Crustacea: Penaeinae)

This paper deals with the male reproductive organs of nine species of penaeid prawns; Penaeus penicillatus Alcock, P. merguiensis De Man, P. semisulcatus De Haan, Metapenaeus affinis (H.Milne Edwards), M. monoceros (Fabricius), M. stebbingi Nobili, Parapenaeopsis hardwickii (Miers), P. sculptilis (Heller) and P. stylifera (H. Milne Edwards). The male reproductive organs exhibited structural variations, which were more pronounced at generic level. These variations are mainly due to the type of spermatophore they possess. One species of each genus, that is, P. merguiensis, M. affinis and P. sculptilis were also studied histologically to examine the internal structure of the male reproductive organs. Spermatophores of the six species belonging to the genera Penaeus and Parapenaeopsis are also described and illustrated.

Artificial insemination in penaeid shrimps

Artificial Insemination (AI) is a tool for genetic manipulation in the shrimp stocks. It is seen as one of the means for propagating shrimp culture to new areas by controlled reproduction. Attempts at artificial insemination in the dominant closed-thelycum penaeid shrimps species of the area viz. Metapenaeus affinis and Metapenaeus brevicornis were induced in wild adult stocks collected off Mumbai coast. Female specimens were subjected to unilateral eyestalk ablation by pinching so as to induce moulting and maturation. AI was performed two days after moulting on these females when the cuticle was still soft and flexible. Moulting also ensured rejection of initial spermatophores, if present. Response of males to electrical stimulation for spermatophore expulsion was spontaneous. Use of tissue glue for spermatophore retention was found to be unnecessary. Latency period ranged between10-16 days, while spawning occurred within 10-12 days of spermatophore transfer. Three partial spawning were recorded viz., two in Metapenaeus affinis and one in Metapenaeus brevicornis with an average spawning and hatching rates of 30% and 72.3% respectively. Average survival from first nauplius (N1) to one-day old post-larva (PLI) was a meager 3.43%. Use of AI in genetic manipulation of shrimp stocks for aquacultural purposes is indicated.

Gonad development characteristics and sex ratio in triploid chinese shrimp (Fenneropenaeus chinensis)

This paper details for the first time the gonad development characteristics and sex ratio of triploid shrimp (Fenneropenaeus chinensis). In triploid shrimp the development of gonad is apparently impaired, especially in females. In the ovary of triploids, germ cells mainly remain at oogonia stage during September through December. From January to February of the next year, partial primary oocytes developed in the ovary lobes. Spermatocytes and spermatids could be observed in the testes of triploids, and a few sperm were observed in the vas deferens and spermatophores. The morphology of sperms in triploid shrimp was abnormal. Flow cytometry was used to detect the ploidy of sperm in the vas deferens. The data showed that triploidy could affect the sex ratio in Chinese shrimp. The female-to-male ratio in triploids of about 4:1 will favor triploid shrimp aquaculture.

The adaptive significance and prevalence of courtship feeding in Hawaiian swordtail crickets

Males of many insect species feed their partner during courtship and mating. Studies of male nutrient donation in various systems have established that nuptial feeding has evolved mostly through sexual selection. Although there is extensive diversity in form, the function of nuptial gifts is typically limited to either facilitating copulation or increasing ejaculate transfer, depending on the time at which the gift is consumed by females. Unlike other insects, the Hawaiian swordtail cricket Laupala (Gryllidae: Trigonidiinae) exhibits serial transfer of nuptial gifts. Males transfer multiple spermless 'micro' spermatophores over several hours before mating at the end of the day (i.e. before the transfer of a single sperm-containing 'macro' spermatophore). By experimental manipulation of male microspermatophore donation, I tested several hypotheses pertaining to the adaptive significance of nuptial gifts in this system. I found that microspermatophore transfer improves insemination, by causing the female reproductive tract to take in more sperm. This result reveals a previously undocumented function for premating nuptial gift donation among insects. Enhanced sperm transfer due to microspermatophore donation may represent male manipulation or an internal mechanism of post-copulatory choice by females. I also performed experimental manipulation of male photoperiod to investigate how time and gender influence nuptial gift production and mating behavior. I found that the timing of mating is limited in males but not females and that the time of pair formation has consequences for the degree of nuptial gift donation, which suggests that both mating timing and microspermatophore number is important for male reproductive success. Finally, I observed the mating behavior of several trigonidiine taxa for a comparative analysis of sexual behavior and found that other genera also utilize spermless microspermatophores, which suggests that microspermatophore donation may be a common nuptial gift strategy among swordtail crickets. The elaborate nuptial feeding behavior of Hawaiian swordtail crickets prior to mating represents a newly discovered strategy to increase male insemination success rather than mating success. Based on this unexpected result, it is worth exploring whether courtship behaviors in other cricket or insect mating systems have also evolved to increase sperm uptake.

Some studies on the reproductive endocrinology of tiger prawn Penaeus monodon Fabricius

The growing demand for quality prawn seed from the farmers‘ and entrepreneurs, coupled with uncertainity of their availability from nature at the appropriate time in required quantities has prompted‘ research on problems connected with prawn seed production. Endocrine control of reproduction in the penaeid shrimp _P_. monodon has been investigated in detail by adopting a comprehensive approach to the problem. The major aspects of the study included in depth investigations of the cytological details of the reproductive and neuroendocrine organs in correlation with the process of gonadal maturation. Based on the conclusions drawn from such ultrastructural studies various endocrine manipulations were carried out to see their effects on gonadal maturation. Besides that studies on the reproductive quality of male P_. monodon and the cryopreservation of spermatophores form a part of the present investigation. The shrimp 3; Inonodon used in the present study were collected from the offshore waters of Cochin, Madras and Mandapam and from the culture ponds of Vypeen Island near Cochin (Kerala) . The entire investigation on the cytological aspects were carried out using standard histological and electron-microscopic methods. Endocrine manipulations and cryopreservation studies is also carried out using the standard methods.

Studies on the female reproductive physiology of the prawn, Macrobrachium idella (Hilgendorf, 1898)

’l‘he reproductive physiology of the female palaemonid prawn M. idella has been investigated by adopting a comprehensive approach to the problem. The major aspects of the study included investigations on breeding biology and process of oogenesis, variations in the biochemical components in relation to maturation, neuroendocrine relations and control over reproduction, and artificial insemination. The prawns used in the present study were procured from Vembanad Lake at Panavally village - a place nearly 20 km. away from Cochin. The studies were carried out using standard histological and biochemical methods. The modern technique of electroejaculation was adopted for extrusion of spermatophores in artificial insemination experiment.

Spermatogenesis in the blue swimming crab, Portunus pelagicus, and evidence for histones in mature sperm nuclei

Spermatogenesis in the blue swimming crab, Portunus pelagicus, is described by light and electron microscopy. The testis is composed of anterior (AT) and posterior (PT) lobes, that are partitioned into lobules by connective tissue trabecula, and further divided into zones (germinal, transformation and evacuation), each with various stages of cellular differentiation. The vas deferens is classified into three distinct regions: anterior (AVD), median (MVD), and posterior (PVD), on the presence of spermatophores and two secretions, termed substance I and II. Based on the degree and patterns of heterochromatin, spermatogenesis is classified into 13 stages: two spermatogonia (SgA and SgB), six primary spermatocytes (leptotene, zygotene, pachytene, diplotene, diakinesis, and metaphase), a secondary spermatocyte (SSc), three spermatids (St 1–3), and a mature spermatozoon. Spermatid stages are differentiated by chromatin decondensation and the formation of an acrosomal complex, which is unique to brachyurans. Mature spermatozoa are aflagellated, and have a nuclear projection and a spherical acrosome. AUT-PAGE and Western blots show that, during chromatin decondensation, there is a reduction of most histones, with only small amounts of H2B and H3 remaining in mature spermatozoa.

Spermathecae of the mangrove crab Ucides cordatus: a histological and histochemical view

Ucides cordatus is the most commercially, important mangrove crab in Brazil. In spite of its economic importance, there are few studies of its reproduction, in particular the female reproductive system. The present study describes the histology and histochemistry of the spermathecae of U. cordatus. Adult females were can lit monthly from July 2004 through June 2005, at Iguape, State of São Paulo. The crabs were anaesthetized, and their spermathecae removed and fixed in Davidson's fluid, following the histological routine for paraffin. The slides were stained with HE, xylidine Ponceau, PAS, alcian blue (pH 1.0 and 2.5), Sudan black B and picrosirius-haematoxylin. Histologically, the spermathecae possesses a capsule of conjunctive tissue, rich in collalgen fibres, which surrounds the secretory columnar epithelium. In the lumen, individual sperm packets are not observed; the spermatophores are intermixed with the seminal fluid and secretions of the spermathecae itself. A large proportion of the free spermatozoids and spermatophores are arranged in homogeneous masses in the proximal part of the spermathecae. The secretion produced by the columnar epithelium appears to promote the movement of the gametes to the fertilization chamber, in a ventral position, allowing fertilization of the oocytes. Histochemically, the secretion produced by the columnar epithelium was strongly positive for neutral polysaccharides, positive for acid polysaccharides, and weakly positive for proteins and lipids. Tills secretion forms a glycoprotein matrix which is associated with maintenance of the spermatophores, which can remain stored for long periods.