Elasmobranch Reproduction Research Articles

Long-term monitoring of egg-laying cycle using ultrasonography reveals the reproductive dynamics of circulating sex steroids in an oviparous catshark, Scyliorhinus torazame

The many diverse reproductive strategies of elasmobranchs (sharks, skates and rays) from lecithotrophic oviparity to matrotrophic viviparity have attracted significant research attention. However, the endocrine control of elasmobranch reproduction is less well-documented largely due to their reproductive characteristics, such as a long reproductive cycle, and/or repeated internal fertilization using stored sperm in oviparous species. In the present study, for the first time, we succeeded in non-invasive monitoring of the continuing egg-laying cycle of the cloudy catshark Scyliorhinus torazame using portable ultrasound devices. Furthermore, long-term simultaneous monitoring of the egg-laying cycle and measurement of plasma sex steroids revealed cycling patterns of estradiol-17β (E2), testosterone (T) and progesterone (P4). In particular, a decline in T followed by a reciprocal surge in plasma P4 were consistently observed prior to the appearance of the capsulated eggs, implying that P4 is likely associated with the ovulation and/or egg-case formation. While the cycling pattern of E2 was not as apparent as those of T and P4, threshold levels of E2 (>5 ng/mL) and T (>1 ng/mL) appeared to be crucial in the continuation of egg-laying cycle. The possibility to trace the dynamics of plasma sex steroids in a single individual throughout the reproductive cycles makes the catshark a useful model for regulatory and mechanistic studies of elasmobranch reproduction.

Re-evaluation of reproductive cycle and fecundity of finetooth sharks Carcharhinus isodon (Valenciennes 1839) from the Northwest Atlantic Ocean, with new observations on ovarian cycle and reproductive endocrinology of biennially reproducing sharks.

To provide updated information on life history for improved fishery management, the reproductive cycle of the finetooth shark Carcharhinus isodon from Northwest Atlantic (NWA) populations was investigated by examining temporal changes in morphology and histology of reproductive organs. Changes in plasma concentrations of gonadal sex hormones in relation to reproductive stage were also examined. Increases in testis width, epididymis head width, plasma testosterone concentrations and occurrence of mature spermatozoa were observed in male sharks between December and April, suggesting a seasonal pattern in reproduction that culminates with copulatory activity in May. Increases in maximum follicular diameter, oviducal gland width, plasma 17β-estradiol concentrations and occurrence of vitellogenic follicles were observed in non-pregnant female sharks during the same time period along with the occurrence of newly pregnant females in May, demonstrating strong synchronicity between male and female reproductive cycles. Pregnant females bearing full-term embryos were also observed in May, indicating that parturition occurs between mid-May and early June and gestation requires 12 months. Only transient temporal changes in follicle size and oviducal gland width were observed in pregnant females, indicating that reproductive periodicity is biennial; nonetheless, a single female exhibiting signs of concurrent vitellogenesis and pregnancy was observed. Mean brood size ± S.D. was 3.9 ± 0.9 offspring/female. Fecundity was not significantly correlated with female size, in part due to an unexpectedly high rate of early embryo mortality, which occurred in 11% of pregnant females, and was more common in larger individuals. Changes in ovarian activity during mid-pregnancy were observed, suggesting possible roles for the ovary in regulating some aspects of early to mid-gestation. This study confirms that earlier characterizations of the reproductive cycle and fecundity in NWA finetooth sharks remain valid for use in fishery management. This study also highlights unusual features of finetooth shark pregnancy (e.g., early embryo death, mid-pregnancy changes in ovarian function) that may have broader relevance to understanding elasmobranch reproduction.

Anthropogenic stressors influence reproduction and development in elasmobranch fishes

The consequences of human influence can arise in vertebrates as primary, secondary, or even tertiary stressors and may be especially detrimental for slow growing species with long generation times (i.e., K-selected species). Here, we review the impacts of both direct and indirect human interactions on the reproductive biology of elasmobranchs. Within direct human influence, capture-induced stress from fisheries bycatch and poor coastal management practices leading to habitat destruction and pollution are among the most impactful on elasmobranch reproduction. Capture-induced stress has been shown to negatively influence offspring and reproductive capacity via capture-induced parturition as well as by disrupting the reproductive physiology of adults. Habitat degradation impacts essential ecosystems that are necessary for the development of young elasmobranchs. Pollutants such as heavy metals, legacy pesticides, and flame retardants have been traced through elasmobranch reproduction; however, the long-term effects of these exogenous chemicals are yet to be determined. Furthermore, within indirect human impacts, climate change-mediated influences (e.g., ocean warming and acidification) can impact development, physiological processes, and behavioral patterns necessary for essential tasks such as foraging, growth, reproduction, and ultimately survival. Here, we also present a case study, where data regarding temperature and incubation time from 28 egg-laying elasmobranch species were examined to show relevance of such data in predicting how suitable (e.g., via maximum threshold temperatures) habitats will be for skate and shark development in the coming century. Concomitantly, this information highlights areas for future research that will help inform better management as well as climate change forecasting for this threatened group of aquatic vertebrates.

Reproductive Science in Sharks and Rays.

Sharks and rays make up 96% of the class Chondrichthyes. They are among the most endangered of any taxa, threatened through habitat loss, overfishing and hunting for shark fin soup, traditional medicines or sport, and because many species are slow to mature and produce low numbers of offspring. Sharks and rays are ecologically and reproductively diverse, though basic knowledge of their reproductive physiology is lacking for many species. There has been a move towards non-lethal approaches of data collection in sharks and rays, especially with reproductive technologies such as ultrasound and hormone analysis. Additionally, technologies such as semen collection and artificial insemination are lending themselves to develop tools to manage small or closed populations, with cold-stored sperm being shipped between institutions to maximize genetic diversity in managed populations. The role of steroid hormones in elasmobranch reproduction appears broadly conserved, though heavily influenced by environmental cues, especially temperature. For this reason elasmobranchs are likely at risk of reproductive perturbations due to environmental changes such as ocean warming. Current reproductive technologies including computer assisted sperm assessments to study warming effects on sperm motility and intra-uterine satellite tags to determine birthing grounds will serve to generate data to mitigate anthropogenic changes that threaten the future of this vulnerable groups of fish.

Evaluation of the use of portable ultrasonography to determine pregnancy status and fecundity in bonnethead shark Sphyrna tiburo.

This study examined the usefulness of portable ultrasonography in accurately predicting pregnancy and fecundity in the bonnethead shark Sphyrna tiburo by comparing ultrasound-obtained data with those obtained from animal dissection, the gold standard for elasmobranch reproduction studies. Mature female S. tiburo (n = 66) were collected throughout the period of reproduction and examined via portable ultrasonography using two different ultrasound transducers (8-5 MHz linear array transducer and a 5.0-2.5 MHz curvilinear array transducer) to determine pregnancy status and fecundity. Ultrasound-derived data were compared with validated assessments of pregnancy and litter size obtained using animal dissection to determine the level of agreement between the two approaches. Overall, there was strong agreement (90.9%) between pregnancy determinations obtained using ultrasonography and dissection. However, the effectiveness of portable ultrasonography in accurately determining specific stages of maturity and pregnancy differed slightly between transducer types (linear = 61.3%; curvilinear = 88.9%). Measurements of fecundity also agreed well between ultrasonography and dissection and there were no significant differences between mean fecundity estimates obtained using the two methods. In general, portable ultrasonography was found to be a good alternative to lethal sampling and animal dissection in obtaining reproductive data critical for the management and conservation of elasmobranchs. Special attention should be given to factors that can influence the reliability of ultrasound-derived data such as transducer type, embryo size and sound wave interference due to gut contents.

Impacts of fisheries on elasmobranch reproduction: high rates of abortion and subsequent maternal mortality in the shortnose guitarfish

AbstractThe fate of bycatch species released back into the water is crucial to estimate population status and consequent decisions to implement adequate management measurements for elasmobranchs. The shortnose guitarfish, Zapteryx brevirostris, is an endemic species of the Southwest Atlantic. It is currently listed as ‘Vulnerable’ both in the International Union for Conservation of Nature and Brazilian red lists. Although this species displays high tolerance to capture and transportation by fishers from the capture site to the fish market, a significant reduction in survival rates is observed during the reproductive period. Thus, the aim of the present study was to examine post‐capture and post‐commercial transportation consequences in pregnant females obtained as bycatch from artisanal fisheries in Southern Brazil. Results showed high abortion rates and reduced maternal survival after abortion, with all abortions occurring within 28 h of monitoring in laboratory. Stress plasma markers were also evaluated, to determine the physiological consequences of capture‐induced parturition to pregnant females. Results showed alteration in all markers measured (plasma urea, pH, lactate, phosphorus, and potassium) with values for dead pregnant females being different from non‐pregnant females and those which survived capture and subsequent abortions. The mortality caused by fisheries bycatch may prove difficult to manage, with immediate release and specific handling protocols for pregnant females implemented through environmental education environmental education and fishermen training programs, potential options to maintain adequate maternal survival and recruitment for this species.

Diagnostic assessment of reproductive status in white-spotted bamboo sharks (Chiloscyllium plagiosum)

Relatively little is known about elasmobranch reproductive physiology compared to other species. An increased understanding of elasmobranch reproduction would improve the success of captive breeding and may aid in situ conservation efforts by reducing demand for wild-caught individuals. The reproductive status of seven adult female white-spotted bamboo sharks (Chiloscyllium plagiosum) (WSB) was monitored via coelomic ultrasonography and analysis of plasma biochemistry and steroid hormones over 6 months. Based on ultrasonic findings, females were categorized at each blood collection time point as: no follicular activity (INACTIVE), follicles but no eggs present (ACTIVE-OVARY), and eggs present within the oviduct (ACTIVE-OVIDUCT). Triglyceride concentrations were greater for those with the ACTIVE-OVARY (75.98 mg/dL; CI 61.81–90.15 mg/dL) and ACTIVE-OVIDUCT (87.0 mg/dL; CI 70.20 to 103.81 mg/dL) categories than INACTIVE (51.81 mg/dL; CI 37.07–66.54 mg/dL) category. No significant differences were observed for PCV, total solids, calcium, phosphorus, iron or progesterone. Estradiol concentrations were less for the INACTIVE (0.15 ng/ml; CI 0.08 to 0.25 ng/ml) than ACTIVE-OVARY (0.63 ng/ml; CI 0.42 to 0.88 ng/ml) and ACTIVE-OVIDUCT (0.92 ng/ml; CI 0.64–1.26 ng/ml) category. Testosterone concentrations differed among reproductive states, being greater with the INACTIVE (0.22 ng/ml; CI 0.13 to 0.37 ng/ml) and peaking in the ACTIVE-OVIDUCT (2.12 ng/ml; CI 1.25–3.60 ng/ml) state. The ultrasonic technique was performed in a standardized manner, and the anatomy was validated via opportunistic post-mortem examinations and MRI. Using the described diagnostic techniques, reproductive status in WSB can be routinely monitored, and findings have implications for improving the success of captive breeding efforts in other elasmobranch species.

Changes in sex steroid hormone levels reflect the reproductive status of captive female zebra sharks (Stegostoma fasciatum)

Captive breeding in aquaria is a useful means for ex situ preservation of threatened elasmobranch species. To promote captive breeding, it is important to determine the female reproductive status. However, information regarding reproductive status in female elasmobranchs is limited. Here, we used zebra sharks, Stegostoma fasciatum, as a model for elasmobranch reproduction in captivity. We investigated the relationships among changes in the sex steroid hormone levels, follicle size, and egg-laying period to develop indicators for the female reproductive status. We confirmed that mature female zebra sharks undergo an annual reproductive cycle. Additionally, we showed that the variations in sex steroid hormone levels correlated with reproductive status in mature female zebra sharks. Plasma estradiol-17ß (E2) concentrations increased two months before ovarian follicle development and decreased along with follicle regression. Interestingly, E2 levels were inversely correlated with water temperature (R = −0.901). Moreover, high levels of testosterone (T) correlated well with the laying period. These results strongly suggest that E2 is an indicator for ovarian follicle development, and that T is a useful indicator for both the onset and end of the egg-laying period in captive zebra sharks.

Development of a Nonlethal and Minimally Invasive Protocol to Study Elasmobranch Reproduction

AbstractAn understanding of basic reproductive biology is essential for successful species‐specific management of elasmobranch fishes (sharks, skates, and rays). Such information is often gained through gross dissection or other lethal techniques, which are not appropriate for threatened and endangered species. Previous work on other vertebrates suggested that sex steroid hormones can be extracted from muscle tissues to identify reproductive status. Collecting for muscle biopsy is quick and minimally invasive and can be done without removing an animal from the water. Thus, the objective of the current study was to determine the efficacy of using muscle steroid hormones to assess the reproductive biology of elasmobranch fishes. The results suggest that concentrations of muscle progesterone, testosterone, and estradiol can be successfully quantified to study reproduction by radioimmunoassay. Additionally, there were significant correlations between the plasma and muscle estradiol concentrations in Spiny Dogfish Squalus acanthias and the progesterone, testosterone, and estradiol concentrations in Atlantic Sharpnose Sharks Rhizoprionodon terraenovae. The present investigation thus demonstrates that skeletal muscle is a nonlethally harvestable tissue that is well suited for studying the reproductive biology of elasmobranchs.Received October 12, 2012; accepted March 12, 2013

Calcitonin-like immunoreactivity in serum and tissues of the bonnethead shark, Sphyrna tiburo.

Calcitonin is a 32-amino acid peptide hormone that is best known for its actions in maintaining skeletal integrity and calcium homeostasis in mammals. Calcitonin also appears to function in regulating certain aspects of animal reproduction, but the nature of this role remains unclear, particularly in nonmammalian vertebrates. The present study investigated the relationship between calcitonin and reproduction in the bonnethead shark (Sphyrna tiburo), a well-studied member of the oldest living vertebrate group (i.e. elasmobranchs) known to possess a calcitonin-producing organ. Serum calcitonin concentrations were measured in 28 reproductively mature female S. tiburo using a heterologous enzyme-linked immunosorbent assay (ELISA) system. Sites of calcitonin immunoreactivity were detected in tissues of mature female and embryonic S. tiburo using immunocytochemistry. Significant increases in serum calcitonin concentrations of mature female S. tiburo occurred during early stages of gestation, a period characterized by yolk-dependency of developing embryos. Immunoreactive calcitonin was detected in the duodenum and pancreas of embryonic S. tiburo sampled during the same period. The results from this study suggest that calcitonin obtained from endogenous and/or maternal sources may function in regulating yolk digestion in embryonic S. tiburo. Therefore, the association between calcitonin and reproduction in elasmobranchs may reflect an important role for this hormone in embryonic development.

Putative Mating Behavior in Basking Sharks off the Nova Scotia Coast

We report aerial observation of 13 basking sharks (Cetorhinus maximus) engaged in putative mating behavior near the ocean surface 21 June 1998 at approximately 1230 AST, 140.8 km from the Nova Scotia mainland in the North Atlantic at 44?16'N, 61038'W, depth 130 m. Video and photo documentation showed the 68 m long sharks, some with discrete white patches on the dorsum and appendages, swimming slowly clockwise in a tight circle with many animals closely associated with up to three other animals. Previously described behaviors and motor patterns consistent with elasmobranch reproduction including parallel swimming, possible biting, nudging, and male on top of female were seen as well as other behaviors, including nose-to-tail following; flank approach; close approach involving rostral contact or proximity with the gill, pectoral fin, vent, and dorsum; and echelon swimming. Behaviors involving close and persistent physical association of two or more animals and the persistence of the circling behavior in a circumscribed area over a period in excess of 5 min indicated that this may represent the first well-documented observation of group courtship behavior in basking sharks.

Review. Levels of Recirculating Reproductively-related Steroid Hormones in Female Elasmobranchs. Implications for Reproduction in a Captive Environment

Chondrichthyans are a diverse group in terms of reproductive mode, which range from extended oviparity to placental viviparity. Although the reproductive biology of several species has been investigated, the reproductive physiology of only a few species has been reported. Only recently have the physiological factors of reproduction in cartilaginous fishes received attention. The information gained on reproductive cycles (both gonadal and uterine) has shown some similarities as well as differences between species. The serum levels of androgens, oestrogens, and progestins have been examined in several elasmobranch species and it has been shown that they are important in regulating key events in reproduction. The endocrine control of reproduction in elasmobranchs is more similar in many ways to higher vertebrates such as mammals than to teleosts. In general, the role of progesterone becomes more important than that of oestrogen in the transition from oviparity to viviparity. In addition, the roles of peptide hormones such as relaxin and the neurohypophysial hormones become increasingly important in viviparous species as in higher vertebrates. It is vital to understand the environmental and hormonal control of reproduction in elasmobranchs if more success is to be achieved in their reproduction in captivity. Knowledge gained from studies of the endocrine regulation of the reproductive cycle can be used to enhance the reproductive success of captive sharks, skates, and rays.

Endocrine regulation of the elasmobranch reproductive tract

AbstractAlthough the rich variety of reproductive tract adaptations in elasmobranchs has been well documented and involves many variants associated with oviparity and extensive evolution of viviparity in the group, little is known of the hormone dependence of reproductive tract development, differentiation, and function in the adult. In this paper, we present a brief review of some of our observations on the subject made over the past 15 years in the context of earlier observations. In total, despite the paucity of information, the observations suggest that hormonal mechanisms of reproductive tract regulation, considered until recently to be the priority of mammals, have been in place since the era of very early vertebrate evolution, 400 million years before present. These reinforce the view that the endocrine regulatory mechanisms of reproduction in elasmobranchs may serve as models for more highly evolved mammalian species including the human. Indeed, the enormous variety of reproductive tract adaptations in elasmobranchs foreshadows those subsequently evolved in mammals, and because of transitional stages provide needed insight into process which is not possible in the study of mammals alone. © 1993 Wiley‐Liss, Inc.

Endocrine regulation of reproduction in elasmobranchs: Archetype for terrestrial vertebrates

The reproductive biology of the chondrichthyan fishes is remarkably sophisticated. Using both oviparous and viviparous reproductive modes, the group in general has adapted the strategy of producing relatively few young at one time. The oviparous species foreshadow the situation common in oviparous reptiles and universal in birds; viviparous species range from internal incubators to species in which the complexity of placentation and egg yolk reduction approaches the eutherian condition. In certain viviparous elasmobranchs histiotrophic nutrition attains an importance not seen in any other vertebrate group. Associated with the elasmobranch priority on these female reproductive adaptations, a typically vertebrate type of reproductive endocrinology is seen for the first time. This laboratory has focused on ovarian endocrine function and the regulation of the reproductive tract. We report on the steroidogenic capacity of theca, granulosa, and luteal elements and their regulation and correlate this with the natural ovarian cycle and plasma hormone levels. In addition, we describe the biochemistry of the ovarian peptide hormone relaxin and some of its actions on the reproductive tract. As in terrestrial species, relaxin effect on the elasmobranch reproductive tract is steroid hormone-dependent. In general, the results support the concept that the basic endocrine regulatory elements for vertebrate reproductive processes were established in the earliest gnathostomes, the elasmobranchs. These species may therefore provide useful models from which to gain insights into the complexities of higher vertebrate systems that may be obscured by subsequent adaptation to a terrestrial environment and homeothermy.

Pathways for GnRH control of elasmobranch reproductive physiology and behavior

In elasmobranchs, seasonal and developmental changes in reproduction and sexual behavior appear to be regulated by gonadotropin-releasing hormone (GnRH). Circumstantial evidence indicates that GnRH control is primarily via the systemic circulation to pituitary gonadotrophs in the ventral lobe. Additional GnRH effects on other pituitary gonadotrophs and the gonads may also be important. Several forms of GnRH have been reported in sharks and rays; however, the functional significance of the different types is unknown. While immunoreactive-GnRH is present in the preoptic area and hypothalamus, there appears to be little GnRH-staining in the median eminence region. The observations are consistent with a major nonportal distribution of GnRH. Recent studies suggest that the terminal nerve (TN) may be a major source of GnRH. The TN extends from the olfactory sac to the basal telencephalon. It contains abundant immunoreactive-GnRH and has endings in cerebral blood vessels where processes packed with dense-cored vesicles sit adjacent to endothelial cells with active vesicle formation indicative of transcytosis. The TN also projects centrally into the septopreoptic area. The structure may distribute GnRH, both directly to brain nuclei and the olfactory epithelium via synaptic connections and indirectly to brain areas and peripheral reproductive tissues via systemic blood. TN activity may be modulated either through afferent chemical stimulation or via synaptic output from the brain. The results are summarized in a model of suggested neural and endocrine pathways involved in elasmobranch reproduction.

The effect of temperature on steroid biosynthesis by testes of the dogfish, Scyliorhinus caniculus

1. 1. Testes of the dogfish, Scyliorhinus caniculus, were incubated with [ 3H]pregnenolone and [ 3H]testosterone at a range of temperatures from 1 to 36°C. 2. 2. Yields of testosterone from pregnenolone were maximal between 11 and 16°C, corresponding to the breeding temperature of the species. 3. 3. Yields of glucuronides and sulphates increased from 1 to 26°C. 4. 4. Yields of an unidentified polar metabolite increased with temperature and it is suggested that this may play a regulatory role in elasmobranch reproduction.