Population genetics of the fissiparous holothurians Stichopus chloronotus and Holothuria atra (Aspidochirotida): a comparison between the Torres Strait and La Réunion

Collections of about 50 individuals from each of five populations of the fissiparous holothurian species Stichopus chloronotus and four populations of Holothuria atra were made in 1999. These populations were located in the Torres Strait (western Pacific) and La Réunion (western Indian Ocean). Allozyme electrophoretic surveys of five (S. chloronotus) and six (H. atra) loci were conducted to compare patterns of asexual reproduction and to investigate connectivity between regions separated by large geographic distances. Deviations from genotype frequencies expected under Hardy-Weinberg equilibrium, mostly heterozygote excesses, were observed in all populations of both species. The maximum contribution of sexual reproduction (calculated as the maximum number of sexually produced individuals: sample size=N*/N_i) was similar for all S. chloronotus (58-64%) and H. atra (76-92%) populations, and on the same level as previously reported for midshelf reefs of the Great Barrier Reef. The higher values in the latter species indicated greater contributions of asexual reproduction to S. chloronotus populations. Variability was strongly reduced in S. chloronotus populations at La Réunion, with only one locus being variable in that population. When the dataset was reduced to one representative per multi-locus genotype per population to reduce the effect of asexual reproduction on calculations on gene flow, F_ST values were not significantly different from zero, suggesting high gene flow between these regions. However UPGMA cluster analyses using Rogers' genetic distance, roughly clustered populations by region. In the case of H. atra, pooled populations within each region were significantly different from those of the other region. Thus, although some restrictions in gene flow and greater genetic distances between the regions may exist, those differences are distinctly less than those reported in previous studies on echinoderms over similar geographic scales. Despite the importance of asexual reproduction for the maintenance of local population size, this study also confirmed that the potential for widespread dispersal mediated by sexually produced larvae is large.     

Evidence for broadcast spawning as well as brooding in the scleractinian coral Pocillopora damicornis

The reproductive activity of 88 colonies of Pocillopora damicornis at Rottnest Island, Western Australia, was examined over 12 mo (20 August 1988 to 20 August 1989). Larvae were found close to the three new moons of February, March and April, 1989, but in large numbers only during March; 67% of all corals produced larvae. There was no obvious connection between gametogenesis and the production of brooded larvae; however, those corals which did not produce, larvae also did not produce ova but did produce, sperm, and must be considered male colonies. All colonies which produced ova went on to produce planulae. Spermatogenesis occurred throughout the year, but oogenesis occurred between September 1988 and April 1989. Dramatic declines in the percentage of polyps containing eggs and sperm occurred in early February and early April 1989. On these dates the amount of lipid in the tissue of the corals decreased, which suggests that the decline in the percentage of polyps with sperm and eggs was not due to the gametes being resorbed by the polyps. Therefore, P. damicornis at Rottnest Island appears to be both a brooder and a broadcaster with asexual reproduction through brooded larvae and sexual reproduction through spawning of gametes. This accounts for earlier evidence in the literature that some input from sexual reproduction had been occurring in the populations of P. damicornis at Rottnest Island. There is no direct evidence for spawning, as broadcasting of gametes has not been observed, but the indirect evidence provides a strong argument for the existence of this dual reproduction.     

Short-term effects of environmental factors on the asexual reproduction of Aurelia sp. polyps

ABSTRACT

Aurelia is a jellyfish genus common in coastal and estuarine habitats, that are typically affected by human pressures such as heavy metals contamination. Mercury is one of the most dangerous due to its high toxicity, persistence and bioaccumulation. In this paper, we tested for the first time the independent and combined effects of a level of mercury allowable by European regulation in force and hyposalinity on the asexual reproduction of Aurelia sp. polyps, mirroring a realistic scenario. Both mercury (0.07?µg/L, Maximum Allowable Concentration) and salinity (18 and 23 PSU) and the combination of the two factors stimulated the asexual reproduction increasing the number of polyps and buds during the 41 days of experiments. The increment of the asexual reproduction and the absence of suffering and mortality suggest that levels of mercury below the concentration fixed by the law and hyposalinity conditions could promote jellyfish proliferations, according to the hormesis hypothesis.     

Population genetic structures of the fissiparous seastar Coscinasterias acutispina in the Sea of Japan

The morphological characteristics and the population genetic structures of the fissiparous seastar Coscinasterias acutispina were investigated for eight sites in the Sea of Japan in order to clarify the presence of sexual and asexual reproduction. Morphological observation based on arm length showed that fission was common at all eight sites examined, indicating the likely production of clonal individuals. A random amplified polymorphic DNA (RAPD) marker was used to detect clones arising by fission and to assess gene flow among sites. A simulation approach using RAPD data revealed the presence of clonal individuals at almost all sites, suggesting the existence of asexual reproduction. The result of phylogenetic analysis according to RAPD genotype showed no relationship between genetic and geographic distances. Considering the limited movement ability of seastar species during the adult phase, these observations suggest the existence of marked gene flow among sites, due to dispersal of planktonic larvae produced by sexual reproduction. These observations suggest that multi-locus genotypic compositions depend on the relative amounts of recruitment from sexual and asexual reproduction in each population.     

Chaotic genetic patchiness and high relatedness of a poecilogonous polychaete in a heterogeneous estuarine landscape

The genetic structure of benthic marine invertebrates is often described as “chaotic” when genetic structure cannot be explained and barriers to dispersal and gene flow cannot be identified. Here, chaotic patterns of genetic structure for the polychaete Pygospio elegans (Claparède) sampled at 16 locations from the heterogeneous Isefjord–Roskilde Fjord estuary complex in Denmark were found. There was no isolation by distance, and the geography of the estuary complex did not seem to pose a barrier to dispersal and gene flow in this species. We investigated whether characteristics of the environment could be related to the genetic structure and possibly restrict gene flow in this species. Additionally, since P. elegans is poecilogonous, producing larvae with different pelagic developmental periods, we investigated whether observed developmental modes in the samples might clarify the genetic patterns. None of the tested factors explained the population genetic structure. However, a high degree of relatedness among individuals in almost all samples was found. Samples with a larger percentage of young individuals had more related individuals, suggesting that different cohorts could be comprised of individuals with different degrees of relatedness. Relatedness within a site could be increased by limited larval dispersal, collective dispersal of related larvae, sweepstakes reproductive success, or asexual reproduction, but distinguishing between these requires further study. Using a “seascape genetics” approach allowed us to investigate some of the numerous potential factors that could influence population genetic structure in a poecilogonous species.     

Genetic population structure of the seagrass Thalassodendron ciliatum in sandy and rocky habitats in southern Mozambique

The genetic diversity of the dioecious seagrass Thalassodendron ciliatum in six populations in southern Mozambique was studied by means of random amplified polymorphic DNA (RAPD). Samples were taken from sandy and rocky habitats over a span of 880 km. Analysis of molecular variance (AMOVA) showed that most of the genetic variability (71.6% of total genetic variation) was observed within populations. There were also significant genetic differences among populations within each habitat (sandy and rocky). We did not find any significant overall genetic difference between sandy versus rocky populations, indicating that the morphological differences between plants from these two habitats are not maintained by reproductive isolation. There was no significant correlation between geographical and genetic distance, which is discussed in the light of current patterns along the coast. All sampled populations consisted of several genetically distinct individuals, indicating that sexual reproduction is widespread.     

Complex reproductive cycle inEucheilota paradoxica (Hydrozoa: Leptomedusae): Medusae,polyps and frustules produced from medusa stage

Eucheilota paradoxica Mayer, 1900 is a small leptomedusa unknown in the Mediterranean Sea until 1977. Since then, it has become endemic and occurs in great abundance in autumn in the bay of Villefranche-sur-mer. Since no sexual stages have ever been collected, either from the Atlantic Ocean or from the Mediterranean Sea, we decided to investigate the still unknown life cycle of this species. Specimens were obtained in the bay of Villefranche-sur-mer from plankton hauls between 0 and 50 m depth during October and November 1986. At temperatures >18°C, the medusa reproduces intensively by asexual medusal budding (one young medusa liberated per two days). This asexual reproduction explains the rapid proliferation of the species and the origin of the large population every autumn. In cultures maintained at temperatures <18°C, two other reproduction modes, not yet reported in a leptomedusa, were observed: (1) At 16° to 17°C, frustules bud along the canals; these frustules are dormant, giving rise to a small polyp when the temperature increases once more above 18°C. (2) At temperatures close to 15°C each medusa, in place of the gonads, differentiate into one or two polyps; these polyps develop progressively, concomitant with gradual regression of the mother medusa.     

Prey, feeding rates, and asexual reproduction rates of the introduced oligohaline hydrozoan Moerisia lyonsi

Moerisia lyonsi Boulenger (Hydrozoa) medusae and benthic polyps were found at 0 to 5‰ salinity in the Choptank River subestuary of Chesapeake Bay, USA. This species was introduced to the bay at least 30 years before 1996. Medusae and polyps of M. lyonsi are very small and inconspicuous, and may occur widely, but unnoticed, in oligohaline waters of the Chesapeake Bay system and in other estuaries. Medusae consumed copepod nauplii and adults, but not barnacle nauplii, polychaete and ctenophore larvae or tintinnids, in laboratory experiments. Predation rates on copepods by medusae increased with increasing medusa diameter and prey densities. Feeding rates on copepod nauplii were higher than on adults and showed no saturation over the range of prey densities tested (1 to 64 prey l⁻¹). By contrast, predation on copepod adults was maximum (1 copepod medusa⁻¹ h⁻¹) at 32 and 64 copepods l⁻¹. Unexpectedly, M. lyonsi colonized mesocosms at the Horn Point Laboratory during the spring and summer in 4 years (1994 to 1997), and reached extremely high densities (up to 13.6 medusae l⁻¹). Densities of copepod adults and nauplii were low when medusa densities were high, and estimated predation effects suggested that M. lyonsi predation limited copepod populations in the mesocosms. Polyps of M. lyonsi asexually produced both polyp buds and medusae. Rates of asexual reproduction increased with increasing prey availability, from an average total during a 38 d experiment of 9.5 buds polyp⁻¹ when each polyp was fed 1 copepod d⁻¹, to an average total of 146.7 buds polyp⁻¹ when fed 8 copepods d⁻¹. The maximum daily production measured was 8 polyp buds and 22 medusae polyp⁻¹. The colonizing potential of this hydrozoan is great, given the high rates of asexual reproduction, fairly wide salinity tolerance, and existence of a cyst stage. Received: 29 October 1998 / Accepted: 3 March 1999     

Quinones in cockchafers: additional function of a sex attractant as an antimicrobial agent

Summary. Females of the forest cockchafer, Melolontha hippocastani Fabr. (Coleoptera: Scarabaeidae, Melolonthinae), are known to produce toluquinone and 1,4-benzoquinone. The latter component has been shown in our earlier studies to act as a sex attractant that enhances synergistically the attraction of males towards plant volatiles induced by feeding females. Analyses of whole body extracts from adults and grubs (L3) of M. hippocastani by coupled gas chromatography--mass spectrometry revealed the presence of 1,4-benzoquinone and toluquinone also in males and larvae. Quantitative analyses showed that extracts from females contained significantly more of 1,4-benzoquinone and toluquinone than extracts from males. Remarkably, extracts from grubs contained significantly higher amounts of both compounds than extracts from females. Inhibition assays using Escherichia coli, Saccharomyces cerevisiae, and the entomopathogenic fungi Metarhizium anisopliae and Beauveria brongniartii revealed inhibitory effects of 1,4-benzoquinone and toluquinone against all tested microorganisms. However, the amounts necessary for inhibition of entomopathogenic fungi and S. cerevisiae in the laboratory assay were higher than those found in the extracts from M. hippocastani.     

Development of the escape response in larval walleye pollock (Theragra chalcogramma)

The development of the escape response of walleye pollock (Theragra chalcogramma) larvae from attacks by macrozooplanktonic and small-fish predators was quantified in laboratory experiments. Behavior was recorded using video cameras with silhouette illumination from infrared-emitting diodes and by visual observation. Laboratory-reared larvae of 1, 3, 8, 10, 12, 18, 22, 27, 42 days post-hatching, ranging in size from 4 mm to 10 mm total length, were used in the experiments. Even the youngest larvae were observed to exhibit a fast startle response. The percentage of successful larval escapes from the different predators increased as the larvae developed. Euphausiids (Thysanoessa raschii) and amphipods (Calliopiella pratti) often touched larvae but the larvae were usually able to escape and no successful captures of larvae over 22 days old were observed. Although successful escape from initial attacks by three-spine sticklebacks (Gasterosteus aculeatus) increased ontogenetically, sticklebacks were able to consume most larvae, even of the oldest age group, by repeated attacks. Day-old larvae had the lowest percent of escapes after encounters with jellyfish (Sarsia sp.), but the percentage of escapes increased dramatically for 3-day-old larvae. Escape speeds after an attack also increased with age, and tended to be higher after stickleback attacks and lower after jellyfish attacks. This study revealed that the escape response of larval pollock to attack by predators improves rapidly with development during the early larval stage.     

Panmixia in Pocillopora verrucosa from South Africa

The genetic structure of six local collections of Pocillopora verrrucosa from six coral reefs in KwaZulu-Natal, South Africa, was examined using allozyme electrophoresis. The six separate reefs lie within two different reef complexes. Twenty-two enzymes were screened on five buffer systems, but only five polymorphic loci (Gpi-1, Gdh-1, Lgg-2, Lpp-1, Est-1) could be consistently resolved. No significant differences in allelic frequencies were detected among the six sites. All local collections were genotypically diverse, with evidence of only very limited clonal replication at each site. Indeed, the ratio of observed to expected genotypic diversity (mean Go:Ge=0.64ǂ.05 SD), the ratio of observed number of genotypes to the number of individuals (mean Ng:N=0.65ǂ.04 SE), and deviations from the Hardy-Weinberg equilibrium indicate that sexual reproduction plays a major role in the maintenance of the populations. No genetic differentiation was found either within (FSR=0.026ǂ.003 SE) or between (FRT=0.000ǂ.001 SE) reef complexes. The homogeneity of the gene frequencies across the six reefs strongly supports the assumption that the KwaZulu-Natal reef complexes are highly connected by gene flow (Nem=44). The reefs in the southern and central reef complexes along the northern Maputaland coastline can therefore be considered part of a single population.     

Recruitment in the intertidal limpet Lottia digitalis (Patellogastropoda: Lottiidae) may be driven by settlement cues associated with adult habitat

The ribbed limpet, Lottia digitalis, is found high in rocky intertidal habitat throughout its geographic range. In order to identify likely natural settlement locations for larvae of this species, laboratory-reared larvae were settled onto substrata collected from within and near an adult L. digitalis habitat. Of larvae exposed to rock chiseled from within high-intertidal adult habitat, 31.0% and 23.3% underwent metamorphosis during two separate experiments. Similarly, an unidentified filamentous green alga that was isolated from this rock induced metamorphosis in 26.6% and 8.7% of larvae during additional experiments. In contrast, larvae did not metamorphose upon bare rocks or upon rocks encrusted with a crustose corraline alga (CCA) that were collected from lower intertidal zones, nor did they metamorphose upon the macroalgae Ulva sp., Enteromorpha contorta, Alaria marginata, or Polysiphonia sp. The presence of mucus from adult conspecifics during these experiments did not enhance metamorphosis onto rock taken from adult habitat, but it did induce metamorphosis in an average of 13.5% and 7.0% of larvae introduced to the mid-intertidal bare rock and CCA substrata, respectively. Finally, 38.0% and 34.4% of larvae from two experiments underwent metamorphosis when exposed to the high-intertidal barnacle Pollicipes polymerus. These results suggest that larval settlement, rather than differential post-settlement mortality and/or migration, drives recruitment of L. digitalis in high-intertidal habitats.     

Effects of exposure to toxic and non-toxic dinoflagellates on oxygen consumption and locomotion in stage 1 larvae of the crabs Cancer oregonensis and C. magister

Sublethal effects on larval crabs upon exposure to toxic dinoflagellates were examined in the laboratory in early 1999. Specifically, oxygen consumption rates and geotaxis responses were determined for stage 1 larvae of the crabs Cancer oregonensis (Dana) and C. magister Dana that were exposed to non-toxic (Alexandrium tamarense, strain 115) or toxic (A. fundyense, strain 1719) dinoflagellates or to freshly hatched nauplii of the brine shrimp Artemia sp. In C. oregonensis, larvae exposed to the toxic dinoflagellate showed reduced rates of oxygen consumption compared to those exposed to non-toxic dinoflagellates or brine shrimp nauplii. Larvae exposed to a filtrate of the non-toxic dinoflagellate showed no change in oxygen consumption, but a reduced rate when exposed to filtrate from the toxic alga at densities >5᎒² cells ml^-1. In C. magister, larvae exposed to the non-toxic A. tamarense or the toxic A. fundyense had reduced oxygen consumption rates. Larvae exposed to filtrates of non-toxic and toxic dinoflagellates had no change in oxygen consumption. In geotaxis tests, C. oregonensis larvae exposed for 1 day to the toxic A. fundyense reduced their level of locomotion compared to those exposed to non-toxic A. tamarense or to brine shrimp nauplii. C. magister larvae showed no change in activity after a 1-day exposure to the toxic A. fundyense. After a 4-day exposure to A. fundyense, C. magister larvae had much reduced locomotion. Reduced locomotory activity in larvae exposed to toxic algae is consistent with the changes in oxygen consumption rates. Responding to exposure to toxic algae by reducing locomotion may affect vertical migration in these negatively buoyant crab larvae, resulting in sinking below a toxic alga bloom, at least temporarily.     

Reproductive cycle of two commercial species of sea cucumber (Echinodermata: Holothuroidea) from Caribbean Panama

The reproductive status of the holothuroid species Isostichopus badionotus (Selenka, 1867) and Holothuria mexicana (Ludwig, 1875) was studied over 16 months in Bocas del Toro (Panama), from November 1999 to February 2001. Sexual reproduction was evaluated by the gonad index method, and by histology of gonad development. In addition, population structure was assessed based on sex ratio, minimum reproductive size, and length and weight distributions of males and females. The sex ratio in both species was 1:1, with a unimodal population distribution composed mainly of mature individuals. The minimum reproductive length and weight were 13-20 cm and 150 g, respectively, for both species, although reproductive individuals 10 cm in length were also found. A consistently higher gonad index was observed in H. mexicana, due to a high proportion of mature females and males and high gonad indices in most monthly samples. Gametogenesis and spawning patterns seemed to occur throughout the year, with periods of enhanced activity. Two periods of maximum reproductive activity were tentatively identified: July-November for I. badionotus and February-July for H. mexicana, but neither species had a single, sharply defined annual spawning event. Further work on these exploited holothuroids should examine the relationships between reproduction and environmental factors and between reproductive status and recruitment.     

Identification of tenuis of four French Polynesian Carapini (Carapidae: Teleostei)

Four species of adult Carapini (Carapidae) occur on Polynesian coral reefs: Encheliophis gracilis, Carapus boraborensis, C. homei and C. mourlani. Samples collected in Rangiroa and Moorea allowed us to obtain different tenuis (larvae) during their settlement phases or directly inside their hosts. These were separated into four lots on the basis of a combination of pigmentation, meristic, morphological, dental and otolith (sagittae) features. Comparison of these characters with those of the adults allows, for the first time, taxonomic identification of these tenuis-stage larvae.     

Coral polyp budding is probably promoted by a canalized ratio of two morphometric fields

Phenotypic plasticity is the ability of an organism to alter its developmental modes in response to changes in the environment. A major challenge, however, is to explore the existence of a stabilized "morphometric code", a morphological set of buffering rules used repeatedly by members of any species, regardless of environmental factors. Budding of polyps in reef-building corals, an accretive process of the asexual growth form, is thought to be a plastic developmental characteristic. However, we found that intra-tentacular polyp budding in the coral genus Favia from the Red Sea (6 colonies, 37 polyps during a period of 3 years) is not correlated with polyp size. It is initiated after reaching an allometric threshold ratio of two stereotypic, radially symmetrical morphometric landmarks, consisting of the surface area of the polyp and its perimeter.     

Larval reproduction in the ctenophore Mnemiopsis mccradyi (order Lobata)

Dissogony was observed in normal and experimentally manipulated individuals of Mnemiopsis mccradyi in February–April, 1985. The normal oral-aboral length at which sexual reproduction begins in M. mccradyi is 30±5 mm; however, some larvae, of 1.5 to 2.8 mm (oralaboral length), produce viable gametes. During the present study, not all individuals from a particular spawning which were kept in the same conditions became reproductive at an early age. The age at which larval reproduction began varied between individuals. The length of time over which individuals spawned varied considerably — some spawned only once, while others spawned repeatedly. Only the gonads located under the adesophageal ctene rows (and not those associated with the adtentacular rows) produced gametes during the early reproductive period. After larvae stopped spawning, the gonads regressed and the larvae grew to large lobate stages. Individuals which became sexually reproductive as larvae also became sexually reproductive during lobate stages.     

Temperature,salinity, and prey effects on polyp versus medusa bud production by the invasive hydrozoan<Emphasis Type="Italic"> Moerisia lyonsi</Emphasis>

Hydrozoan species are renowned for flexible asexual reproduction, which may predispose them to be successful invaders. Polyps of the invasive hydrozoan Moerisia lyonsi (Boulenger, 1908) have very high rates of asexual production of both polyp and medusa buds. In order to determine how environmental factors affect asexual reproduction in M. lyonsi, the quantitative relationships between polyp bud and medusa bud production were studied in a 31-day laboratory experiment during August 2001. The combined effects of prey (4, 8, 12, 16 Acartia tonsa copepods polyp^–1 day^–1), temperature (20°C, 29°C), and salinity (5, 15, 25) were tested on the development times for polyp buds (DT_p) and medusa buds (DT_m), the total asexual reproduction rate (ARR, no. buds polyp^–1 day^–1), and the ratio of medusa bud to total bud production (R_m). Greater food consumption significantly and directly enhanced ARR and R_m and shortened DT_p and DT_m. A lower temperature (20°C) and higher salinity (25) reduced food consumption, lengthened development times, and decreased ARR and R_m, with opposite effects for the higher temperature (29°C) and lowest salinity (5). The patterns of variation of these reproductive parameters are more complex. DT_m was most sensitive and was significantly and directly affected by all three measured factors. In addition to food consumption, direct effects were seen by temperature on DT_p and by salinity on R_m. ARR was directly affected only by food consumption. Overall, DT_p, DT_m, and R_m were more sensitive to environmental differences than was ARR. More favorable conditions enhanced medusa bud production. The adaptive reproductive processes and their significance for the maintenance and dispersal of M. lyonsi are discussed.Communicated by J.P. Grassle, New Brunswick     

Asexual repooduction in Glossobalanus crozieri (Ptychoderidae,Enteropneusta, Hemichordata)

Several aspects of asexual reproduction in Glossobalanus crozieri (a species first found in Bermuda, but occurring also on the coast of Brazil) have been observed. The results are compared with those by earlier authors on other species of Enteropneusta. A revision of asexual reproduction in the Enteropneusta (Hemichordata) is made. Developmental rates and regeneration were studied in branchiogenital individuals, hepatic individuals and regenerands. The habitat where these forms are found is described. Possible meaning of gonad maturation and sexual reproduction in relation to asexual reproduction in different species is discussed. The need for further studies on the variation of forms which reproduce both sexually and asexually is emphasized, with a view to taxonomic problems, based both upon the proportions of the different body regions and on the number of branchial pores. The present findings indicate that, when compared to the results reported for other species, asexual reproduction in all ptychoderids (Balanoglossus, Glossobalanus and Ptychodera) follows the same general pattern. The phenomenon of asexual reproduction in this group is also confirmed to be worldwide.     

Egg production in the arrow squid Nototodarus gouldi (Cephalopoda: Ommastrephidae), fast and furious or slow and steady?

The process of reproductive maturation and egg release was examined in the temperate shelf squid Nototodarus gouldi. The energy allocation between somatic and reproductive growth from juvenile to mature adult was investigated throughout the life span to determine the underlying energetic strategy adopted by individuals. The relative weight of the mantle, fin and digestive gland remained unchanged during ovarian development, with no significant correlations found between the mantle length (ML)-gonad residuals and the ML-mantle (r=0.01, P>0.05), ML-fin (r=0.07, P>0.05) and ML-digestive gland (r=0.07, P>0.05) residuals. This suggested that energy was not being diverted away from somatic growth during sexual development, and consequently neither muscle nor digestive gland was being utilised as an energy store. Since squid in all maturity stages contained some food in their stomachs (e.g. 66.7% of mature animals), it is likely that the cost of maturation in this species is largely being met by food intake. The energy investment in reproductive tissues was relatively low (mean gonado-somatic index for mature individuals was 9.29% - 0.40%), indicating that only small amounts of energy were being allocated to reproduction at anyone point in time, which is characteristic of a multiple-spawning strategy. Furthermore, oviduct weight was not correlated with body size (r=0.256, P>0.05), suggesting that eggs are not stored for a single release. In all except one individual, ovary weight was consistently heavier than oviduct weight, suggesting that the ovary is not being depleted of oocytes as mature ova move into the oviducts. Additionally, the ovaries of mature females contained a range of oocyte sizes with discrete peaks, indicating a continued production and development of oocyte cohorts. The presence of some individuals with stretched empty oviducts is further evidence that the reproductive strategy of N. gouldi is slow and steady, with eggs possibly being released in discrete batches over a period of time.