Thesis--Auburn University, 2008. / Abstract. Vita. Includes bibliographical references.

Thesis (M.S.)--Montana State University--Bozeman, 2004. / Typescript. Chairperson, Graduate Committee: Alexander V. Zale. Includes bibliographical references (leaves 54-60).

Ocean acidification (OA) together with other anthropogenic perturbations is projected to dramatically alter marine environments over the coming centuries. The vast majority of marine species reproduce by freely spawning sperm directly into the water column, where fertilisation can then either be external or a female can draw sperm into a burrow, brooding chamber or onto her external surface. Hence, sperm are now being released into rapidly changing seawater conditions. In this thesis, I firstly assess what is currently known on the potential for OA and other anthropogenic stressors to influence freely spawned sperm in marine invertebrate taxa. I then present a series of experimental chapters investigating the influence of OA, as a single stressor or in conjunction with a second stressor, copper, on sperm function, physiology and competitive fertilisation performance in a range of invertebrate taxa. My research demonstrates that sperm are vulnerable to the projected changes in seawater carbonate chemistry under OA, with responses observed at all biological levels from sperm physiology, swimming performance, fertilisation ecology and sperm competitiveness. In a multi-stressor experiment on polychaete gametes and larvae, I provide empirical evidence that changes to seawater pH under OA can alter the susceptibility of early life stages including sperm, to the common coastal pollutant copper. Sperm DNA damage increased by 150 % and larval survivorship was reduced by 44 % in combined exposures, than when exposed to copper alone. As a single stressor OA also acted to significantly reduce Arenicola marina sperm swimming speeds and fertilisation success. This work was followed up with a mechanistic investigation of A. marina sperm swimming performance under OA conditions. I found that the length of time between spawning and fertilisation can strongly influence the impact of OA on sperm performance. Key fitness-related aspects of sperm functioning declined after several hours under OA conditions, and these declines could not be explained by changes in sperm ATP content, oxygen consumption or viability. In a final set of experiments, I ran a set of paired competitive fertilisation trials in the sea urchin, Paracentrotus lividus. In addition to reducing fundamental sperm performance parameters, OA conditions affected competitive interactions between males during fertilisation, with potential implications for the proportion of offspring contributed by each male under the new conditions. This work suggests that the ‘best’ males currently may not be the most competitive under OA. Overall this body of work reveals a series of significant changes to sperm performance under OA that might act to perturb sperm functioning in future oceans.

Semiartificial spawning does in fishbastion Cejkovice in the area Ostrov. In the year 2006 was count the produktivity hen fish in a pond 7. After incubation was monitored intensity growth rate fingerling in fingerling pond, next species and size variability natural food and her effect in growth rate. Were writed basic physical and chemistry waters (temperature, pH, content O2). Owing to absence of waters and absence of size fit zooplankton was hunted very little of pikeperch fingerling.

Sexual reproduction in Porites divaricata (Le Sueur, 1820) was studied over a one year period (August, 2012 to August, 2013) in a shallow water (~2m depth) carbonate bank (also described as marginal) habitat at Rodriguez Key, a small island located 1.44 km off the east coast of Key Largo. Porites divaricata is a brooding, gonochoric species with peak reproductive output (planula release) occurring in March and at least a smaller event in May. This species showed an overall female to male sex ratio of 3.5:1. However, during peak reproduction, the sex ratio was 1.2:1. Spawning (sperm release) most likely occurred in late February when 78% of all oocytes and 84% of all spermaries were in stage IV of development. The largest output of stage IV eggs (n=164) occurred at this time, and these oocytes also had the second largest average individual oocyte volume (3.34x10-3 mm3, n = 79) compared to those from other sample dates. The largest average egg volume (3.79x10-3 mm3, n = 8) occurred in May, but with fewer eggs present (n=13). Population surveys showed P. divaricata densities of 7.4 and 17.7 colonies m-² at Site 1 and 2, respectively. Temperature data documented that SSTs were highly variable in the seagrass/coral habitat at site 2 (primary collection site), ranging from 13.1 to 34.9°C, with an average temperature of 25.9°C. Corals at Rodriquez Key are exposed to temperatures much higher (+3.8°C) during the summer and much lower (-8.2°C) in winter compared to corals living at a nearby bank reef (Molasses Reef). This is the first comprehensive reproductive study performed on Porites divaricata and may provide valuable information to the ongoing debate concerning the taxonomic relationships among P. divaricata, P. porites, and P. furcata.

Population dynamics of many intertidal organisms are highly influenced by the abundance and distribution of planktonic larvae in the water column and their arrival on the shore. The brown mussel, Perna perna was used to investigate two of the primary processes that affect population size and dynamics, larval availability and settlement, on the south coast of South Africa. Perna perna is a dominant species on rocky shores of the southern and eastern coasts of South Africa. It creates three-dimensional beds that provide habitats for many other species and hence promotes biodiversity. Larval availability and settlement were examined at different spatial and temporal scales using a nested experimental design. To detect possible relationships between larval availability and settlement, the studies were simultaneous. Two sites, 4km apart, were chosen to investigate mussel settlement patterns. Within each site, three locations (300m from each other) were selected. At each location, five artificial settler collectors were placed at approximately 20cm intervals. Collectors were replaced at a range of time intervals, from daily to seasonal, for 16 months. Each intertidal location was paired with an offshore station, 500m from the shore, where larval availability was measured. At each offshore station, three vertical hauls were collected twice a month using a plankton net. Plankton sampling lasted for 14 months and was designed to examine variability on three temporal scales: seasonal, lunar and daily. The results showed no correlation between the distribution of larvae in the water and settlers on the shore. While larvae were abundant in the water at the start of sampling, they became very rare throughout the rest of the study at both sites and all locations. In contrast, distinct peaks of settler abundance were observed during the seasonal settlement study. In addition to the expected, strong temporal variation that emerged from both studies at all time scales, spatial patterns of variability were also observed. While no spatial effect was detected for the larvae in the water column, there was distinct spatial variation in settlement at the location level: some locations always showed higher settlement than others. These results suggest that, on scales of hundreds of meters to kilometers, larval availability and settlement are very unpredictable in time and that differential delivery of larvae occurs from nearshore waters to the shore. Although the effect of the state of the moon (new or full) was not significant in either study, more settlers seemed to arrive on the shore during new moon. Wind direction did not correlate significantly with settlement. However, the dropping of offshore winds and the prevalence of onshore winds, which are characteristic of summer, may be linked to the start of settlement. Nevertheless, further investigations on tidal or lunar cycles and on the influence of wind on surface currents are required to clarify the effects of moon and wind on settlement.

The objectives of the study were to determine whether or not behaviour could be used to predict spawning in Amphiprion akallopisos, and to document the behaviour of this species under various environmental conditions in captivity. The spawning behaviour of A. akallopisos was studied and quantified. Three behaviour patterns - belly touching, nest cleaning by the female and mutual nest cleaning (by the male and the female) were identified as predictors for spawning. The reproductive behaviour of A.akallopisos under three photoperiods was investigated. The photoperiods were: 14L:10D, 10hr15minL:13hr45minD and a natural photoperiod cycle condensed into three months. A. akallopisos maintained under 14 L:10D exhibited a significantly higher frequency and duration of chasing, nest cleaning and total interaction compared to fish kept under 10hr15minL:13hr45minD. A photoperiod of 14L:10D was selected for further studies. As manipulation of photoperiod did not induce spawning, GnRHa was administered to the fish in the diet at levels of 10, 20, 40 and 80 μg/kg BW. Control groups were fed untreated food. None of the dosages were successful in inducing spawning, or spawning behaviour in A. akallopisos. It is possible that the method of hormone application was not suitable for this species. It is also possible that behaviour may regulate blood hormone levels as opposed to endocrine status influencing behaviour. In the third trial, A. akallopisos was maintained with, and without sea anemones in order to determine whether or not anemone hosts are necessary for spawning. The presence of anemones did not induce spawning and A. akallopisos kept without anemones exhibited significantly more interactive behaviour than fish kept with hosts. Spawning of A. akallopisos at a later date without anemones suggests that anemones are not necessary for spawning in A. akallopisos. Light intensity was investigated as a cue for spawning. The reproductive behaviour of A. akallopisos was then studied under light intensities of 4.16 x 10¹⁵ quanta.sec⁻¹.cm⁻², 8.85 x 10¹⁵ quanta.sec⁻¹.cm⁻² and this intensity plus natural light. Spawning occurred mainly under 8.85 x 10¹⁵ quanta.sec⁻¹.cm⁻². Fish maintained under this light intensity exhibited significantly more of nest cleaning behaviour in terms of frequency and duration than fish maintained under low light intensity. A minimum light intensity of 8.85 x 10¹⁵ quanta. sec⁻¹.cm⁻² is recommended for conditioning of this species. This study can be of practical relevance to hatchery managers who can use the methods developed and record predictors for spawning in A. akallopisos and other Amphiprion species. In this way imminence of spawning may be estimated. Most importantly, the study also has academic merit as little work has been undertaken in this field. Although the observational method used in this study was adequate for the purposes of the investigation, future work of this nature should incorporate other methods of documenting gonadal development such as gonadal staging and GSI in order to obtain more conclusive results.

The recent decline and disappearance of many amphibians around the world is thought to be the sign of an impending sixth mass extinction that is driven by disease, habitat loss and pollution. Reproductive technologies are now required to establish captive colonies followed by reintroduction into suitable habitats. The AMPHIPLEX method is a hormone mixture that has successfully stimulated spawning in several amphibians. However, its extensive application requires further experimentation and knowledge regarding the basic neuroendocrine control of reproduction in amphibians. The role of the catecholamine neurotransmitter dopamine in the regulation of spawning and gonadotropin synthesis was investigated using multiple time- and dose-related approaches in the field and laboratory. These end points were explored in two distantly-related frog species: the Northern leopard frog (Lithobates pipiens) and the Western clawed frog (Silurana tropicalis). Northern leopard frogs were injected during the natural breeding season with three doses of a gonadotropin-releasing hormone agonist (GnRH-A) (0.1 μg/g , 0.2 μg/g and 0.4 μg/g) alone and in combination with two doses of the selective dopamine receptor D2 antagonist metoclopramide (MET) (5 μg/g and 10 μg/g). Injected animals were allowed to breed in mesocosms in an outdoor field. Time to amplexus and oviposition were assessed, and egg mass release, incidences of amplexus, egg mass weight, total egg numbers and fertilization rates were measured. The results revealed no statistically significant interaction between GnRH-A and MET on amplexus and oviposition. A series of GnRH-A dose-response spawning studies were conducted in the Western clawed frog. The current findings indicate that partial ovulation, male sexual behavior and fertilization can be induced by 4 μg/g of GnRH-A alone and in combination with 10 μg/g of MET. This represents a first step towards understanding basic neuroendocrine reproductive mechanisms in this species. These spawning results were paired with a second end point which explored the molecular mechanisms of gonadotropin synthesis in response to GnRH-A and MET alone and in combination. Pituitary gene expression results in the Northern leopard frog indicate a potentiating action of MET when combined with GnRH-A on the mRNA levels of gonadotropin subunits 36 hours following injection. The postulated mechanisms of action are through the upregulation of gonadotropin-releasing hormone receptor 1 and the downregulation of dopamine receptor D2. Such gene expression pathways were similarly explored in the Western clawed frog, however no significant changes in pituitary gonadotropin and receptor gene expression were present at 12 hours post-injection. The hypothesized inhibitory action of dopamine was supported by pituitary gene expression analysis, but not by spawning outcome. The results from this study provide a fundamental framework for future time- and dose-response investigations to improve current spawning methods in amphibians.

Bonefish (Albula spp.) support an economically important catch-and-release recreational fishery, as well as artisanal harvesting, in The Bahamas. Little is known about the large-scale movement patterns of bonefish, yet such information is essential for proper species conservation and management. I used acoustic telemetry to determine large-scale movement patterns of bonefish around Andros, Bahamas, in conjunction with presumed spawning migrations. I conclude that bonefish travel long distances from shallow flats to pre-spawning aggregation sites in proximity to off-shore reef locations. Off-shore movement to deeper reef locations occurs around both new and full moons. This study has also confirmed anecdotal reports that the North Bight is an important spawning migration corridor for bonefish. This information is critical for the protection of bonefish and identifies important habitats (e.g. migration corridors and pre-spawning aggregations) on Andros that warrant protection from coastal degradation or fishing pressures.

Early work on the reproductive seasonality of corals in the Red Sea suggested that corals exhibit temporal reproductive isolation, unlike on the Great Barrier Reef where many species spawn in synchrony. More recent work has however shown high synchrony in the maturity of gametes in Acropora species, suggesting multi-specific spawning is likely to occur in the Red Sea. In this thesis I investigate the patterns of coral reproduction in the central Red Sea. The spawning season in the central Red Sea lasts four months, from April to July and spawning occurs on nights around the full moon. During this period Acropora species show a peak of spawning in April, with some species spawning again in May. The level of synchrony, quantified with a spawning synchrony index, is comparable to other locations where multi-specific spawning has been reported. Observations over two consecutive years show that the synchrony of spawning was lower in spring 2012 than in spring 2011, and thus that spawning patterns are variable from one year to the other. Coral settlement patterns on artificial substrata confirmed a main spawning season in the spring but also supported reproductive data suggesting that some Porites spawn in October-November. Settlement was studied over 2.5 years on a reef, which had suffered recently from high mortality after a local bleaching event. Settlement appeared low but post-bleaching studies from other locations indicated similar abundances and showed that recruits generally did not increase until 5 years after the bleaching event. Abundance of juvenile corals however started to increase significantly three years after the bleaching. Successful recruitment, although low suggests that the coral assemblage on the affected reef will most likely recover as long as it is not affected by another disturbance.