Zoospore Densities Research Articles

Kelp nursery optimisation: density-dependent effects in early life-cycle stages of Ecklonia radiata (Laminariales)

Abstract Kelp aquaculture is an emerging industry outside of Asia. To be successful, this industry requires a reliable production of seedstock, the optimisation of which greatly benefits from a detailed physiological understanding of the microscopic life-cycle stages of the cultured species. This study investigated the impact of six zoospore densities (10–278 mm−2) on the subsequent development of Ecklonia radiata gametophytes and sporophytes. The results showed that germination rates and sex ratio were unaffected by initial zoospore density, but there were significant effects on gametophyte size and sporophyte production. After two weeks, female gametophytes were largest at an initial zoospore density of 40 mm−2 while male gametophytes grew largest at densities below 40 mm−2, but after four weeks gametophyte size showed a negative relationship with initial zoospore density. Significantly more sporophytes developed at initial zoospore densities below 40 individuals mm−2 and no sporophytes were observed at the highest density (271 zoospores mm−2). These results clearly show the importance of initial zoospore density in optimising the nursery stage of kelp aquaculture.

Impact of heavy metal exposure on biological control of a deadly amphibian pathogen by zooplankton

Despite devastating effects on global biodiversity, efficient mitigation strategies against amphibian chytridiomycosis are lacking. Since the free-living pathogenic zoospores of Batrachochytrium dendrobatidis (Bd), the infective stage of this disease, can serve as a nutritious food source for components of zooplankton communities, these groups may act as biological control agents by eliminating zoospores from the aquatic environment. Such pathogen-predator interaction is, however, embedded in the aquatic food web structure and is therefore affected by abiotic factors interfering with these networks. Heavy metals, released from both natural and anthropogenic sources, are widespread contaminants of aquatic ecosystems and may interfere with planktonic communities and thus pathogen elimination rates. We investigated the interaction between zooplankton communities and chytridiomycosis infections in a Flemish agricultural region. Moreover, we also investigated the impact of heavy metal contamination, that was previously investigated in the region and presented in recent work, on zooplankton assemblages and chytridiomycosis infections. Finally, we tested the effect of sublethal concentrations of copper and zinc on Bd removal rates by Daphnia magna in a laboratory assay. Although zinc, copper, nickel and chromium were widely abundant pollutants, heavy metals were no driving force for zooplankton assemblages at our study locations. Moreover, our field survey did not reveal indirect effects of zooplankton assemblages on chytridiomycosis infections. However, sampling occasions testing negative for Bd showed a higher degree of copper contamination compared to positive sampling occasions, indicating a potential inhibitory effect of copper on Bd prevalence. Finally, whereas D. magna significantly reduced zoospore densities in its environment, sublethal concentrations of copper and zinc showed no interference with pathogen removal in the laboratory assay. Our results provide perspectives for further research on such a biological control strategy against chytridiomycosis by optimizing environmental conditions for pathogen predation.

Density-dependence and seasonal variation in reproductive output and sporophyte production in the kelp, Ecklonia radiata.

The kelp, Ecklonia radiata, is an abundant subtidal ecosystem engineer in southern Australia. Density-dependent changes in the abiotic environment engineered by Ecklonia may feedback to affect reproduction and subsequent recruitment. Here, we examined: 1) how the reproductive capacity of Ecklonia individuals in the field (zoospores released · mm-2 reproductive tissue) varied with adult density and time, and 2) how the recruitment of microscopic gametophytes and sporophytes was influenced by zoospore density at two times. Zoospore production did not vary with adult density, with only one month out of ten sampled over a 2-y period showing a significant effect of density. However, zoospore production varied hugely over time, being generally highest in mid-autumn and lowest in mid-late summer. There were strong effects of initial zoospore density on gametophyte and sporophyte recruitment with both a minimum and an optimum zoospore density for sporophyte recruitment, but these varied in time. Almost no sporophytes developed when initial zoospore density was <6.5 · mm-2 in spring or <0.5 · mm-2 in winter with optimum densities of 90-355 · mm-2 in spring and 21-261 · mm-2 in winter, which resulted in relatively high recruitment of 4-7sporophytes · mm-2 . Sporophyte recruitment declined at initial zoospore densities >335 · mm-2 in spring and >261 · mm-2 in winter and was zero at very high zoospore densities. These findings suggest that although adult Ecklonia density does not affect per-capita zoospore production, because there is a minimum zoospore density for sporophyte production, a decline in population-level output could feedback to impact recruitment.

Thermal Performance Curves of Multiple Isolates of Batrachochytrium dendrobatidis, a Lethal Pathogen of Amphibians.

Emerging infectious disease is a key factor in the loss of amphibian diversity. In particular, the disease chytridiomycosis has caused severe declines around the world. The lethal fungal pathogen that causes chytridiomycosis, Batrachochytrium dendrobatidis (Bd), has affected amphibians in many different environments. One primary question for researchers grappling with disease-induced losses of amphibian biodiversity is what abiotic factors drive Bd pathogenicity in different environments. To study environmental influences on Bd pathogenicity, we quantified responses of Bd phenotypic traits (e.g., viability, zoospore densities, growth rates, and carrying capacities) over a range of environmental temperatures to generate thermal performance curves. We selected multiple Bd isolates that belong to a single genetic lineage but that were collected across a latitudinal gradient. For the population viability, we found that the isolates had similar thermal optima at 21°C, but there was considerable variation among the isolates in maximum viability at that temperature. Additionally, we found the densities of infectious zoospores varied among isolates across all temperatures. Our results suggest that temperatures across geographic point of origin (latitude) may explain some of the variation in Bd viability through vertical shifts in maximal performance. However, the same pattern was not evident for other reproductive parameters (zoospore densities, growth rates, fecundity), underscoring the importance of measuring multiple traits to understand variation in pathogen responses to environmental conditions. We suggest that variation among Bd genetic variants due to environmental factors may be an important determinant of disease dynamics for amphibians across a range of diverse environments.

Optimizing the initial cultivation stages of kelp Ecklonia radiata for restoration

Restoration of kelp forests typically relies on transplanting sporophylls to new locations and has limited application in regions with low remnant kelp cover. Cultivated kelp requires fewer sporophylls and is a potential alternative and sustainable source of transplants for large‐scale restoration projects. Naturally sourced fertile sporophylls, however, are still required as “seed” stock in cultivation practices, thus optimizing cultivation methods is important to minimize this dependency on wild stocks. To assist in optimizing the early‐stage cultivation methods for restoring beds of the laminarian kelp Ecklonia radiata, we first tested the effects of sporophyll transport, storage, and dehydration on zoospore release. We then tested for effects of inoculum storage temperature on zoospore abundance, and lastly for media sterilization and inoculum concentration effects on both zoospore settlement and resulting gametophyte densities. Our results show, to maximize zoospore release, sporophylls should be transported dry and inoculated within 3 hours. Inoculum can be stored at 4°C without affecting zoospore abundance and should be added to sterilized media at lower concentrations to reduce settled zoospore density and improve post‐settlement survival of the gametophyte stage. This study provides practical recommendations for optimizing the initial cultivation procedures of E. radiata. To develop a full life‐cycle cultivation protocol for kelp restoration purposes, future research should focus on optimizing sporophyte production, out‐planting and transplanting techniques.

The potential of zooplankton in constraining chytrid epidemics in phytoplankton hosts.

Fungal diseases threaten natural and man‐made ecosystems. Chytridiomycota (chytrids) infect a wide host range, including phytoplankton species that form the basis of aquatic food webs and produce roughly half of Earth's oxygen. However, blooms of large or toxic phytoplankton form trophic bottlenecks, as they are inedible to zooplankton. Chytrids infecting inedible phytoplankton provide a trophic link to zooplankton by producing edible zoospores of high nutritional quality. By grazing chytrid zoospores, zooplankton may induce a trophic cascade, as a decreased zoospore density will reduce new infections. Conversely, fewer infections will not produce enough zoospores to sustain long‐term zooplankton growth and reproduction. This intricate balance between zoospore density necessary for zooplankton energetic demands (growth/survival), and the loss in new infections (and thus new zoospores) because of grazing was tested empirically. To this end, we exposed a cyanobacterial host (Planktothrix rubescens) infected by a chytrid (Rizophydium megarrhizum) to a grazer density gradient (the rotifer Keratella cf. cochlearis). Rotifers survived and reproduced on a zoospore diet, but the Keratella population growth was limited by the amount of zoospores provided by chytrid infections, resulting in a situation where zooplankton survived but were restricted in their ability to control disease in the cyanobacterial host. We subsequently developed and parameterized a dynamical food‐chain model using an allometric relationship for clearance rate to assess theoretically the potential of different‐sized zooplankton groups to restrict disease in phytoplankton hosts. Our model suggests that smaller‐sized zooplankton may have a high potential to reduce chytrid infections on inedible phytoplankton. Together, our results point out the complexity of three‐way interactions between hosts, parasites, and grazers and highlight that trophic cascades are not always sustainable and may depend on the grazer's energetic demand.

Guidance of zoospores by potassium gradient sensing mediates aggregation.

The biflagellate zoospores of some phytopathogenic Phytophthora species spontaneously aggregate within minutes in suspension. We show here that Phytophthora parasitica zoospores can form aggregates in response to a K+ gradient with a particular geometric arrangement. Using time-lapse live imaging in macro- and microfluidic devices, we defined (i) spatio-temporal and concentration-scale changes in the gradient, correlated with (ii) the cell distribution and (iii) the metrics of zoospore motion (velocity, trajectory). In droplets, we found that K+-induced aggregates resulted from a single biphasic temporal sequence involving negative chemotaxis followed by bioconvection over a K+ gradient concentration scale [0-17 mM]. Each K+-sensing cell moved into a region in which potassium concentration is below the threshold range of 1-4 mM, resulting in swarming. Once a critical population density had been achieved, the zoospores formed a plume that migrated downward, with fluid advection in its wake and aggregate formation on the support surface. In the microfluidic device, the density of zoospores escaping potassium was similar to that achieved in droplets. We discuss possible sources of K+ gradients in the natural environment (zoospore population, microbiota, plant roots, soil particles), and implications for the events preceding inoculum formation on host plants.

Quantification of laminarialean zoospores in seawater by real‐time PCR

SUMMARYLaminarialean plants undergo heteromorphic alternation of generations between the macroscopic diploid sporophyte and the microscopic haploid gametophyte. The generations change through the formation and release of asexual or sexual reproductive cells. It is difficult to monitor the release and diffusion of zoospores into the environment. Furthermore, even if zoospores can be detected, species identification remains difficult. This study attempted to develop a sensitive and fast identification method for laminarialean zoospores using quantitative PCR. In addition, we aimed to estimate the density of zoospores in natural seawater. Specific primers for Saccharina japonica and Undaria pinnatifida were designed and used to estimate the quantity of zoospores in seawater. DNA samples (environmental DNA) were collected from seawater once or twice each month for 2 years at the same area, and seasonal variations in the release of zoospores was monitored. The estimated maturation period based on the number of released zoospores in this study was comparable with those of the previously reported maturation periods of S. japonica and U. pinnatifida sporophytes. This supports the validity of our method in estimating zoospore release from laminarialean plants. The method will be a useful tool for ecological studies on these commercially relevant species.

Leucine Regulates Zoosporic Germination and Infection by Phytophthora erythroseptica.

Pink rot (Phytophthora erythroseptica) of potato is a major concern in many potato production regions. The pathogen produces zoospores that serve as a primary inoculum for infection. To understand how the pink rot incidence is related to pathogen population, qualitative, and quantitative chemical analyses were conducted. It was demonstrated that P. erythroseptica zoospores required a minimal population of 103 zoospores/ml (threshold) for initiating germination and the subsequent infection; the percentage of zoosporic germination was positively correlated with the density of zoospores above the threshold. To elucidate the density-dependent behavior, zoospore exudate (ZE) was extracted from high-density (105/ml) zoospore suspension. Zoosporic inocula of P. erythroseptica at different concentrations were inoculated on potato tubers. Necrotic lesions were caused by inoculum with 100 zoospores per inoculation site; 5 zoospores per site did not cause lesions on the tuber. However, five zoospores did cause lesions when they were placed in ZE, suggesting ZE contained chemical compounds that regulate germination of zoospores. ZE was collected and analyzed using liquid chromatography mass spectroscopy (LC-MS). Results showed that the amino acid leucine was associated with zoosporic germination. Therefore, zoosporic germination and infection of P. erythroseptica were mediated by signaling molecules secreted from zoospores.

Marine anti-biofouling efficacy of amphiphilic poly(coacrylate) grafted PDMSe: effect of graft molecular weight

There is currently strong motivation due to ecological concerns to develop effective anti-biofouling coatings that are environmentally benign, durable, and stable for use by the maritime industry. The antifouling (AF) and fouling-release (FR) efficacy of amphiphilic, charged copolymers composed of ~52% acrylamide, ~34% acrylic acid, and ~14% methyl acrylate grafted to poly(dimethyl siloxane) (PDMSe) surfaces were tested against zoospores of the green alga Ulva linza and the diatom Navicula incerta. The biofouling response to molecular weight variation was analyzed for grafts ranging from ~100 to 1,400 kg mol−1, The amphiphilic coatings showed a marked improvement in the FR response, with a 55% increase in the percentage removal of diatoms and increased AF efficacy, with 92% reduction in initial attachment density of zoospores, compared to PDMSe controls. However, graft molecular weight, in the range tested, was statistically insignificant. Grafting copolymers to PDMSe embossed with the Sharklet™ microtopography did not produce enhanced AF efficacy.

Prevalence and dynamics of the zoosporic pathogen Catenaria uncinata in a natural population of the midge Glyptotendipes lobiferus

A qPCR assay specific for zoospores of Catenaria uncinata, a fungal parasite in eggs of the midge Glyptotendipes lobiferus, was developed and used in parallel with traditional microscopic methods in a season-long study of a C. uncinata/G. lobiferus association in a local pond. Twenty-six consecutive weekly collections of egg masses were screened with a microscope to obtain percentages of infection and mortality in organogenetic egg masses and weekly water samples were processed by absolute quantification using qPCR to obtain estimates of zoospore density. Overall, 36.0% of G. lobiferus egg masses were infected to varying degrees and 11.2% of eggs were killed by C. uncinata. Continuous infection of egg masses occurred during a 6-wk period in May–June and a 7-wk period in September–October. Infection by C. uncinata was absent during a 10-week interval between periods of infection. Abrupt declines in zoospore density occurred during both infection periods and occurred only when water temperatures met or exceeded the viability threshold for zoospores (⩾31.0°C). The episodic death of zoospores during weeks in which egg infection and mortality levels were continuous likely resulted from distribution of zoospores throughout the water column and a temperature gradient in which zoospores sampled near the surface were subjected to lethal temperatures while non-sampled zoospores at lower depths were provided low temperature sanctuary. The hiatus of infection during the 10-week interval was likely due to lethal temperatures throughout the water column as average water temperatures exceeded 31.0°C over the period. A positive correlation between weekly zoospore densities obtained from qPCR and levels of infection/mortality in egg masses obtained from counts with a microscope supports the use of the qPCR assay alone in future studies that can rapidly and accurately determine parasite presence, prevalence and geographical range.

Understory algae and low light reduce recruitment of the habitat-forming kelp Ecklonia radiata

Kelp forests form important habitat in temperate subtidal regions but are threatened by a range of stressors that impact their abundance. In this study, we determined the effect of understory algae, a potential competitor of kelp, on the recruitment of the Australasian kelp, Ecklonia radiata, via their role in reducing light availability for early life-cycle stages of Ecklonia. A field experiment manipulating the abundance of understory algae showed no recruitment of outplanted microscopic sporophytes or naturally recruiting macroscopic sporophytes with a high (> 80%) cover of understory algae. In contrast, both microscopic and macroscopic sporophytes recruited into plots with a medium (40 to 60%) and low (< 20%) cover of understory algae. Light appears to be the important mechanism for the inhibition of kelp recruitment because a high cover of understory algae blocks up to 90% of available light for Ecklonia recruits. A laboratory experiment showed that low light (similar to the levels beneath the understory of high-coverage algal canopies) inhibited Ecklonia gametophyte growth and prevented microscopic sporophyte recruitment. Both high light (similar to levels when there is little understory algal canopy) and high zoospore density resulted in high sporophyte recruitment, suggesting important additive effects of those factors. This study indicates that an increase in the cover of understory algae results in low light, which inhibits Ecklonia sporophyte recruitment. The resilience of kelp will be strongly influenced by recruitment, and our results highlight that a decline in kelp canopy cover which leads to an increase in understory algae can ultimately feedback to negatively affect kelp recruitment.

Development of quantification bioassays for live propagules of Phytophthora sojae in soil

To estimate inoculum density of Phytophthora sojae in natural soils to predict disease risk and to study the ecology of P. sojae in the soil, we developed two bioassays to quantify zoospores and another for oospores of P. sojae in soil based on inoculation of Mollisol (the most conducive soil type for this disease according to our disease survey) with a known density of zoospores or oospores of enhanced green fluorescent protein (EGFP)-labeled P. sojae Eps597-3 (pathotype 1a3c7). A linear regression equation, \( y_{ 1} = 1.0 9 2 7x_{ 1} + 7. 3 6 5 2 \) (Eq. 1), and an index regression equation, \( {\text{y}}_{ 2} \, \;{ = }\; 1 8 6 2. 9 {\text{e}}^{{0.00 9 5 {\text{x}}_{ 2} }} \) (Eq. 2), were established to quantify the zoospores in the soil, where x1 is the concentration of zoospores counted directly using a light microscope and y1 is the concentration of zoospores added to a soil suspension, x2 is the number of zoospores at the margin of 18 leaf discs, and y2 is the number of zoospores added to 10 mL of a soil suspension. Another linear regression equation, \( y_{ 3} = 3. 3 8 3 6x_{ 3} + 20 7.0 5 \) (Eq. 3), was determined to quantify the oospores in the soil, where x3 is the number of zoosporangia at the margin of 18 leaf discs, and y3 is the number of oospores added to 10 mL of a soil suspension. These three equations provided high reproducibility and accuracy. To verify their applicability, two other isolates with different pathotypes (3a3b678 and 1a1d23a3c578) and two other soil types (Albic luvisols and Aquic cambosols) were tested. Pathotype and soil type did not markedly affect the parameters in the three equations, suggesting that these three equations are accurate enough to serve as universal quantification bioassays of live zoospores and oospores of P. sojae isolates of any pathotype in any soil. Equations (1) and (2) can be used only to estimate zoospores, Eq. (2) for zoospores, and Eq. (3) for oospores of P. sojae in natural or inoculated soil. This method can be finished within 2–3 h for Eq. (1), 4–5 h for Eq. (2), and 7–10 days for Eq. (3).

Experimental evolution alters the rate and temporal pattern of population growth in Batrachochytrium dendrobatidis, a lethal fungal pathogen of amphibians.

Virulence of infectious pathogens can be unstable and evolve rapidly depending on the evolutionary dynamics of the organism. Experimental evolution can be used to characterize pathogen evolution, often with the underlying objective of understanding evolution of virulence. We used experimental evolution techniques (serial transfer experiments) to investigate differential growth and virulence of Batrachochytrium dendrobatidis (Bd), a fungal pathogen that causes amphibian chytridiomycosis. We tested two lineages of Bd that were derived from a single cryo-archived isolate; one lineage (P10) was passaged 10 times, whereas the second lineage (P50) was passaged 50 times. We quantified time to zoospore release, maximum zoospore densities, and timing of zoospore activity and then modeled population growth rates. We also conducted exposure experiments with a susceptible amphibian species, the common green tree frog (Litoria caerulea) to test the differential pathogenicity. We found that the P50 lineage had shorter time to zoospore production (Tmin), faster rate of sporangia death (ds), and an overall greater intrinsic population growth rate (λ). These patterns of population growth in vitro corresponded with higher prevalence and intensities of infection in exposed Litoria caerulea, although the differences were not significant. Our results corroborate studies that suggest that Bd may be able to evolve relatively rapidly. Our findings also challenge the general assumption that pathogens will always attenuate in culture because shifts in Bd virulence may depend on laboratory culturing practices. These findings have practical implications for the laboratory maintenance of Bd isolates and underscore the importance of understanding the evolution of virulence in amphibian chytridiomycosis.

In Situ Production of Zoospores by Five Species of Phytophthora in Aqueous Environments for Use as Inocula

The goal of this study was to develop a procedure that could be used to evaluate the potential susceptibility of aquatic plants used in constructed wetlands to species of Phytophthora commonly found in nurseries. V8 agar plugs from actively growing cultures of three or four isolates of Phytophthora cinnamomi, P. citrophthora, P. cryptogea, P. nicotianae, and P. palmivora were used to produce inocula. In a laboratory experiment, plugs were placed in plastic cups and covered with 1.5% nonsterile soil extract solution (SES) for 29 days, and zoospore presence and activity in the solution were monitored at 2- or 3-day intervals with a rhododendron leaf disk baiting bioassay. In a greenhouse experiment, plugs of each species of Phytophthora were placed in plastic pots and covered with either SES or Milli-Q water for 13 days during both summer and winter months, and zoospore presence in the solutions were monitored at 3-day intervals with the baiting bioassay and by filtration. Zoospores were present in solutions throughout the 29-day and 13-day experimental periods but consistency of zoospore release varied by species. In the laboratory experiment, colonization of leaf baits decreased over time for some species and often varied among isolates within a species. In the greenhouse experiment, bait colonization decreased over time in both summer and winter, varied among species of Phytophthora in the winter, and was better in Milli-Q water. Zoospore densities in solutions were greater in the summer than in the winter. Decreased zoospore activities for some species of Phytophthora were associated with prolonged temperatures below 13 or above 30°C in the greenhouse. Zoospores from plugs were released consistently in aqueous solutions for at least 13 days. This procedure can be used to provide in situ inocula for the five species of Phytophthora used in this study so that aquatic plant species can be evaluated for potential susceptibility.

A fungal pathogen of amphibians, Batrachochytrium dendrobatidis, attenuates in pathogenicity with in vitro passages.

Laboratory investigations into the amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), have accelerated recently, given the pathogen’s role in causing the global decline and extinction of amphibians. Studies in which host animals were exposed to Bd have largely assumed that lab-maintained pathogen cultures retained the infective and pathogenic properties of wild isolates. Attenuated pathogenicity is common in artificially maintained cultures of other pathogenic fungi, but to date, it is unknown whether, and to what degree, Bd might change in culture. We compared zoospore production over time in two samples of a single Bd isolate having different passage histories: one maintained in artificial media for more than six years (JEL427-P39), and one recently thawed from cryopreserved stock (JEL427-P9). In a common garden experiment, we then exposed two different amphibian species, Eleutherodactylus coqui and Atelopus zeteki, to both cultures to test whether Bd attenuates in pathogenicity with in vitro passages. The culture with the shorter passage history, JEL427-P9, had significantly greater zoospore densities over time compared to JEL427-P39. This difference in zoospore production was associated with a difference in pathogenicity for a susceptible amphibian species, indicating that fecundity may be an important virulence factor for Bd. In the 130-day experiment, Atelopus zeteki frogs exposed to the JEL427-P9 culture experienced higher average infection intensity and 100% mortality, compared with 60% mortality for frogs exposed to JEL427-P39. This effect was not observed with Eleutherodactylus coqui, which was able to clear infection. We hypothesize that the differences in phenotypic performance observed with Atelopus zeteki are rooted in changes of the Bd genome. Future investigations enabled by this study will focus on the underlying mechanisms of Bd pathogenicity.

Environmental influences on kelp performance across the reproductive period: an ecological trade-off between gametophyte survival and growth?

Most kelps (order Laminariales) exhibit distinct temporal patterns in zoospore production, gametogenesis and gametophyte reproduction. Natural fluctuations in ambient environmental conditions influence the intrinsic characteristics of gametes, which define their ability to tolerate varied conditions. The aim of this work was to document seasonal patterns in reproduction and gametophyte growth and survival of Ecklonia radiata (C. Agardh) J. Agardh in south-western Australia. These results were related to patterns in local environmental conditions in an attempt to ascertain which factors explain variation throughout the season. E. radiata was fertile (produced zoospores) for three and a half months over summer and autumn. Every two weeks during this time, gametophytes were grown in a range of temperatures (16–22°C) in the laboratory. Zoospore densities were highly variable among sample periods; however, zoospores released early in the season produced gametophytes which had greater rates of growth and survival, and these rates declined towards the end of the reproductive season. Growth rates of gametophytes were positively related to day length, with the fastest growing recruits released when the days were longest. Gametophytes consistently survived best in the lowest temperature (16°C), yet exhibited optimum growth in higher culture temperatures (20–22°C). These results suggest that E. radiata releases gametes when conditions are favourable for growth, and E. radiata gametophytes are tolerant of the range of temperatures observed at this location. E. radiata releases the healthiest gametophytes when day length and temperature conditions are optimal for better germination, growth, and sporophyte production, perhaps as a mechanism to help compete against other species for space and other resources.

Reproductive synchrony in a habitat-forming kelp and its relationship with environmental conditions

Key biological processes such as the timing and synchrony of reproduction, are sensitive to fluctuations in the environment, as well as alterations in the physiology and behaviour of individuals and the ecology of populations. The main aim of this study was to identify patterns in reproductive timing and relationships with prevailing environmental conditions, for the major habitat-forming alga in temperate Australia, Ecklonia radiata (C. Agardh) J. Agardh. We observed strong synchronous patterns of zoospore production across spatial scales from hundreds of metres to tens of kilometres, with populations of E. radiata reproducing from mid-summer to the end of autumn. High zoospore densities were positively related to sea temperature and E. radiata released zoospores in temperatures well above previously documented thermal tolerance limits for other Laminariales. Reproductive timing was also negatively related to seasonal increase in wave exposure. We conclude that E. radiata undergoes synchronous reproduction which can be linked to its annual growth cycle and natural environmental fluctuations, to promote growth and survival of recruits.

Engineered antifouling microtopographies: kinetic analysis of the attachment of zoospores of the green alga Ulva to silicone elastomers

Microtopography has been demonstrated as an effective deterrent to biofouling. The majority of published studies are fixed-time assays that raise questions regarding the kinetics of the attachment process. This study investigated the time-dependent attachment density of zoospores of Ulva, in a laboratory assay, on a micropatterned and smooth silicone elastomer. The attachment density of zoospores was reduced on average 70–80% by the microtopography relative to smooth surfaces over a 4 h exposure. Mapping the zoospore locations on the topography revealed that they settled preferentially in specific, recessed areas of the pattern. The kinetic data fit, with high correlation (r 2 > 0.9), models commonly used to describe the adhesion of bacteria to surfaces. The grouping of spores on the microtopography indicated that the pattern inhibited the ability of attached spores to recruit neighbors. This study demonstrates that the antifouling mechanism of topographies may involve disruption of the cooperative effects exhibited by fouling organisms such as Ulva.

Vertical distribution of kelp zoospores

Cie D.K. and Edwards M.S. 2011. Vertical distribution of kelp zoospores. Phycologia 50: 340–350. DOI: 10.2216/10-48.1The widely accepted theory of kelp propagule dispersal is that reproductive zoospores are released near the benthos and passively dispersed horizontally via advective and diffusive transport. However, it is possible that these zoospores can also be dispersed vertically away from the benthos and into greater unidirectional flow, thus increasing their dispersal potential. To investigate this, we developed a vertical spore profiler to collect settled zoospores at 2, 4, 6, 8, and 10 m above the substrate within and adjacent to the Point Loma Kelp Forest, California. Concurrently, we deployed a Nortek Aquadopp™ acoustic Doppler current profiler (ADCP) to measure current velocities at different depths within and outside of the forest. Our results indicate that kelp zoospores are vertically dispersed throughout the water column, reaching at least 10 m off the benthos, but that zoospore density exponentially decreases with increasing height off the substrate. Furthermore, large differences in overall zoospore abundance were observed among different areas within and outside of the kelp forest and among different times of the year. Variability in zoospore densities coincided with differences in the physical environment (e.g. wave activity, horizontal flow, irradiance). Our ADCP data revealed that alongshore currents increased with increasing distance above the benthos, thereby increasing dispersal potential at these depths. However, exposure of the zoospores to irradiances characteristic of shallower water resulted in reduced settlement and postsettlement competency. Although the majority of released zoospores remained near the benthos, our data support the idea that vertical transport of kelp zoospores into shallower portions of the water column, where unidirectional currents are greater, may provide an alternate vector for long-distance dispersal.