Composition and structure of microbial communities from stromatolites of Hamelin Pool in Shark Bay, Western Australia

Stromatolites, organosedimentary structures formed by microbial activity, are found throughout the geological record and are important markers of biological history. More conspicuous in the past, stromatolites occur today in a few shallow marine environments, including Hamelin Pool in Shark Bay, Western Australia. Hamelin Pool stromatolites often have been considered contemporary analogs to ancient stromatolites, yet little is known about the microbial communities that build them. We used DNA-based molecular phylogenetic methods that do not require cultivation to study the microbial diversity of an irregular stromatolite and of the surface and interior of a domal stromatolite. To identify the constituents of the stromatolite communities, small subunit rRNA genes were amplified by PCR from community genomic DNA with universal primers, cloned, sequenced, and compared to known rRNA genes. The communities were highly diverse and novel. The average sequence identity of Hamelin Pool sequences compared to the >200,000 known rRNA sequences was only approximately 92%. Clone libraries were approximately 90% bacterial and approximately 10% archaeal, and eucaryotic rRNA genes were not detected in the libraries. The most abundant sequences were representative of novel proteobacteria (approximately 28%), planctomycetes ( approximately 17%), and actinobacteria (approximately 14%). Sequences representative of cyanobacteria, long considered to dominate these communities, comprised <5% of clones. Approximately 10% of the sequences were most closely related to those of alpha-proteobacterial anoxygenic phototrophs. These results provide a framework for understanding the kinds of organisms that build contemporary stromatolites, their ecology, and their relevance to stromatolites preserved in the geological record.     

Composition and Structure of Microbial Communities from Stromatolites of Hamelin Pool in Shark Bay, Western Australia

Stromatolites, organosedimentary structures formed by microbial activity, are found throughout the geological record and are important markers of biological history. More conspicuous in the past, stromatolites occur today in a few shallow marine environments, including Hamelin Pool in Shark Bay, Western Australia. Hamelin Pool stromatolites often have been considered contemporary analogs to ancient stromatolites, yet little is known about the microbial communities that build them. We used DNA-based molecular phylogenetic methods that do not require cultivation to study the microbial diversity of an irregular stromatolite and of the surface and interior of a domal stromatolite. To identify the constituents of the stromatolite communities, small subunit rRNA genes were amplified by PCR from community genomic DNA with universal primers, cloned, sequenced, and compared to known rRNA genes. The communities were highly diverse and novel. The average sequence identity of Hamelin Pool sequences compared to the >200,000 known rRNA sequences was only ~92%. Clone libraries were ~90% bacterial and ~10% archaeal, and eucaryotic rRNA genes were not detected in the libraries. The most abundant sequences were representative of novel proteobacteria (~28%), planctomycetes (~17%), and actinobacteria (~14%). Sequences representative of cyanobacteria, long considered to dominate these communities, comprised <5% of clones. Approximately 10% of the sequences were most closely related to those of α-proteobacterial anoxygenic phototrophs. These results provide a framework for understanding the kinds of organisms that build contemporary stromatolites, their ecology, and their relevance to stromatolites preserved in the geological record.     

Microbial diversity of extant stromatolites in the hypersaline marine environment of Shark Bay, Australia

Stromatolites have been present on Earth, at various levels of distribution and diversity, for more than 3 billion years. Today, the best examples of stromatolites forming in hypersaline marine environments are in Hamelin Pool at Shark Bay, Western Australia. Despite their evolutionary significance, little is known about their associated microbial communities. Using a polyphasic approach of culture-dependent and culture-independent methods, we report the discovery of a wide range of microorganisms associated with these biosedimentary structures. There are no comparable reports combining these methodologies in the survey of cyanobacteria, bacteria, and archaea in marine stromatolites. The community was characterized by organisms of the cyanobacterial genera Synechococcus, Xenococcus, Microcoleus, Leptolyngbya, Plectonema, Symploca, Cyanothece, Pleurocapsa and Nostoc. We also report the discovery of potentially free-living Prochloron. The other eubacterial isolates and clones clustered into seven phylogenetic groups: OP9, OP10, Marine A group, Proteobacteria, Low G+C Gram-positive, Planctomycetes and Acidobacteria. We also demonstrate the presence of sequences corresponding to members of halophilic archaea of the divisions Euryarchaeota and Crenarchaeota and methanogenic archaea of the order Methanosarcinales. This is the first report of such archaeal diversity from this environment. This study provides a better understanding of the microbial community associated with these living rocks.     

Two new opecoelids (Digenea) in the flatfish Pseudorhombus jenynsi (Bleeker) from Shark Bay,Western Australia

The first marine fish digeneans recorded from Shark Bay, Western Australia are described from the flatfish Pseudorhombus jenynsi. Macvicaria dampieri n. sp. differs from the other members of the genus in its undivided, straight, saccular seminal vesicle, and the definition of the genus is extended to include this feature. The following new combinations are made: Macvicaria branchiostegi (Yamaguti, 1937) [originally Plagioporus] and M. cynoglossi (Madhavi, 1975) [originally P.]. Cainocreadium alanwilliamsi n. sp. is distinguished by the variable anterior extent of its excretory vesicle, which reaches to a level ranging between the posterior margin of the ventral sucker and just into the forebody, and by its relatively short forebody.     

Northern range extension of the seagrasses Halophila johnsonii and Halophila decipiens along the east coast of Florida,USA

The northern geographic limit for Halophila johnsonii and Halophila decipiens has been reported as Sebastian Inlet, within the Indian River Lagoon, Florida. Surveys conducted in August 2007 determined the new northern limit to be 21.5 km north of the previously known limit. This new northern limit is a 10% range extension for H. johnsonii, a federally threatened species. We conclude that these range extensions are recent, based on (1) the small size of patches; (2) unusually good water clarity conditions due to a recent drought; (3) recent mild/warmer winters; and (4) a recent mechanism for transporting propagules, the numerous hurricanes of 2004. Although this recent range extension is considered ephemeral, similar range extensions may have occurred in the past and may occur again in the future under favorable conditions given the high capacity of these two species for dispersal to favorable sites. The northern limits of these species should not be viewed as static locations; rather, they must be considered dynamic features.     

Megagametogenesis in Halophila johnsonii, a threatened seagrass with no known seeds,and the seed-producing Halophila decipiens (Hydrocharitaceae)

Megagametogenesis, the development of a megaspore into an embryo sac, has been identified in the seagrass Halophila johnsonii, a threatened species with no known sexual reproduction or seeds. Megagametogenesis in H. johnsonii was compared with megagametophyte development in Halophila decipiens, a related species known to readily produce viable seeds. In both species, ovules were structurally similar, megaspore mother cells were seen in premeiotic ovules, and linear tetrads and megagametophytes with two to eight nuclei were present in postmeiotic ovules. However, H. decipiens postmeiotic ovules had a chalazal pouch that was absent in the postmeiotic ovules of H. johnsonii. Late-stage H. decipiens ovules also contained embryos, indicating that they had been fertilized, whereas all late-stage H. johnsonii ovules were degrading and showed no signs of fertilization. These observations suggest that meiosis does occur in H. johnsonii megasporocytes, leading to the formation of viable megagametophytes and egg cells that could be fertilized if pollination occurred. Thus, the lack of seed set is due to a lack of pollination rather than any loss of capacity to produce seeds in this species.     

Correlations between salinity and growth of the seagrass Amphibolis antarctica (labill.) Sonder & Aschers., In Shark Bay,Western Australia,using a new method for measuring production rate

A method of estimating above-ground productivity in situ of the seagrass Amphibolis antarctica (Labill.) Sonder & Aschers. has been devised, using tags to determine rates of leaf turnover. This has proved an effective tool in establishing the behavior of the species in relation to the gradient of increasing salinity which is present in Shark Bay. No seagrass was found beyond 64%_o, but measurement of production and biomass within dense patches of seagrass at different salinities revealed that these were at a maximum at a salinity of 42%_o, decreasing as the salinity increased and also at lower oceanic concentrations. Production rates ranged from 2 to 17 g dry weight m^?2 day^?1 with biomass from 600 to 2000 g m^?2, thus Amphibolis antarctica is one of the more productive Australian seagrass species, even in the hypersaline conditions of the Bay. Despite the obvious correlation between above-ground production and salinity, it is pointed out that the results are not taken to imply causality.     

Active eukaryotes in microbialites from Highborne Cay,Bahamas, and Hamelin Pool (Shark Bay), Australia

Microbialites are organosedimentary structures that are formed through the interaction of benthic microbial communities and sediments and include mineral precipitation. These lithifying microbial mat structures include stromatolites and thrombolites. Exuma Sound in the Bahamas, and Hamelin Pool in Shark Bay, Western Australia, are two locations where significant stands of modern microbialites exist. Although prokaryotic diversity in these structures is reasonably well documented, little is known about the eukaryotic component of these communities and their potential to influence sedimentary fabrics through grazing, binding and burrowing activities. Accordingly, comparisons of eukaryotic communities in modern stromatolitic and thrombolitic mats can potentially provide insight into the coexistence of both laminated and clotted mat structures in close proximity to one another. Here we examine this possibility by comparing eukaryotic diversity based on Sanger and high-throughput pyrosequencing of small subunit ribosomal RNA (18S rRNA) genes. Analyses were based on total RNA extracts as template to minimize input from inactive or deceased organisms. Results identified diverse eukaryotic communities particularly stramenopiles, Alveolata, Metazoa, Amoebozoa and Rhizaria within different mat types at both locations, as well as abundant and diverse signatures of eukaryotes with <80% sequence similarity to sequences in GenBank. This suggests the presence of significant novel eukaryotic diversity, particularly in hypersaline Hamelin Pool. There was evidence of vertical structuring of protist populations and foraminiferal diversity was highest in bioturbated/clotted thrombolite mats of Highborne Cay.     

Prenylated hydroquinones: contact allergens from trichomes of Phacelia minor and P. parryi

Geranylgeranylhydroquinone and 1-oxofarnesylhydroquinone were identified as contact allergens in trichomes of Phacelia minor and P. parryi.     

Starch grain morphology of the seagrasses Halodule wrightii, Ruppia maritima, Syringodium filiforme, and Thalassia testudinum

Starch grains are a ubiquitous component of plants that have been used in tandem with phytoliths, pollen, and macrofossils to reconstruct past floral diversity. This tool has yet to be fully explored for aquatic plants, specifically seagrasses, which lack phytoliths and are rarely preserved as macrofossils or pollen. If starch grains in seagrasses are morphologically distinct, this method has the potential to improve seagrass identification in the fossil record in such cases where its starch is preserved (e.g. scratches and occlusal surfaces of tooth enamel from seagrass consumers). The goals of this study were twofold: (1) to determine if starch is present in seagrass material and (2) to assess how starch grain morphology differs between different seagrasses.This study focused on four abundant and ecologically distinct seagrasses from the Caribbean: Halodule wrightii, Ruppia maritima, Syringodium filiforme, and Thalassia testudinum. Starch grains were observed in all species except S. filiforme. Grains from H. wrightii are typically observed in side-on orientation, are sub-round to angular, and are fairly small (3-19 μm, end-on). Grains of R. maritima are small spherical grains (4-8 μm) that have a centric hilum and a straight extinction cross with a median angle between the arms of 90°. Grains from T. testudinum are large (9-31 μm, end-on), conical in side-on and round/sub-round in end-on orientation, have a slightly eccentric hilum with an obvious particle, and prominent lamellae.Visual assessment and comparative statistics demonstrate that the morphology of starch grains from T. testudinum, R. maritima, and H. wrightii are significantly different. With more extensive research, there is potential for the positive identification of starch grains from an unknown seagrass. The ability to identify seagrass from starch grains could facilitate the identification of seagrasses in the fossil record and supply information on seagrass evolution and distribution, climate effects on seagrass distribution, and the diets of seagrass consumers.     

Seed dormancy,soil type and protective shelters influence seedling emergence at Shark Bay,Western Australia: Insight into global dryland revegetation

Seedling emergence is a major constraint on dryland revegetation success. In this study, we investigated seedling emergence of six framework shrub species as influenced by seed treatment, soil type and protective shelters using a large field trial in arid Western Australia. We observed the main effects of seed treatment and soil type to account for the majority of the variation in emergence. For species that exhibit pronounced dormancy, we found emergence of dormancy‐alleviated or treated (T) seed to be significantly greater than dormant or untreated (UT) seed, with responses varying across species (e.g. 41 times greater for Acacia ligulata Benth., and 10 times greater for Stylobasium spathulatum Desf.). For shallowly or nondormant species like Senna glutinosa (DC) Randall, UT seed emergence was slightly greater than for T seed. Compared to subsoil, topsoil was more receptive to infiltration (3.44 vs. 0.38 mm/min), and less prone to compaction (1.24 vs. 1.67 g/cm³) and crusting (0.6 vs. 1.3 kg/cm²); however, subsoil had greater moisture retention. Shelters failed to benefit soil moisture retention in either soil type, but enhanced emergence for most species. This study provides insight into how various cost‐effective treatments can be utilized to manipulate seed dormancy to optimize seedling emergence, the intrinsic value of topsoil as a superior growth medium and the benefit of novel, low‐cost shelters for enhancing seedling emergence. In arid environments, sowing T seed in combination with UT seed increases the likelihood of capitalizing on inherently variable precipitation events.     

Glycoprotein 60 diversity in C. hominis and C. parvum causing human cryptosporidiosis in NSW, Australia

Management and control of cryptosporidiosis in human requires knowledge of Cryptosporidium species contributing to human disease. Markers that are able to provide information below the species level have become important tools for source tracking. Using the hypervariable surface antigen, glycoprotein 60 (GP60), C. hominis (n = 37) and C. parvum (n = 32) isolates from cryptosporidiosis cases in New South Wales, Australia, were characterised. Extensive variation was observed within this locus and the isolates could be divided into 8 families and 24 different subtypes. The subtypes identified have global distributions and indicate that anthroponotic and zoonotic transmission routes contribute to sporadic human cryptosporidiosis in NSW.     

Age and Growth of the Whiskery Shark, Furgaleus macki, from Southwestern Australia

Age and growth of the whiskery shark, Furgaleus macki, from southwestern Australia were examined using vertebral ageing and tag-recapture data. The readability of bands on the vertebral centra varied markedly between individuals. Four readers were used to make band counts, with the most experienced reader having the lowest index of average percent error and the highest level of agreement with final counts. Marginal increment analysis indicated that opaque bands form in January. With parturition occurring from August to October, size data suggests that the first band is probably formed 15–17 months after birth. The age at maturity was estimated to be 4.5 years for males, and 6.5 years for females. The oldest male was 10.5 years, and oldest female was 11.5 years. Von Bertalanffy growth parameters for males were L =121.5cm fork length, K=0.423 year^–1, t ₀=–0.472 years, were L =120.7cm fork length, K=0.369 year^–1, t ₀=–0.544 years for females, and were L =118.1cm fork length, K=0.420 year^–1, t ₀=–0.491 years for combined sexes. Data from a tag recapture study were analysed using a maximum likelihood method to verify the estimates of growth parameters from vertebral ageing. Von Bertalanffy growth parameters from the tag recapture study were L =128.2cm fork length, K=0.288 year^–1, t ₀=–0.654 years. The two methods of estimating growth parameters produced similar results, with rapid growth until approximately 5 years of age, after which there was little increase in length.     

Annual spawning of the hydrothermal vent mussel, Bathymodiolus azoricus, under controlled aquarium, conditions at atmospheric pressure

Dense macrofaunal communities of modioliform mussels are a major component of many hydrothermal vent and cold seep ecosystems. The hydrothermal vent mussel Bathymodiolus azoricus, that dominates hydrothermal vent communities near the Azores Triple Junction, can be maintained in aquaria at atmospheric pressure. Cages containing these mussels were placed over diffuse vent outlets and recovered at different times. Cages recovered in January 2003 contained mussels with ripe gonads while those recovered in July, August and November 2001 and in April 2003 did not. Mussels collected post-spawning in April 2003 spawned in the aquaria in January 2004. Young mussels recruited to the cages in April 2003. The data support a main single period of spawning and of juvenile recruitment for B. azoricus.     

The effect of Stratiotes aloides L. and nutrients on the growth rate of Lemna minor L.

We tried to find the reasons for frequently observed low biomass of Lemna minor among floating rosettes of Stratiotes aloides as opposed to the abundance of Spirodela polyrhiza in these stands. The effect of S. aloides on L. minor was analysed in outdoor culture experiments. The duckweed collected from among dense stands of the floating form of S. aloides (test sites) showed a significantly lower growth rate than that from stands free of the water soldier (control sites). Water from the latter stands was significantly richer in nitrate-nitrogen and ammonium-nitrogen, but not in soluble reactive phosphorus suggesting possible nutrient depletion as a reason for observed differences. Growth rates of L. minor were indeed significantly correlated with in situ concentrations of all dissolved nutrients and showed saturation at around 0.18 mg N-NH₄ dm⁻³ and 0.05 mg SRP dm⁻³ above which the growth rates did not increase significantly. We used stepwise regression to test the combined effect of nutrients and the presence/absence of S. aloides on the growth rate of L. minor. The regression showed a negligible effect of ammonium ions and nitrates on the growth rate of the duckweed. At low concentrations of soluble reactive phosphorus the presence of both the S. aloides and SRP concentrations significantly affected the growth of L. minor. Above saturating SRP concentrations none of the two effects was significant. Results of our experiments seem to suggest that the presence of S. aloides exerts a negative effect on the growth of L. minor only at low concentrations of soluble phosphorus. Since the depletion of nutrients in water by the floating form of S. aloides is unlikely, we point to S. polyrhiza as a superior competitor for nutrients and suggest a possible allelopathic effect of the water soldier on L. minor.     

Isolation and diversity of planctomycetes from the sponge Niphates sp., seawater, and sediment of Moreton Bay, Australia

Planctomycetes are ubiquitous in marine environment and were reported to occur in association with multicellular eukaryotic organisms such as marine macroalgae and invertebrates. Here, we investigate planctomycetes associated with the marine sponge Niphates sp. from the sub-tropical Australian coast by assessing their diversity using culture-dependent and -independent approaches based on the 16S rRNA gene. The culture-dependent approach resulted in the isolation of a large collection of diverse planctomycetes including some novel lineages of Planctomycetes from the sponge as well as sediment and seawater of Moreton Bay where this sponge occurs. The characterization of these novel planctomycetes revealed that cells of one unique strain do not possess condensed nucleoids, a phenotype distinct from other planctomycetes. In addition, a culture-independent clone library approach identified unique planctomycete 16S rRNA gene sequences closely related to other sponge-derived sequences. The analysis of tissue of the sponge Niphates sp. showed that the mesohyl of the sponge is almost devoid of microbial cells, indicating this species is in the group of ‘low microbial abundant’ (LMA) sponges. The unique planctomycete 16S rRNA gene sequences identified in this study were phylogenetically closely related to sequences from LMA sponges in other published studies. This study has revealed new insights into the diversity of planctomycetes in the marine environment and the association of planctomycetes with marine sponges.     

The Biology of Tiger Sharks, Galeocerdo Cuvier, in Shark Bay, Western Australia: Sex Ratio, Size Distribution, Diet, and Seasonal Changes in Catch Rates

Tiger sharks, Galeocerdo cuvier, are apex predators in a variety of nearshore ecosystems throughout the world. This study investigates the biology of tiger sharks in the shallow seagrass ecosystem of Shark Bay, Western Australia. Tiger sharks (n = 252) were the most commonly caught species (94%) compared to other large sharks. Tiger sharks ranged from 148–407cm TL. The overall sex ratio was biased towards females (1.8:1), but the sex ratio of mature animals (> 300cm TL) did not differ from 1:1. Contrary to previous accounts, tiger sharks were caught more often in all habitats during daylight hours than at night. Tiger shark catch rates were highly correlated with water temperature and were highest when water temperatures were above 19°C. The seasonal abundance of tiger sharks is correlated to both water temperature and the occurrence of their main prey: sea snakes and dugongs, Dugong dugon. Stomach contents analysis indicated that sea turtles and smaller elasmobranchs were also common prey. The importance of major seagrass grazers (dugongs and green sea turtles, Chelonia mydas) in the diet of tiger sharks suggests the possibility that these sharks are keystone predators in this ecosystem.