Diversity and Activity of Diazotrophs in Great Barrier Reef Surface Waters.

Discrepancies between bioavailable nitrogen (N) concentrations and phytoplankton growth rates in the oligotrophic waters of the Great Barrier Reef (GBR) suggest that undetermined N sources must play a significant role in supporting primary productivity. One such source could be biological dinitrogen (N2) fixation through the activity of "diazotrophic" bacterioplankton. Here, we investigated N2 fixation and diazotroph community composition over 10° S of latitude within GBR surface waters. Qualitative N2 fixation rates were found to be variable across the GBR but were relatively high in coastal, inner and outer GBR waters, reaching 68 nmol L-1 d-1. Diazotroph assemblages, identified by amplicon sequencing of the nifH gene, were dominated by the cyanobacterium Trichodesmium erythraeum, γ-proteobacteria from the Gamma A clade, and δ-proteobacterial phylotypes related to sulfate-reducing genera. However, diazotroph communities exhibited significant spatial heterogeneity, correlated with shifts in dissolved inorganic nutrient concentrations. Specifically, heterotrophic diazotrophs generally increased in relative abundance with increasing concentrations of phosphate and N, while Trichodesmium was proportionally more abundant when concentrations of these nutrients were low. This study provides the first in-depth characterization of diazotroph community composition and N2 fixation dynamics within the oligotrophic, N-limited surface waters of the GBR. Our observations highlight the need to re-evaluate N cycling dynamics within oligotrophic coral reef systems, to include diverse N2 fixing assemblages as a potentially significant source of dissolved N within the water column.

The cycling and fate of terrestrially-derived sediments and nutrients in the coastal zone of the Great Barrier Reef shelf.

The coastal zone of the Great Barrier Reef shelf, with an area of 30,000 km(2) and a water volume of 300 km(3), receives an average annual input of sediment on the order of 14-28 Mty(-1)--an estimated two- to fourfold increase since European settlement. There is considerable concern about the impact and ultimate fate of terrestrially-derived nutrients entering the Great Barrier Reef World Heritage Area (GBRWHA). Analysis of current data suggests that microbial communities in coastal waters and in unconsolidated sediments metabolise nutrients equivalent to the entire dissolved and particulate nutrient load debouched from land. River-derived nutrients account for 40-80% of the carbon, 13-30% of the nitrogen, and 2-5% of the phosphorus necessary to support the observed rates of benthic and pelagic mineralisation in Princess Charlotte Bay in the far north, and in Rockingham Bay and Missionary Bay in the central section, of the GBRWHA. Nearly all nitrogen is ultimately returned to the atmosphere via denitrification. There is little net burial of nutrients in subtidal sediments. These budget estimates are based on a sparse data set, but it is clear that marine sources of nutrients (N-fixation by pelagic and benthic cyanobacteria) must be important, but the magnitude of these sources is poorly known and likely to be highly variable in space and time. Data from sediment trap deployments suggest that, despite significant re-suspension, sedimentation fluxes are sufficient to balance benthic mineralisation rates. Most organic material deposited to the benthos appears to be flocculent or colloidal aggregates, perhaps formed via microbial mediation and exudation of extra-cellular material. The geophysical dynamics of the coastal boundary layer plays an important role in concentrating biological and biogeochemical activity within a shallow, narrow coastal zone. Mangroves and tidal flats are small in area, but trap, transform, and store a disproportionate amount of sediment and organic matter within the GBRWHA. The highly efficient use of terrestrially-derived nutrients by benthic and pelagic microbes in the coastal zone helps to explain why coral reefs on the middle and outer shelf have remained relatively unscathed despite a significant increase in sediment delivery.

Outbreak of coral-eating Crown-of-Thorns creates continuous cloud of larvae over 320 km of the Great Barrier Reef.

Coral reefs are in decline worldwide due to a combination of local and global causes. Over 40% of the recent coral loss on Australia's Great Barrier Reef (GBR) has been attributed to outbreaks of the coral-eating Crown-of-Thorns Seastar (CoTS). Testing of the hypotheses explaining these outbreaks is hampered by an inability to investigate the spatio-temporal distribution of larvae because they resemble other planktotrophic echinoderm larvae. We developed a genetic marker and tested it on 48 plankton samples collected during the 2014 spawning season in the northern GBR, and verified the method by PCR amplification of single larva. Surprisingly, most samples collected contained CoTS larvae. Larvae were detected 100 km south of current outbreaks of adult seastars, highlighting the potential for rapid expansion of the outbreak. A minimum estimate suggested that larvae numbers in the outbreak area (>10(10)) are about 4 orders of magnitude higher than adults (~10(6)) in the same area, implying that attempts to halt outbreaks by removing adults may be futile.

A synthesis of dominant ecological processes in intensive shrimp ponds and adjacent coastal environments in NE Australia.

One of the key environmental concerns about shrimp farming is the discharge of waters with high levels of nutrients and suspended solids into adjacent waterways. In this paper we synthesize the results of our multidisciplinary research linking ecological processes in intensive shrimp ponds with their downstream impacts in tidal, mangrove-lined creeks. The incorporation of process measurements and bioindicators, in addition to water quality measurements, improved our understanding of the effect of shrimp farm discharges on the ecological health of the receiving water bodies. Changes in water quality parameters were an oversimplification of the ecological effects of water discharges, and use of key measures including primary production rates, phytoplankton responses to nutrients, community shifts in zooplankton and delta15N ratios in marine plants have the potential to provide more integrated and robust measures. Ultimately, reduction in nutrient discharges is most likely to ensure the future sustainability of the industry.

Effect of short-term macrophage depletion in the development of posterior capsule opacification in rodents.

AIM: To evaluate the role of macrophages in the development of posterior capsule opacification (PCO). METHODS: For this purpose, an extracapsular lens extraction was performed in 18 consecutive Sprague-Dawley rats. Animals were treated with liposomal clodronate (Cl(2)MDP-lip-treated group, n = 10) or phosphate-buffered saline (PBS) (control group, n = 8) 1 day preoperatively and on the first day postoperatively, and sacrificed 3 days postoperatively. Masked clinical, light microscopy and immunohistochemistry studies were conducted. The Fisher exact test and randomisation test were used to assess statistically differences between groups. RESULTS: A statistically significant reduction in the number of macrophages (ED1+, ED7+, ED8+) was found in the Cl(2)MDP-lip-treated group compared with the PBS-lip-treated group (p = 0.048, p = 0.004, p = 0.027, respectively). There were no statistically significant differences with regards to the presence/absence of central opacification (p = 0.29) and capsular wrinkling (p = 0.21) as detected clinically between groups. Similarly, a qualitative evaluation of the degree of PCO with regards to lens epithelial cell (LEC) proliferation, capsular wrinkling and Soemmerring ring formation showed no statistically significance between groups (p = 0.27, p = 0.061, p = 1.0, respectively). However, a statistically significant reduction in the number of lens epithelial cells (LEC) counted in the centre of the posterior capsule was found in the Cl(2)MDP-lip-treated group (p = 0.009). CONCLUSION: Depletion of macrophages was accompanied by a reduction in LEC in the centre of the posterior capsule in rodents.

Histological and ultrastructural studies of the marsupial Didelphis albiventris Peyer's patches.

Differing from the studied Eutheria the white belly opossum Peyer's patches do not present a conspicuous dome. M cells are located in the inner layer of bilaminal formed at the bottom of the villi. A great variation in the morphology of M cells was observed. The enterocytes located at the epithelial inner layer may present endocytic vesicles, and the microvilli are shorter than the microvilli of enterocytes lining the small intestine. As these morphological aspects have been described to exist in the enterocytes of the lactent opossum small intestine it was surmised that the opossum Peyer's patches special epithelium could represent the persistence in adult animals of a cellular pattern established before the intestinal maturation had occurred.

Lung capillary albumin leak in oxygen toxicity. A quantitative immunocytochemical study.

The study is based on the hypothesis that lung endothelial cell permeability increases in 100% oxygen and predates the appearance of microscopically visible interstitial edema. Rats were exposed to either 100% oxygen or air in a chamber. Endogenous albumin was used as an index of permeability and measured by electron microscopic colloidal gold linked immunocytochemistry, quantified by systematic random methods. Albumin staining was expressed as relative albumin concentration (RAC), the ratio of gold particles (x 100) per point counted (gp.10(2)/pt) relating to each component. The RAC in lung perivascular/peribronchial interstitial ground substance after 24 h of hyperoxia was five times more than that of rats exposed to air for the same interval. The median value (interquartile ranges) for the oxygen-exposed group was 92.4 (39.5, 149.6) gp. 10(2)/pt compared with 14.7 (6.6, 25.9) gp. 10(2)/pt for the air-exposed group. After 60 h of 100% oxygen, the RAC was 103.4 (65.5, 148.9) gp. 10(2)/pt (60-h air exposed RAC was 11.6 (8.7, 60.4) gp. 10(2)/pt), no different from 24-h exposures. These results suggest that there was a significant leak of albumin to the perivascular/peribronchial interstitium by 24 h of exposure to 100% oxygen, which would indicate endothelial cell permeability to albumin increases earlier than has previously been reported.

Identification of a novel phosphonocarboxylate inhibitor of Rab geranylgeranyl transferase that specifically prevents Rab prenylation in osteoclasts and macrophages.

Nitrogen-containing bisphosphonate drugs inhibit bone resorption by inhibiting FPP synthase and thereby preventing the synthesis of isoprenoid lipids required for protein prenylation in bone-resorbing osteoclasts. NE10790 is a phosphonocarboxylate analogue of the potent bisphosphonate risedronate and is a weak anti-resorptive agent. Although NE10790 was a poor inhibitor of FPP synthase, it did inhibit prenylation in J774 macrophages and osteoclasts, but only of proteins of molecular mass approximately 22-26 kDa, the prenylation of which was not affected by peptidomimetic inhibitors of either farnesyl transferase (FTI-277) or geranylgeranyl transferase I (GGTI-298). These 22-26-kDa proteins were shown to be geranylgeranylated by labelling J774 cells with [(3)H]geranylgeraniol. Furthermore, NE10790 inhibited incorporation of [(14)C]mevalonic acid into Rab6, but not into H-Ras or Rap1, proteins that are modified by FTase and GGTase I, respectively. These data demonstrate that NE10790 selectively prevents Rab prenylation in intact cells. In accord, NE10790 inhibited the activity of recombinant Rab GGTase in vitro, but did not affect the activity of recombinant FTase or GGTase I. NE10790 therefore appears to be the first specific inhibitor of Rab GGTase to be identified. In contrast to risedronate, NE10790 inhibited bone resorption in vitro without markedly affecting osteoclast number or the F-actin "ring" structure in polarized osteoclasts. However, NE10790 did alter osteoclast morphology, causing the formation of large intracellular vacuoles and protrusion of the basolateral membrane into large, "domed" structures that lacked microvilli. The anti-resorptive activity of NE10790 is thus likely due to disruption of Rab-dependent intracellular membrane trafficking in osteoclasts.