Gambierdiscus (Dinophyceae) species diversity in the Flower Garden Banks National Marine Sanctuary,Northern Gulf of Mexico,USA

Globally, ciguatera fish poisoning (CFP) is the principal cause of non-bacterial illness associated with seafood consumption. The toxins (ciguatoxins) responsible for CFP are produced by dinoflagellates in the genus Gambierdiscus, which are endemic to tropical and sub-tropical areas. Ciguatoxins are lipophilic and bioaccumulate in marine food webs, typically reaching their highest concentrations in fish. Following a CFP event in 2008, the U.S. Food and Drug Administration (USFDA) issued a ciguatera toxin alert that included fish harvested in the northern Gulf of Mexico in and near the Flower Garden Banks National Marine Sanctuary (FGBNMS). The East Flower Garden Bank (EFGB) and West Flower Garden Bank (WFGB) are characterized by thriving coral communities that support Gambierdiscus growth. This study was undertaken specifically to document the diversity of Gambierdiscus species present in the sanctuary that may be sources of ciguatoxins entering the food web. Samples collected from the FGBNMS over a three year period were screened using species-specific polymerase chain reaction assays. A diverse assemblage of Gambierdiscus species was distributed to depths of >45 m, a new depth record for Gambierdiscus. Gambierdiscus belizeanus, Gambierdiscus caribaeus, Gambierdiscus carolinianus, Gambierdiscus carpenteri and Gambierdiscus ribotype 2 were all found on both East and West FGB with Gambierdiscus ruetzleri also recorded from the WFGB. The most common species was G. carolinianus, originally identified from samples collected between 35 and 40 m off the coast of NC, USA. Our findings are consistent with recent physiological studies showing that some Gambierdiscus species can grow year round at the temperatures and salinities at the FGBNMS and at light levels as low as 10 μmol photons m⁻² s⁻¹. Such irradiances are estimated to occur in the FGBNMS at depths of ∼70–80 m. The consistent recovery of Gambierdiscus species from deep sampling sites in areas known to produce ciguatoxic fish signals a substantial change in our concept of suitable habitats for Gambierdiscus to include depths greater than 50 m.     

Fukuyoa paulensis gen. et sp. nov., a New Genus for the Globular Species of the Dinoflagellate Gambierdiscus (Dinophyceae)

The marine epiphytic dinoflagellate Gambierdiscus is a toxicologically important genus responsible for ciguatera fish poisoning, the principal cause of non-bacterial illness associated with fish consumption. The genus currently contains species exhibiting either globular or anterior-posteriorly compressed morphologies with marked differences in cell shape and plate arrangement. Here we report a third globular, epiphytic and tychoplanktonic species from the coasts of Ubatuba, Brazil. The new species can be distinguished from G. yasumotoi and G. ruetzleri by its broader first apical plate that occupies a larger portion of the epitheca. Accordingly, phylogenetic trees from small subunit (SSU) and large subunit (LSU) ribosomal DNA sequences also showed strongly supported separation of the new species from the G. yasumotoi / G. ruetzleri group albeit with short distance. The molecular phylogenies, which included new sequences of the planktonic species Goniodoma polyedricum, further indicated that the globular species of Gambierdiscus formed a tight clade, clearly separated (with strong bootstrap support) from the clade of lenticular species including the type for Gambierdiscus. The morphological and molecular data in concert support the split of Gambierdiscus sensu lato into two genera. Gambierdiscus sensu stricto should be reserved for the species with lenticular shapes, highly compressed anterioposteriorly, with short-shank fishhook apical pore plate, large 2'' plate, low and ascending cingular displacement, and pouch-like sulcal morphology. The new genus name Fukuyoa gen. nov. should be applied to the globular species, slightly laterally compressed, with long-shank fishhook apical pore plate, large 1'' plate, greater and descending cingular displacement, and not pouch-like vertically-oriented sulcal morphology. Fukuyoa contains the new species Fukuyoa paulensis gen. et sp. nov., and F. yasumotoi comb. nov. and F. ruetzleri comb. nov.     

Growth of eight Gambierdiscus (Dinophyceae) species: Effects of temperature,salinity and irradiance

Little is known about how the growth of individual Gambierdiscus species responds to environmental factors. This study examined the effects of temperature (15–34 °C), salinity (15–41) and irradiance (2–664 μmol photons m⁻² s⁻¹) on growth of Gambierdiscus: G. australes, G. belizeanus, G. caribaeus, G. carolinianus, G. carpenteri, G. pacificus and G. ruetzleri and one putative new species, Gambierdiscus ribotype 2. Depending on species, temperatures where maximum growth occurred varied between 26.5 and 31.1 °C. The upper and lower thermal limits for all species were between 31–34 °C and 15–21 °C, respectively. The shapes of the temperature vs. growth curves indicated that even small differences of 1–2 °C notably affected growth potentials. Salinities where maximum growth occurred varied between 24.7 and 35, while the lowest salinities supporting growth ranged from <14 to 20.9. These data indicated that Gambierdiscus species are more tolerant of lower salinities than is generally appreciated. Growth of all species began to decline markedly as salinities exceed 35.1–39.4. The highest salinity tested in this study (41), however, was lethal to only one species, Gambierdiscus ribotype 2. The combined salinity data indicated that differences in salinity regimes may affect relative species abundances and distributions, particularly when salinities are <20 and >35. All eight Gambierdiscus species were adapted to relatively low light conditions, exhibiting growth maxima at 50–230 μmol photons m⁻² s⁻¹ and requiring only 6–17 μmol photons m⁻² s⁻¹ to maintain growth. These low light requirements indicate that Gambierdiscus growth can occur up to 150 m depth in tropical waters, with optimal light regimes often extending to 75 m. The combined temperature, salinity and light requirements of Gambierdiscus can be used to define latitudinal ranges and species-specific habitats, as well as to inform predictive models.     

Effects of temperature,salinity and composition of the dinoflagellate assemblage on the growth of Gambierdiscus carpenteri isolated from the Great Barrier Reef

Increases in reported incidence of ciguatera fish poisoning (hereafter ciguatera) have been linked to warmer sea temperatures that are known to trigger coral bleaching events. The drivers that trigger blooms of ciguatera-causing dinoflagellates on the Great Barrier Reef (GBR) are poorly understood. This study investigated the effects of increased temperatures and lowered salinities, often associated with environmental disturbance events, on the population growth of two strains of the potentially ciguatera-causing dinoflagellate, Gambierdiscus carpenteri (NQAIF116 and NQAIF380). Both strains were isolated from the central GBR with NQAIF116 being an inshore strain and NQAIF380 an isolate from a stable environment of a large coral reef aquarium exhibit in ReefHQ, Townsville, Australia. Species of Gambierdiscus are often found as part of a mixed assemblage of benthic toxic dinoflagellates on macroalgal substrates. The effect of assemblage structure of dinoflagellates on the growth of Gambierdiscus populations has, however, not been explored. The study, therefore investigated the growth of G. carpenteri within mixed assemblages of benthic dinoflagellates. Population growth was monitored over a period of 28 days under three salinities (16, 26 and 36) and three temperature (24, 28 and 34 °C) conditions in a fully crossed experimental design. Temperature and salinity had a significant effect on population growth. Strain NQAIF380 exhibited significantly higher growth at 28 °C compared to strain NQAIF116, which had highest growth at 24 °C. When strain NQAIF116 was co-cultured with the benthic dinoflagellates, Prorocentrum lima and Ostreopsis sp., inhibitory effects on population growth were observed at a salinity of 36. In contrast, growth stimulation of G. carpenteri (strain NQAIF116) was observed at a salinity of 26 and particularly at 16 when co-cultured with Ostreopsis-dominated assemblages. Range expansion of ciguatera-causing dinoflagellates could lead to higher frequency of reported ciguatera illness in populated temperate Australian regions, outside the tropical range of the GBR. Therefore, the findings on salinity and temperature tolerance of two strains of G. carpenteri indicates potential adaptability to different local environmental conditions. These are baseline data for future investigations into the potential southward range expansion of ciguatera-causing dinoflagellates originating from the GBR.     

Production of ciguatoxin and maitotoxin by strains of Gambierdiscus australes,G. pacificus and G. polynesiensis (Dinophyceae) isolated from Rarotonga,Cook Islands

The isolation and culturing of toxic Gambierdiscus species is desirable for obtaining reliable sources of toxins responsible for Ciguatera Fish Poisoning (CFP) in order to provide reference materials and purified standards for chemical analyses and bioassays. Gambierdiscus polynesiensis CAWD212, G. pacificus CAWD213 and G. australes CAWD149 and 216 have been isolated from samples of sea water from the Cook Islands. G. polynesiensis produced ciguatoxins (CTX; 18.2 pg per cell) and G. australes CAWD149 produced maitotoxin (MTX; 8.3 pg per cell) and in both extracts putative MTX-3 was detected. G. pacificus also produced putative MTX-3. The toxicities of extracts of G. polynesiensis and G. pacificus were similar by i.p. injection (LD_50s of 1.0 mg/kg and 0.8 mg/kg respectively), but when administered by gavage, the MTX-3 analogue producing G. pacificus (LD₅₀ 251 mg/kg) was 30-times less toxic than the CTX-producing G. polynesiensis (LD₅₀ 7.9 mg/kg). Consumption of fish contaminated by CFP has caused illness and even death in the Cook Islands and throughout the Pacific region and is now considered a globally neglected disease. Large-scale cultures of these organisms are now available, and isolation of the toxins they produce is in progress.     

The prevalence of benthic dinoflagellates associated with ciguatera fish poisoning in the central Red Sea

This study confirms the presence of the toxigenic benthic dinoflagellates Gambierdiscus belizeanus and Ostreopsis spp. in the central Red Sea. To our knowledge, this is also the first report of these taxa in coastal waters of Saudi Arabia, indicating the potential occurrence of ciguatera fish poisoning (CFP) in that region. During field investigations carried out in 2012 and 2013, a total of 100 Turbinaria and Halimeda macroalgae samples were collected from coral reefs off the Saudi Arabian coast and examined for the presence of Gambierdiscus and Ostreopsis, two toxigenic dinoflagellate genera commonly observed in coral reef communities around the world. Both Gambierdiscus and Ostreopsis spp. were observed at low densities (<200 cells g⁻¹ wet weight algae). Cell densities of Ostreopsis spp. were significantly higher than Gambierdiscus spp. at most of the sampling sites, and abundances of both genera were negatively correlated with seawater salinity. To assess the potential for ciguatoxicity in this region, several Gambierdiscus isolates were established in culture and examined for species identity and toxicity. All isolates were morphologically and molecularly identified as Gambierdiscus belizeanus. Toxicity analysis of two isolates using the mouse neuroblastoma cell-based assay for ciguatoxins (CTX) confirmed G. belizeanus as a CTX producer, with a maximum toxin content of 6.50 ± 1.14 × 10⁻⁵ pg P-CTX-1 eq. cell⁻¹. Compared to Gambierdiscus isolates from other locations, these were low toxicity strains. The low Gambierdiscus densities observed along with their comparatively low toxin contents may explain why CFP is unidentified and unreported in this region. Nevertheless, the presence of these potentially toxigenic dinoflagellate species at multiple sites in the central Red Sea warrants future study on their possible effects on marine food webs and human health in this region.     

The effects of light intensity on the growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a form of seafood poisoning that is widespread in tropical, subtropical and temperate regions worldwide. The distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3 have been reported for the waters surrounding the main island of Japan. To explore the bloom dynamics and the vertical distribution of these Japanese species and phylotypes of Gambierdiscus, the effects of light intensity on their growth were tested, using a photoirradiation-culture system. The relationship between the observed growth rates and light intensity conditions for the four species/phylotypes were formulated at R > 0.92 (p < 0.01) using regression analysis and photosynthesis-light intensity (P-L) model. Based on this equation, the optimum light intensity (L_max) and the semi-optimum light intensity range (L_s-opt) that resulted in the maximum growth rate (μ_max) and ≥80% μ _max values of the four species/phylotypes, respectively, were as follows: (1) the L_max and L_s-opt of G. australes were 208 μmol photons m⁻² s⁻¹ and 91–422 μmol photons m⁻² s⁻¹, respectively; (2) those of G. scabrosus were 252 and 120–421 μmol photons m⁻² s⁻¹, respectively; (3) those of Gambierdiscus sp. type 2 were 192 and 75–430 μmol photons m⁻² s⁻¹, respectively; and (4) those of Gambierdiscus sp. type 3 were ≥427 and 73–427 μmol photons m⁻² s⁻¹, respectively. All four Gambierdiscus species/phylotypes required approximately 10 μmol photons m⁻² s⁻¹ to maintain growth. The light intensities in coastal waters at a site in Tosa Bay were measured vertically at 1 m intervals once per season. The relationships between the observed light intensity and depth were formulated using Beer’s Law. Based on these equations, the range of the attenuation coefficients at Tosa Bay site was determined to be 0.058–0.119 m⁻¹. The values 1700 μmol photons m⁻² s⁻¹, 500 μmol photons m⁻² s⁻¹, and 200 μmol photons m⁻² s⁻¹ were substituted into the equations to estimate the vertical profiles of light intensity at sunny midday, cloudy midday and rainy midday, respectively. Based on the regression equations coupled with the empirically determined attenuation coefficients for each of the four seasons, the ranges of the projected depths of L_max and L_s-opt for the four Gambierdiscus species/phylotypes under sunny midday conditions, cloudy midday conditions, and rainy midday conditions were 12–38 m and 12–54 m, 1–16 m and 1–33 m, and 0 m and 0–16 m, respectively. These results suggest that light intensity plays an important role in the bloom dynamics and vertical distribution of Gambierdiscus species/phylotypes in Japanese coastal waters.     

A red tide off the Myanmar coast: Morphological and genetic identification of the dinoflagellate composition

A red tide composed mainly of two Prorocentrum species was found near Kadan Island, southern Myanmar coast, on March 14, 2012. This is the first record of a red tide in the Myanmar coastal area. The red tide included three different harmful dinoflagellates: Prorocentrum rhathymum, Prorocentrum shikokuense and Alexandrium affine. Strains of these species were successfully cultured and examined with detailed morphological observation and DNA (28S rRNA gene) analyses. The strains were subjected to growth experiments under different temperatures to understand their growth capabilities. The experiments were carried out at four different temperature regimes (15, 20, 25 and 30 °C). A. affine exhibited low tolerance for the low temperature regime (15 °C), despite records of its presence in northern temperate regions, indicating this strain is adapted to the tropical environment in Myanmar. P. rhathymum and P. shikokuense exhibited broad tolerance to all given temperature ranges and showed high division rates, providing the physiological basis to form red tides.     

Cyclestheria hislopi (Crustacea: Branchiopoda): A group of morphologically cryptic species with origins in the Cretaceous

Cyclestheria hislopi is thought to be the only extant species of Cyclestherida. It is the sister taxon of all Cladocera and displays morphological characteristics intermediate of Spinicaudata and Cladocera. Using one mitochondrial (COI) and two nuclear (EF1α and 28S rRNA) markers, we tested the hypothesis that C. hislopi represents a single circumtropic species. South American (French Guiana), Asian (India, Indonesia, Singapore) and several Australian populations were included in our investigation. Phylogenetic and genetic distance analyses revealed remarkable intercontinental genetic differentiation (uncorrected p-distances COI > 13%, EF1α > 3% and 28S > 4%). Each continent was found to have at least one distinct Cyclestheria species, with Australia boasting four distinct main lineages which may be attributed to two to three species. The divergence of these species (constituting crown group Cyclestherida) was, on the basis of phylogenetic analyses of COI and EF1α combined with molecular clock estimates using several fossil branchiopod calibration points or a COI substitution rate of 1.4% per million years, dated to the Cretaceous. This was when the South American lineage split from the Asian–Australian lineage, with the latter diverging further in the Paleogene. Today’s circumtropic distribution of Cyclestheria may be best explained by a combination of Gondwana vicariance and later dispersal across Asia and Australia when the tectonic plates of the two continents drew closer in the early Miocene. The lack of morphological differentiation that has taken place in this taxon over such a long evolutionary period contrasts with the high level of differentiation and diversification observed in its sister taxon the Cladocera. Further insights into the evolution of Cyclestheria may help us to understand the evolutionary success of the Cladocera.     

Effects of temperature,salinity and their interaction on growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a seafood poisoning that is widespread in tropical, subtropical and temperate regions of the world. In the main island of Japan, distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3, have been reported. To discuss the bloom dynamics of these Japanese species/phylotypes of Gambierdiscus, first we tested six culture media to optimize growth conditions and then clarified the effects of temperature and salinity and temperature–salinity interactions on growth. All strains of the species/phylotypes tested showed the highest cell yields when they were cultivated in IMK/2 medium. G. australes, G. scabrosus and Gambierdiscus sp. type 2 grew in the range 17.5–30 °C, whereas Gambierdiscus sp. type 3 grew in 15–25 °C. The semi-optimal temperature ranges (≥80% of the maximal growth rate) of the former three species/phylotypes were 19–28 °C, 24–31 °C and 21–28 °C, respectively, whereas that of the latter phylotype was 22–25 °C. Hence, Gambierdiscus sp. type 3 may be adapted to relatively lower water temperatures of ≤25 °C. In contrast, G. australes, G. scabrosus and Gambierdiscus sp. type 2 presumably possess adaptability to relatively high water temperatures. The optimal temperature for G. scabrosus was 30 °C, whereas the optimal temperature for the others was 25 °C. G. australes and Gambierdiscus sp. type 3 grew in a salinity range of 25–40 whereas G. scabrosus and Gambierdiscus sp. type 2 grew in salinity 20–40. Furthermore, the semi-optimal salinity range of G. australes, G. scabrosus, Gambierdiscus spp. type 2 and type 3 were salinity 27–38, 24–36, 22–36 and 29–37, respectively. Among the species/phylotypes, G. scabrosus and Gambierdiscus sp. type 2 grew even at salinity 20 where the others did not grow, thus possessing adaptability to low salinity waters. Our results clearly demonstrate that the optimal and tolerable temperature–salinity conditions differ among Japanese Gambierdiscus species/phylotypes. Considering these results, temperature–salinity interactions may play an important role in bloom dynamics and the distribution of the Gambierdiscus species/phylotypes in Japanese coastal waters.     

Recurrent vernal presence of the toxic Alexandrium tamarense/Alexandrium fundyense (Dinoflagellata) species complex in Narragansett Bay,USA

The vernal occurrence of toxic dinoflagellates in the Alexandrium tamarense/Alexandrium fundyense species complex in an enclosed embayment of Narragansett Bay (Wickford Cove, Rhode Island) was documented during 2005 and 2009–2012. This is the first report of regular appearance of the Alexandrium fundyense/Alexandrium tamarense species complex in Narragansett Bay. Thecal plate analysis of clonal isolates using SEM revealed cells morphologically consistent with both Alexandrium tamarense Lebour (Balech) and Alexandrium fundyense Balech. Additionally, molecular analyses confirmed that the partial sequences for 18S through the D1–D2 region of 28S were consistent with the identity of the two Alexandrium species. Toxin analyses revealed the presence of a suite of toxins (C1/2, B1 (GTX-5), STX, GTX-2/3. Neo, and GTX-1/4) in both Alexandrium tamarense (6.31 fmol cell⁻¹ STX equiv.) and Alexandrium fundyense (9.56 fmol cell⁻¹ STX equiv.) isolated from Wickford Cove; the toxicity of a Narragansett Bay Alexandrium peruvianum isolate (1.79 fmol cell⁻¹ STX equiv.) was also determined. Combined Alexandrium tamarense/Alexandrium fundyense abundance in Wickford Cove reached a peak abundance of 1280 cells L⁻¹ (May of 2010), with the combined abundance routinely exceeding levels leading to shellfishing closures in other systems. The toxic Alexandrium tamarense/Alexandrium fundyense species complex appears to be a regular component of the lower Narragansett Bay phytoplankton community, either newly emergent or previously overlooked by extant monitoring programs.     

Different types of fruit damages of three internal apple feeders diagnosed with mitochondrial molecular markers

Three tortricid pests, Grapholita dimorpha (Komai), G. molesta (Busck), and Carposina sasakii (Matsumura), are known as internal apple feeders in Korea. To identify young larvae, this study developed two types of molecular markers from their mitochondrial DNA (mtDNA) sequences. To this end, six different loci of mtDNA were sequenced in G. dimorpha: cytochrome oxidase subunit I (460 bp), cytochrome oxidase subunit II (446 bp), cytochrome b (308 bp), NADH dehydrogenase 3 (585 bp), NADH dehydrogenase 4 (ND4, 835 bp), and 16S rRNA (1300 bp). These sequences were compared with those of G. molesta and C. sasakii in order to develop PCR–RFLP and diagnostic primers. ND4 locus was selected to be used for developing a PCR–RFLP marker. ND4-Swa I digests showed two bands for G. dimorpha, one band for G. molesta, and three bands for C. sasakii. On the other hand, species-specific diagnostic PCR primers were developed using ND4 locus. These markers were then applied to diagnose larvae infesting apples to determine species-specific fruit damage patterns, in which G. dimorpha, G. molesta, and C. sasakii showed different feeding behaviors in terms of their main feeding sites in apple fruits.     

Diversity and spatial clustering of shade trees affect cacao yield and pathogen pressure in Costa Rican agroforests

The importance of the spatial organisation of individuals in explaining species coexistence within a community is widely recognised. However, few analyses of spatial structure have been performed on tropical agroforests.The main objective of this study was to highlight the links between spatial organisation of shade trees on the one hand, and shade tree species richness and cacao yield on the other, using data from 29 cacao agroforests in Costa Rica.A method of spatial statistics, Ripley's K-function, was used to analyse the spatial organisation of shade and cacao trees in the study plots. For each stand, the X and Y coordinates of ≥2.5-m-tall trees were recorded. In each plot we also assessed shade tree species richness and cacao yield (with total number of pods = number of pods damaged by frosty pod rot + number of healthy pods).Three types of stands were identified: the first was characterised by significant clustering of shade trees, the highest shade tree species richness (S = 6), and the highest number of damaged pods (139 pods ha^?1 year^?1). The second type was characterised by random spatial organisation of shade trees. The third type showed a trend towards regular organisation. Species richness of shade trees did not differ significantly between the last two types (S = 4 for both), nor did the number of damaged pods (56 pods ha^?1 year^?1 and 67 pods ha^?1 year^?1 respectively).Although the trends were not statistically significant for all the variables in our data set, the clustered spatial structure appears to favour a synergy between environmental (tree species richness), and provisioning (cacao production) services.     

Epiphytic dinoflagellates in sub-tropical New Zealand,in particular the genus Coolia Meunier

Macroalgae growing in New Zealand's sub-tropical waters were sampled for epiphytic microalgae, in particular dinoflagellates. Four new Coolia isolates collected from sites throughout Northland in January/February 2013 were all identified as C. malayensis as determined by large subunit ribosomal DNA (LSU rDNA) sequence analysis and scanning electron microscopy. C. malayensis, a common dinoflagellate species in New Zealand, was previously reported as C. monotis and isolates held in the Cawthron Institute Culture Collection of Microalgae have now been reclassified based on the DNA sequence data. Toxicity studies of the New Zealand C. malayensis isolates (as determined by intraperitoneal (i.p.) injection of mice) resulted in an LD₅₀ of 80.0 mg/kg for isolate NLD 12 (95% CI: 63.8–101.0 mg/kg) compared to a Malaysian isolate which exhibited low toxicity, with transient effects at 900 mg/kg, but which resulted in the enlargement of the test animal's spleen. Ostreopsis siamensis, a common epiphytic bloom former in northern New Zealand, was found in association with Coolia at sites across Northland. Densest cell concentrations were collected from macroalgae throughout the Bay of Islands. Four new isolates of O. siamensis all produced palytoxin-like compounds (LC–MS analysis). Also isolated from Northland macroalgae were Amphidinium thermaeum (non-toxic by i.p. injection of mice) and Gambierdiscus cf. yasumotoi, with the latter producing a putative maitotoxin analogue (MTX-3) as determined by LC–MS.     

Response of epilithic diatom communities to environmental gradients along an Ecuadorian Andean River

Epilithic diatom communities are particularly suitable for the evaluation of freshwater quality. In Ecuador, however, no water quality index includes this biotic parameter. This work is the first attempt in the country to determine the composition of epilithic diatom communities associated with different degrees of eutrophication. This was accomplished by measuring physical, chemical and microbiological variables at five sampling sites along the Pita River, Ecuador, from August to December 2016. The results indicate a clear gradient of eutrophication from sampling sites P1 (good water) in the upper reaches to P5 (bad water) located in the lower reaches. Concerning diatom analyses, the results indicated a high diversity for tropical areas in terms of species richness, varying from S = 34 in headwaters to S = 42 downstream. Moreover, the results obtained suggest a lack of concordance with the trophic values given to some of the epilithic diatoms in the literature. There were also species that seem to be sensitive to downstream nutrient increases that were not considered as bioindicators in previous studies. We concluded that the trophic values of diatom species available in the scientific literature are not directly applicable to their sites in the Pita River. Hence, it is necessary to establish a trophic diatom index for the Andean region of Ecuador.     

Taxonomic diversity and community structure of arbuscular mycorrhizal fungi (Phylum Glomeromycota) in three maritime sand dunes in Santa Catarina state,south Brazil

Community structure and species richness of arbuscular mycorrhizal fungi (Phylum Glomeromycota) were studied in sand dune sites at Itapiruba (southern), Joaquina (intermediate) and Praia Grande (northern) beaches along the coast of the state of Santa Catarina, Brazil. In each site, a 20 × 20 m plot was established and 20 soil samples collected in a regular grid pattern. Fungal spores were extracted from each sample, counted and identified to species level. A total of 25 species were recovered belonging to seven genera and five families in the Glomeromycota. Gigaspora albida and Acaulospora scrobiculata occurred in >50 % of samples at all three sites. Other common species whose sample frequency was >50 % in one or two sites were Scutellospora weresubiae, Scutellospora cerradensis and Racocetra gregaria, while the remaining majority of species were detected in <25 % of samples within a given site. Dune sites could be differentiated based on the higher frequency of occurrence of S. cerradensis and Acaulospora morrowiae in Itapiruba, S. weresubiae in Joaquina, and Scutellospora hawaiiensis in Praia Grande. No differences across sites were observed for species richness and total spore numbers, the latter averaging from 28.8 to 31.8 spores per 100 ml soil. Shannon diversity was significantly higher in Praia Grande compared to the other two sites. Differences in the relative spore abundance of genera among dunes were detected only for Scutellospora, which was significantly more abundant in the Joaquina beach. Community structure, as depicted by species rank/log abundance graphs, was not significantly different between areas according to the Kolmogorov–Smirnov two-sample test. Species accumulation curves demonstrated that 13 samples were enough to detect 90 % of all species. Overall, sand dune systems share similar arbuscular mycorrhizal fungal communities despite being geographically distant (150 km) from each other.     

Light thresholds derived from seagrass loss in the coastal zone of the northern Great Barrier Reef,Australia

There is a world-wide trend for deteriorating water quality and light levels in the coastal zone, and this has been linked to declines in seagrass abundance. Localized management of seagrass meadow health requires that water quality guidelines for meeting seagrass growth requirements are available. Tropical seagrass meadows are diverse and can be highly dynamic and we have used this dynamism to identify light thresholds in multi-specific meadows dominated by Halodule uninervis in the northern Great Barrier Reef, Australia. Seagrass cover was measured at ∼3 month intervals from 2008 to 2011 at three sites: Magnetic Island (MI) Dunk Island (DI) and Green Island (GI). Photosynthetically active radiation was continuously measured within the seagrass canopy, and three light metrics were derived. Complete seagrass loss occurred at MI and DI and at these sites changes in seagrass cover were correlated with the three light metrics. Mean daily irradiance (I_d) above 5 and 8.4 mol m⁻² d⁻¹ was associated with gains in seagrass at MI and DI, however a significant correlation (R = 0.649, p < 0.05) only occurred at MI. The second metric, percent of days below 3 mol m⁻² d⁻¹, correlated the most strongly (MI, R = −0.714, p < 0.01 and DI, R = −0.859, p = <0.001) with change in seagrass cover with 16–18% of days below 3 mol m⁻² d⁻¹ being associated with more than 50% seagrass loss. The third metric, the number of hours of light saturated irradiance (H_sat) was calculated using literature-derived data on saturating irradiance (E_k). H_sat correlated well (R = 0.686, p < 0.01; and DI, R = 0.704, p < 0.05) with change in seagrass abundance, and was very consistent between the two sites as 4 H_sat was associated with increases in seagrass abundance at both sites, and less than 4 H_sat with more than 50% loss. At the third site (GI), small seasonal losses of seagrass quickly recovered during the growth season and the light metrics did not correlate (p > 0.05) with change in percent cover, except for I_d which was always high, but correlated with change in seagrass cover. Although distinct light thresholds were observed, the departure from threshold values was also important. For example, light levels that are well below the thresholds resulted in more severe loss of seagrass than those just below the threshold. Environmental managers aiming to achieve optimal seagrass growth conditions can use these threshold light metrics as guidelines; however, other environmental conditions, including seasonally varying temperature and nutrient availability, will influence seagrass responses above and below these thresholds.     

A 1700-year Athrotaxis selaginoides tree-ring width chronology from southeastern Australia

Few Southern Hemisphere tree-ring chronologies exceed 1000 years in length. We present a ca. 1700 years of indexed values for the long-lived conifer Athrotaxis selaginoides at Cradle Mt in southeastern Australia and compare it with the only other published millennial-plus length tree-ring chronology for Australia: the nearby Mt Read Lagarostrobos franklinii. We use simple correlation function and pointer year analyses to compare the climate responses of the two species (temperature, precipitation and growing degree days). Both chronologies show accelerated growth at their modern ends, but this growth acceleration is not synchronous, beginning approximately a quarter of a century earlier at the Cradle Mt site. This discrepancy may highlight the relevance of chronology composition and/or physiological differences in the species. Although the seasonality of the climatic responses of the two species is similar, that of A. selaginoides is generally weaker than that of L. franklinii. Somewhat paradoxically, the only pointer years in common between the chronologies are 1898 and 1908 CE. The periods from 600 to 900 CE and ∼1200–1450 CE are conspicuous for their absence of positive pointer years while no negative pointer years occur for either site from ∼1200–1350 CE. It is possible that differing patterns of pointer years can be partially explained by a peak in establishment from ∼1150–1850 CE at the Mt Read L. franklinii site compared to continuous establishment at Cradle Mt. Although statistically significant and time-stable climate responses for the A. selaginoides chronology are too weak to base a single-chronology climate reconstruction on, the long chronology will likely make an important contribution to future multi-proxy temperature reconstructions for southeastern Australia.