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Diarrhetic shellfish toxins (DSTs) are produced by the marine dinoflagellate, Dinophysis, as well as select species of benthic Prorocentrum. The DSTs can bioaccumulate in shellfish and cause gastrointestinal illness when humans consume high levels of this toxin. Although not routinely monitored throughout the U.S., recent studies in Washington, Texas, and New York suggest DSTs may be widespread throughout U.S. coastal waters. This study describes a four-year time series (2013-2016) of Dinophysis concentration and DST level in California mussels (Mytilus californianus) from Santa Cruz Municipal Wharf (SCMW) in Monterey Bay, California. Results show a maximum Dinophysis concentration of 9404 cells/L during this study and suggest Dinophysis persists as a member of the background phytoplankton community throughout the year. In California mussels, DSTs were found at persistent low levels throughout the course of this study, and exceeded the FDA guidance level of 160 ng/g 19 out of 192 weeks sampled. Concentrations of Dinophysis alone are a positive but weak predictor of DST level in California mussels, and basic environmental variables (temperature, salinity, and nutrients) do not sufficiently explain variation in Dinophysis concentration at SCMW. This study demonstrates that Dinophysis in Monterey Bay are producing DSTs that accumulate in local shellfish throughout the year, occasionally reaching levels of concern.

Due to the increasing prevalence of Dinophysis spp. and their toxins on every US coast in recent years, the need to identify and monitor for problematic Dinophysis populations has become apparent. Here, we present morphological analyses, using light and scanning electron microscopy, and rDNA sequence analysis, using a ~2-kb sequence of ribosomal ITS1, 5.8S, ITS2, and LSU DNA, of Dinophysis collected in mid-Atlantic estuarine and coastal waters from Virginia to New Jersey to better characterize local populations. In addition, we analyzed for diarrhetic shellfish poisoning (DSP) toxins in water and shellfish samples collected during blooms using liquid-chromatography tandem mass spectrometry and an in vitro protein phosphatase inhibition assay and compared this data to a toxin profile generated from a mid-Atlantic Dinophysis culture. Three distinct morphospecies were documented in mid-Atlantic surface waters: D. acuminata, D. norvegica, and a "small Dinophysis sp." that was morphologically distinct based on multivariate analysis of morphometric data but was genetically consistent with D. acuminata. While mid-Atlantic D. acuminata could not be distinguished from the other species in the D. acuminata-complex (D. ovum from the Gulf of Mexico and D. sacculus from the western Mediterranean Sea) using the molecular markers chosen, it could be distinguished based on morphometrics. Okadaic acid, dinophysistoxin 1, and pectenotoxin 2 were found in filtered water and shellfish samples during Dinophysis blooms in the mid-Atlantic region, as well as in a locally isolated D. acuminata culture. However, DSP toxins exceeded regulatory guidance concentrations only a few times during the study period and only in noncommercial shellfish samples.

The regional circulation in the northwestern Gulf of Mexico during late spring-summer is modulated by upwelling-favorable winds that can cause coastal upwelling in the western region and by freshwater inputs from the Mississippi-Atchafalaya Rivers. Spatial variability and temporal dynamics of phytoplankton community composition were examined during two upwelling-favorable periods using data obtained with an Imaging FlowCytobot (IFCB) from two cruises on the Texas-Louisiana shelf in June 2013 and 2014 and from the Texas Observatory for Algal Succession Time series (TOAST) at Port Aransas (Texas). Phytoplankton spatial distributions were determined by the influence of upwelling and river discharged waters. In the 2013 cruise, upwelling was detected in a large portion of the western region and the phytoplankton assemblages were dominated by diatoms, mostly chain-forming taxa. As revealed by the TOAST time series, the upwelling onset caused a dramatic increase in diatom carbon biomass. In the areas not affected by upwelling, variation in the river plume distribution that resulted from the circulation and the different discharge magnitudes for each year influenced the spatial distributions of the phytoplankton community composition. Dinoflagellates and other flagellated taxa were notably dominant during the 2013 cruise, whereas both diatoms and flagellated groups dominated the assemblages during the 2014 cruise. High stratification promoted by freshwater input, notably higher during 2013 than 2014, likely favored the dominance of flagellated groups in 2013. This study provides evidence of the influence of coastal upwelling in the phytoplankton community of the northwestern Gulf of Mexico and contributes to the knowledge of the drivers of community composition in this high-productivity area., This research was supported by grants from a Marie Curie international outgoing fellowship (GA-302562) from the European Community to S.A., and NOAA/ECOHAB (NA09NOS4780196) and NOAA/PCMHAB (NA15NOS4780173) to L.C. We thank L. Harred for collecting the IFCB data during the cruises. We extend our gratitude to the crew of the R/V Manta for their assistance on the cruises, and to S. DiMarco for providing the environmental data from his NOAA/MCH cruises (NA09NOS4780208). We thank E. Buskey and the Mission-Aransas NERR program, the NOAA/OAR/ESRL PSD, the National Data Buoy Center and the Texas Automated Buoy System, and the U.S. Army Corps of Engineers for providing data of Port Aransas Ship Channel station, satellite SST, wind and water temperature, and freshwater discharge, respectively, and R. Mooney for technical assistance with the IFCB at TOAST.

The cosmopolitan, species-rich diatom genus Pseudo-nitzschia represents a good system for the study of speciation, evolution and diversity. Understanding elements linked to population dynamics and life cycle regulation for these species is of particular importance in view of their ability to produce the toxin domoic acid and cause harmful blooms. Pseudo-nitzschia multistriata, one of the toxic species that represents a model for the study of life cycle related questions, is the only diatom for which a sex determination mechanism has been described. Populations in the Gulf of Naples (Mediterranean Sea), can share four different allelic variants (A, M, B, N) of the mating type determination region, and one of them (A) is responsible for the determination of the mating type + (MT+), defined by the MT+ restricted expression of the gene MRP3. Here, we analysed the sex determination genomic region in three new strains isolated from the Gulf of Mexico and compared it to the alleles previously described in the Mediterranean strains. We first show that these geographically distant strains of P. multistriata belong to different populations but can interbreed. Next, we show that the two populations share an overall similar structure of the genomic locus although differences can be seen in the polymorphic regions upstream of MRP3. In strain P4-C1, we amplified and sequenced an allele (M) identical to one of those previously characterized in the Mediterranean strains. In the other two strains, P4-C2 and P4-C5, we identified three new alleles, which we named A2, B2 and N2. P4-C2 and P4-C5 are heterozygous and share the common allele A2 linked to the monoallelic expression of the MT+ specific sex determining gene MRP3. Our results expand information on the global distribution of P. multistriata and on the level of conservation of the sex determination region in different populations. The definition of the extent of intra- and inter-specific conservation of this region would be a relevant addition to our understanding of Pseudo-nitzschia diversity and evolution.

Mixotrophic Dinophysis species threaten human health and coastal economies through the production of toxins which cause diarrhetic shellfish poisoning (DSP) in humans. Novel blooms of Dinophysis acuminata and Dinophysis ovum have occurred in North American waters in recent decades, resulting in the closure of shellfish harvesting. Understanding the ecology of Dinophysis species and their prey is essential to predicting and mitigating the impact of blooms of these dinoflagellates. The growth response of two new isolates of Dinophysis species, one isolate of Mesodinium rubrum, and two strains of Teleaulax amphioxeia were evaluated at a range of temperature, salinity, and irradiance treatments to identify possible environmental drivers of Dinophysis blooms in the Gulf of Mexico. Results showed optimal growth of T. amphioxeia and M. rubrum at 24 °C, salinity 30 – 34, and irradiances between 300 and 400 µmol quanta m − 2 s − 1. Optimal Dinophysis growth was observed at salinity 22 and temperatures between 18 and 24 °C. Mesodinium and both Dinophysis responded differently to experimental treatments, which may be due to the suitability of prey and different handling of kleptochloroplasts. Dinophysis bloom onset may be initiated by warming surface waters between winter and spring in the Gulf of Mexico. Toxin profiles for these two North American isolates were distinct; Dinophysis acuminata produced okadaic acid, dinophysistoxin-1, and pectenotoxin-2 while D. ovum produced only okadaic acid. Toxin per cell for D. ovum was two orders of magnitude greater than D. acuminata. Phylogenies based on the cox1 and cob genes did not distinguish these two Dinophysis species within the D. acuminata complex.

Prochlorococcus is a ubiquitous marine oxyphotobacterium characterized by the presence of DV-chl a and b. In addition, the type strain Prochlorococcus marinus Chisholm et al. CCMP 1375 (or SS120), an isolate from the Sargasso Sea, contains low levels of an unusual phycoerythrin. Until now, it has been unclear if phycoerythrin occurs randomly within this systematic group and if the molecular characteristics of this phycoerythrin are restricted to this single strain. Here, we show that two additional Prochlorococcus strains from the Pacific Ocean also contain similar low levels of phycoerythrin. DNA sequence and phylogenetic analyses demonstrated that this phycoerythrin is very similar to the phycoerythrin of P. marinus SS120 and differs from the classic cyanobacterial phycoerythrins. In contrast, a third isolate from the Arabian Sea lacks phycoerythrin. Based on the DV-chl b:a ratio and 16S rRNA sequence data, we classify the two Pacific phycoerythrin-containing isolates as low-light-adapted strains and the Arabian Sea isolate as a high-light-adapted strain. Thus, we provide further evidence to link the physiology of an individual genotype and the presence or absence of functional phycoerythrin genes within the genus Prochlorococcus.

Importance Patient-reported outcome (PRO) data from clinical trials can provide valuable evidence to inform shared decision making, labeling claims, clinical guidelines, and health policy; however, the PRO content of clinical trial protocols is often suboptimal. The SPIRIT (Standard Protocol Items: Recommendations for Interventional Trials) statement was published in 2013 and aims to improve the completeness of trial protocols by providing evidence-based recommendations for the minimum set of items to be addressed, but it does not provide PRO-specific guidance. Objective To develop international, consensus-based, PRO-specific protocol guidance (the SPIRIT-PRO Extension). Design, Setting, and Participants The SPIRIT-PRO Extension was developed following the Enhancing Quality and Transparency of Health Research (EQUATOR) Network’s methodological framework for guideline development. This included (1) a systematic review of existing PRO-specific protocol guidance to generate a list of potential PRO-specific protocol items (published in 2014); (2) refinements to the list and removal of duplicate items by the International Society for Quality of Life Research (ISOQOL) Protocol Checklist Taskforce; (3) an international stakeholder survey of clinical trial research personnel, PRO methodologists, health economists, psychometricians, patient advocates, funders, industry representatives, journal editors, policy makers, ethicists, and researchers responsible for evidence synthesis (distributed by 38 international partner organizations in October 2016); (4) an international Delphi exercise (n = 137 invited; October 2016 to February 2017); and (5) consensus meeting (n = 30 invited; May 2017). Prior to voting, consensus meeting participants were informed of the results of the Delphi exercise and given data from structured reviews evaluating the PRO protocol content of 3 defined samples of trial protocols. Results The systematic review identified 162 PRO-specific protocol recommendations from 54 sources. The ISOQOL Taskforce (n = 21) reduced this to 56 items, which were considered by 138 international stakeholder survey participants and 99 Delphi panelists. The final wording of the SPIRIT-PRO Extension was agreed on at a consensus meeting (n = 29 participants) and reviewed by external group of experts during a consultation period. Eleven extensions and 5 elaborations to the SPIRIT 2013 checklist were recommended for inclusion in clinical trial protocols in which PROs are a primary or key secondary outcome. Extension items focused on PRO-specific issues relating to the trial rationale, objectives, eligibility criteria, concepts used to evaluate the intervention, time points for assessment, PRO instrument selection and measurement properties, data collection plan, translation to other languages, proxy completion, strategies to minimize missing data, and whether PRO data will be monitored during the study to inform clinical care. Conclusions and Relevance The SPIRIT-PRO guidelines provide recommendations for items that should be addressed and included in clinical trial protocols in which PROs are a primary or key secondary outcome. Improved design of clinical trials including PROs could help ensure high-quality data that may inform patient-centered care.

Ectopic pregnancies complicate 1–2 pregnancies and are a leading cause of maternal death. An effective oral drug therapy that replaces surgery might make its treatment safer, cheaper, simpler and therefore more widely accessible. The only current medical treatment offered to women is intramuscular methotrexate, but this only reliably resolves smaller ectopic pregnancies. As such, many ectopic pregnancies require surgical excision. We show that vinorelbine, an orally available chemotherapeutic agent, potently induced placental cell death but did not harm fertility in mice. Vinorelbine was 100–1000 times more potent than methotrexate in inducing placental cell death in vitro, and more potent than combination methotrexate and gefitinib (another proposed treatment for ectopic pregnancy being evaluated in phase III trials). Mechanistically, it caused microtubule condensation, blocked mitosis and activated the apoptosis cascade in placental cells. Vinorelbine was more efficacious than methotrexate ± gefitinib in reducing the volume of placental cell tumors xenografted subcutaneously in SCID mice. Mice exposed to vinorelbine and allowed to breed, following a four week washout period, displayed normal fertility, however long-term fertility was not assessed. Human Fallopian tubes treated with vinorelbine did not exhibit up-regulation of apoptosis molecules. Our findings show that placental cells appear sensitive to vinorelbine and it has potential as a tablet-only approach to treat ectopic pregnancy., Highlights • We have identified vinorelbine, a well tolerated chemotherapeutic, as a potential therapeutic for ectopic pregnancy. • Vinorelbine potently reduced placental cell viability in-vitro and in-vivo and proved more efficacious than the current medical therapeutic. • Vinorelbine did not impact upon subsequent fertility in a mouse breeding model. Ectopic pregnancy occurs when a conceptus implants outside of the womb. They are life threating and are a leading cause of maternal death in early pregnancy. Currently, the large majority of ectopic pregnancies are removed surgical, partly owing to the limited efficacy of the only available medical therapeutic, methotrexate. Here, we have identified a potential new medical therapeutic for ectopic pregnancy, vinorelbine. Vinorelbine, which is orally available chemotherapeutic, reduced placental cell viability both in-vitro and in-vivo and did not impact upon mice fertility in a breeding model. As such, vinorelbine may be an efficacious treatment for ectopic pregnancy and further human studies into its application are warranted.

Ectopic pregnancy (EP), where an embryo implants outside of the normal uterine cavity, occurs in 1%–2% of all pregnancies. It is termed heterotopic if there is also a concurrent intrauterine pregnancy. 95%–98% of EPs occur within the Fallopian tube where placental invasion of maternal vasculature can cause life-threatening hemorrhage. Factors that alter or impede embryo transport can increase the risk of developing EP. Detection is largely by transvaginal ultrasound and, where there is diagnostic uncertainty, serial serum β-hCG monitoring. EP can be managed surgically, medically or expectantly. Further improvements in diagnostics and medical management are required to improve patient outcomes.

Harmful algal blooms caused by Karenia brevis result in large fish kills, human respiratory irritation, and shellfishing closures in affected areas. Most previous work on bloom formation in the Gulf of Mexico has focused on the west coast of Florida. To investigate the origin of bloom-forming cells along the coast of Texas, potential distributions of cells during two bloom years (2009, 2011) and one non-bloom year (2010) were examined using a spatially explicit, individual-based model of K. brevis. The model incorporates a previously developed model of dinoflagellate vertical migration and utilizes observed data (field samples of cell concentrations, photosynthetically active radiation) and modeled environmental output (salinity, temperature, current velocities) from a hydrodynamic model. Running the model in reverse showed that cells near the coast of Texas during early fall originate from the southern Gulf of Mexico in bloom years and from the northern Gulf of Mexico in the non-bloom year for the three years studied. Identification of a southern origin for bloom-forming cells provides a target area for increased sampling in order to provide early warning of potentially harmful algal blooms of K. brevis.

In this article, three librarians from Santa Fe College, the community college winner of the 2015 ACRL Excellence in Academic Libraries Award, share their successful work in information literacy instruction. While the ACRL award is given for overall achievement, this article focuses upon their creative and varied approaches to instruction, incorporating a wide blend of strategies to reach students across the college. Beginning with targeted approaches to virtual instruction, including broad use of the flipped classroom model, and building in active learning techniques, the librarians at Santa Fe College have built an instruction program that could serve as a model to community college libraries across the country.

To investigate the response of a subtropical coastal phytoplankton community to tropical cyclones, we utilized high temporal resolution (hours) data from the Imaging FlowCytobot (IFCB) deployed in the Gulf of Mexico. In 2010, four tropical cyclones (two during June–July and two during September) struck the area. Passage of the cyclones produced two major impacts: (1) storm surges and strong onshore winds, and (2) heavy rains producing substantial freshwater discharge and decreased salinity. The phytoplankton community showed a rapid response to the passage of the cyclones with increased abundance. Using principal component analysis, responses during storm surges and after freshwater discharges were distinguished and considerable changes in community composition were revealed. Responses to storm surges were characterized by an increase of diatoms. Freshwater discharges triggered increases in dinoflagellates and other flagellates (prasinophytes, euglenophytes, and cryptophytes) relative to the other groups. In June–July, substantial increases in diatoms were also observed. The response to freshwater discharge during this period was dominated by a diatom (Thalassiosira), whereas in September the response was dominated by two dinoflagellates (Akashiwo sanguinea and Polykrikos hartmannii). Observed abundances of these three taxa were the highest recorded from the IFCB time series. The short-term responses of the phytoplankton community revealed here emphasize the need for high temporal resolution sampling to fully capture the effects of tropical cyclones. Given that extreme storms are predicted to increase with future climate change, the taxonomic resolution of the IFCB is also valuable for detecting taxa-specific responses, which can have implications for ecosystem functioning.

Blooms of Dinophysis ovum and Mesodinium spp. have been observed in the Gulf of Mexico since 2007 using the Imaging FlowCytobot technology. Bloom dynamics of these two organisms in conjunction with ancillary environmental data for a 5-year period were analyzed to identify the conditions necessary for bloom initiation or presence with the goal of predicting future blooms of D. ovum. Using time-series analysis, we observed a positive time-lagged correlation between the two organisms in each year when both were present, which suggests that the presence of Mesodinium may be useful as a leading indicator for a D. ovum bloom. Although in some cases D. ovum and Mesodinium co-occurred, no strong predator– prey relationship was observed. We identified a narrow range of temperature and salinity that could be necessary for bloom initiation of D. ovum and Mesodinium in the Gulf of Mexico. Analysis of images over the time series revealed a wide range in the size of Mesodinium cells, which suggests that species other than M. rubrum may be present in the Gulf of Mexico. Based on the occurrence of a D. ovum bloom preceded by low abundances of Mesodinium, we suggest that D. ovum is able to utilize ciliates other than M. rubrum as prey. Our observations indicate that environmental conditions, as well as Mesodinium abundance and species composition, can affect initiation, presence or abundance of D. ovum and thus may help in the prediction of future blooms.

Increasing atmospheric CO2 is promoting ocean acidification and higher global temperatures and has been suggested as a possible factor for shifts in marine phytoplankton composition to more harmful species. Karenia brevis is the major harmful algal species in the Gulf of Mexico producing potent neurotoxins known as brevetoxins. We examined how changes in ocean inorganic carbon chemistry associated with pre-industrial (250 ppm), recent (350 ppm), and predicted at 2100 (1000 ppm) pCO2 levels and increased temperature affect growth rates and brevetoxin production in K. brevis. At the predicted pCO2 levels for 2100, growth rate of K. brevis Wilson clone increased substantially by 46% at 25 °C (0.43 ± 0.01 d−1) compared to recent and pre-industrial levels (0.29 ± 0.01 d−1). Growth rates also increased for a low brevetoxin-producing clone, SP1, from 0.24 ± 0.02 d−1 at lower pCO2 levels to 0.33 ± 0.003 d−1 at a pCO2 of 1000 ppm. When grown at a higher temperature (30 °C), growth rates for the Wilson clone significantly decreased at all three pCO2 by approximately 30%. However, even at the higher temperature, K. brevis growth rate significantly increased by 30% (0.30 ± 0.01 d−1) at the 1000 ppm CO2 level when compared to recent and pre-industrial CO2 levels (0.21 ± 0.01 d−1). Although K. brevis growth rate decreased at higher temperatures, the growth rate at pCO2 level of 1000 and 30 °C was slightly higher than at current conditions (pCO2 level of 350 and 25 °C). Modification of pCO2 levels and temperatures did not have an effect on total brevetoxin production or brevetoxin profiles in either clone examined. Due to the increased growth rate, total brevetoxin production was significantly higher at the pCO2 level of 1000. Finally, from these results we conclude there is no connection between growth rate and brevetoxin per cell. Although neither pCO2 nor temperature influenced brevetoxin production per cell, we suggest that under predicted future climate conditions K. brevis blooms have the potential to produce higher cell concentrations and increased brevetoxin concentrations, which will pose an increased risk for ecosystem and human health.

Background and aims Little evidence exists for the effectiveness of intervention for older adolescents and young adults with language disorders, particularly for those over 16 years. This study involves college-aged students aged 16–19 years with Language Disorder and Word-Finding Difficulties and investigates whether progress in word finding following 1:1 semantic intervention is greater than progress during a baseline period as measured by a standardised test. Methods Twenty-five college-aged students (20 males and 5 females) with Language Disorder and Word-Finding Difficulties (aged 16;4–18;4 years) participated in a single baseline design study comparing progress on the Test of Adolescent and Adult Word Finding during an intervention period with progress during a baseline period of equal length. Intervention was focused on semantics of different words from those in the Test of Adolescent and Adult Word Finding and was delivered 1:1 by the participants’ usual Speech and Language Therapist, 30 minutes per week, for eight weeks. Results The participants made significant progress in raw score on the Test of Adolescent and Adult Word Finding during both the baseline ( d = 1.4) and intervention ( d = 2.5) periods, but progress during the intervention period was significantly greater than during the baseline period ( d = 1.16). Individual data showed reliable change for five participants during the baseline period and for 20 participants during the intervention period. At the start of the study, all participants had standard scores below 85, but after intervention, 10/25 participants scored above 85. Conclusions Four hours of semantic intervention led to significantly greater gains on a standardised test of word finding than during a baseline period of equal length in 16–19 year olds with Language Disorder and Word-Finding Difficulties. The words in the standardised test had not been included in the intervention, indicating generalised gains. Implications This study shows that intervention (at least for Word-Finding Difficulties) can be effective for this older age group of college-aged students with Language Disorder and therefore the effectiveness of Speech and Language Therapy services for this age group in a wider range of areas of language should also be investigated.

[1] Atmospheric mixing ratios and seawater concentrations of bromoform (CHBr3), dibromomethane (CH2Br2), and other brominated very short-lived substances (BrVSLS) were measured during five cruises from 1994 to 2010. These cruises were conducted over large latitudinal (62°N–60°S) and longitudinal transects (11°W–86°W) in the Atlantic Ocean. Elevated seawater concentrations of CHBr3 and CH2Br2 were often observed in regions where chlorophyll a concentrations were also elevated, which suggests biogeochemical processes associated with photosynthetic biomass may be related to CHBr3 and CH2Br2 production. Our results suggest that, at least in the open ocean, several phytoplankton taxa may contribute to the production of these trace gases. While observed correlations between CHBr3 and CH2Br2 in different regions are usually interpreted as common sources for these compounds, results in this study suggest different biogeochemical processes may contribute separately to the production of these trace gases. Heterotrophic bacterial abundance was significantly correlated with CH2Br2, but not with CHBr3, which suggests the biogeochemical processes associated with heterotrophic bacteria may be related to CH2Br2 in seawater but probably not to CHBr3. In general, the Atlantic Ocean is a net source for CHBr3 and CH2Br2, except for a few locations where these trace gases were undersaturated in seawater. Assuming fluxes measured in the Atlantic open ocean are globally representative, the resulting extrapolated, global open-ocean annual net sea-to-air fluxes calculated from data from the five cruises was estimated at 0.24–3.80 Gmol Br yr−1 for CHBr3 and 0.11–0.77 Gmol Br yr−1 for CH2Br2.

[1] Seawater concentrations and distributions of brominated very short-lived substances (BrVSLS), including bromoform (CHBr3), dibromomethane (CH2Br2), bromodichloromethane (CHBrCl2), chlorodibromomethane (CHClBr2), were measured in the upper water column (5–750 m) in the eastern Pacific. Inorganic nutrient, pigment concentrations, and picoplankton cell counts were measured to determine biogeochemical factors that affect the production and distribution of these BrVSLS. Elevated concentrations of BrVSLS were observed in coastal and tropical seawater. Concentration maxima for CHBr3, CH2Br2, and CHClBr2 were observed below the mixed layer, near the subsurface chlorophyll a maxima, which suggest BrVSLS production may be related to photosynthetic biomass production. Our results also suggest that heterotrophic bacteria may also contribute to CH2Br2 and CHBrCl2 production in the water column. The maximum CHBrCl2 concentration was observed at a depth much deeper than the euphotic zone, which suggests sources other than photosynthetic biomass. Elevated CHBrCl2 concentrations in deeper waters were coincident with elevated CHCl3 concentrations, which may be an evidence for successive chlorine substitution of CHBr3 in deeper and older water masses.

Canola oil-based diets have been shown to reduce plasma cholesterol levels in comparison with diets containing higher levels of saturated fatty acids. Consumption of canola oil also influences biological functions that affect various other biomarkers of disease risk. Previous reviews have focused on the health effects of individual components of canola oil. Here, the objective is to address the health effects of intact canola oil, as this has immediate practical implications for consumers, nutritionists, and others deciding which oil to consume or recommend. A literature search was conducted to examine the effects of canola oil consumption on coronary heart disease, insulin sensitivity, lipid peroxidation, inflammation, energy metabolism, and cancer cell growth. Data reveal substantial reductions in total cholesterol and low-density lipoprotein cholesterol, as well as other positive actions, including increased tocopherol levels and improved insulin sensitivity, compared with consumption of other dietary fat sources. In summary, growing scientific evidence supports the use of canola oil, beyond its beneficial actions on circulating lipid levels, as a health-promoting component of the diet.

In the mammalian ovary, oocytes are contained within ovarian follicles. These consist in an oocyte surrounded by supporting cells: an inner layer of granulosa cells and an outer layer of thecal cells separated by a basal lamina. At any one time, a developing cohort of follicles exists, from which only a small species-specific number are selected for continued development towards ovulation, with the remainder dying by follicular atresia. Here, we use in vitro methods to study interactions between two follicles in culture (follicle co-cultures). We show that, when two individual follicles are grown together in culture, cells and cellular processes migrate from the outer thecal layer of one follicle to the thecal layer of the other co-cultured follicle. These cells are identified as a mixed population containing primarily endothelial but also neuronal cells. Both are able to migrate through the ovarian interstitum, making contact with the basal lamina of other follicles and with similar cells from these other follicles. Networks of such cells might be involved in interfollicular communication and in the coordination of follicle selection for ovulation.

The dinoflagellate Karenia brevis is the major harmful algal bloom (HAB) species in the Gulf of Mexico. Given that the rapid appearance of K. brevis cannot be explained by plankton growth alone, advection is likely important in bloom initiation. Forward- and backward-moving numerical surface drifters were employed in a numerical model of the Texas–Louisiana shelf to help determine the basic physical mechanisms explaining sporadic interannual occurrences of K. brevis along the Texas coastline. Results from data analysis from the area show that HAB events occur in years in which there are weaker mean downcoast, along-shore wind speeds. The drifter experiments suggest that southern waters play a role in HAB event initiation, providing an offshore source of cells at the end of summer. As winds switch from upcoast to downcoast in early fall, offshore populations of K. brevis are swept southward by wind-driven currents in years with strong downcoast winds. However, when the downshore wind is weak, shoreward Ekman transport creates a convergent flow near the coast that allows cells to concentrate and initiate a bloom.

Blooms of Karenia brevis, the major bloom-forming dinoflagellate in the Gulf of Mexico, are thought to originate in the eastern Gulf. Single-cell polymerase chain reaction and five microsatellites were used to obtain genotypes for > 1800 cells from 38 samples collected from six bloom events. A consistent pattern of genetic divergence between blooms from Florida and Texas was not detected, which supports the hypothesis of a common origin for blooms of K. brevis in the Gulf of Mexico.

Monitoring programs for harmful algal blooms (HABs) typically rely on time-consuming manual methods for identification and enumeration of phytoplankton, which make it difficult to obtain results with sufficient temporal resolution for early warning. Continuous automated imaging-in-flow by the Imaging FlowCytobot (IFCB) deployed at Port Aransas, TX has provided early warnings of six HAB events. Here we describe the progress in automating this early warning system for blooms of Karenia brevis. In 2009, manual inspection of IFCB images in mid-August 2009 provided early warning for a Karenia bloom that developed in mid-September. Images from 2009 were used to develop an automated classifier that was employed in 2011. Successful implementation of automated file downloading, processing and image classification allowed results to be available within 4 h after collection and to be sent to state agency representatives by email for early warning of HABs. No human illness (neurotoxic shellfish poisoning) has resulted from these events. In contrast to the common assumption that Karenia blooms are near monospecific, post-bloom analysis of the time series revealed that Karenia cells comprised at most 60-75 % of the total microplankton.

A new planktonic species of Prorocentrum is described from the Gulf of Mexico. First observed with the Imaging FlowCytobot, Prorocentrum texanum sp. nov. was characterized using LM, SEM, and TEM along with sequencing of the SSU, LSU, and ITS ribosomal regions and the mitochondrial cob and cox1 regions. P. texanum sp. nov. is a round to oval bivalvate dinoflagellate, with a prominent anterior, serrated solid flange on periflagellar a platelet and an opposing short, flat flange on the h platelet. The periflagellar area consists of 10 platelets. Both left and right valves have shallow round depressions and two-sized valve pores. The anterior ejectosome pore pattern differs between the left and right valve in relation to the periflagellar area and margins. Ten to eleven rows of tangential ejectosome pores are present on each valve. P. texanum sp. nov. has two varieties which exhibit distinct morphotypes, one round to oval (var. texanum) and the other pointed (var. cuspidatum). P. texanum var. cuspidatum is morphologically similar to P. micans in surface markings, but is smaller, and has a serrated periflagellar flange, and is genetically distinct from P. micans. Cytologically, P. texanum has two parietal chlo-roplasts, each with a compound, interlamellar pyrenoid, trichocysts, fibrous vesicles that resemble mucocysts, pusules, V- to U-shaped posterior nucleus, golgi, and tubular mitochondria. No genetic difference was found between the two varieties in the five genes examined. Phylogenetic analysis of the SSU, LSU, and ITS ribosomal regions place P. texanum sp. nov. as a sister group to P. micans. One isolate of P. texanum var. texanum produces okadaic acid.

Karenia brevis is the major harmful bloom-forming dinoflagellate in the Gulf of Mexico yet little is known about the intraspecific genetic diversity of this species. Here we describe nine new microsatellite markers and, combined with nine previously described microsatellites, use them to genotype 40 cultured isolates of K. brevis. Genetic diversity identified from cultured isolates was compared with the genetic diversity identified from two field samples to assess how well the current cultures represent the field population. Thirty-nine unique haplotypes were identified from 40 cultured isolates of K. brevis using 18 microsatellite markers. Genetic diversity was similar between cultured isolates and the two field samples. The success of 18 microsatellite markers to distinguish individual isolates supports the use of microsatellites as a genetic tool for diagnostic identification of cultured isolates of K. brevis.

Objective: This pragmatic randomized study was conducted to compare the effectiveness of telephone and face-to-face counseling in influencing problematic gambling beliefs and behaviors. Method: Ninety-two participants from diverse ethnic backgrounds who had been affected by problem gambling were provided with psychological interventions delivered either by telephone or conventional face-to-face counseling over a 3-month period. Results: A significant overall time effect between pre- and postintervention assessments was found for total hours, money and proportion of income spent, and the attitudes and beliefs scores. There was no significant difference in effect size between the two groups. Conclusion: This study provides preliminary evidence that both face-to-face and telephone counseling interventions might be equally effective in terms of short-term clinical outcomes measured postintervention.

At present, canola meal is primarily streamlined into the animal feed market where it is a competitive animal feed source owing to its high protein value. Beyond animal feed lies a potential game-changer with regards to the value of canola meal, and its opportunity as a high quality food protein source. An economic and sustainable source of protein with high bioavailability and digestibility is essential to human health and well-being. Population pressures, ecological considerations, and production efficiency underscore the importance of highly bioavailable plant proteins, both for the developed and developing world. Despite decades of research, several technologies being developed, and products being brought to large scale production, there are still no commercially available canola protein products. The workshop entitled “Canola/Rapeseed Protein—Future Opportunities and Directions” that was held on 8 July 2015 during the 14th International Rapeseed Congress (IRC 2015) addressed the current situation and issues surrounding canola meal protein from the technological, nutritional, regulatory and genomics/breeding perspective. Discussions with participants and experts in the field helped to identify economic barriers and research gaps that need to be addressed in both the short and long term for the benefit of canola industry.

Brachidinium capitatum F. J. R. Taylor, typically considered a rare oceanic dinoflagellate, and one which has not been cultured, was observed at elevated abundances (up to 65 cells · mL(-1) ) at a coastal station in the western Gulf of Mexico in the fall of 2007. Continuous data from the Imaging FlowCytobot (IFCB) provided cell images that documented the bloom during 3 weeks in early November. Guided by IFCB observations, field collection permitted phylogenetic analysis and evaluation of the relationship between Brachidinium and Karenia. Sequences from SSU, LSU, internal transcribed spacer (ITS), and cox1 regions for B. capitatum were compared with five other species of Karenia; all B. capitatum sequences were unique but supported its placement within the Kareniaceae. From a total of 71,487 images, data on the timing and frequency of dividing cells was also obtained for B. capitatum, allowing the rate of division for B. capitatum to be estimated. The maximum daily growth rate estimate was 0.22 d(-1) . Images showed a range in morphological variability, with the position of the four major processes highly variable. The combination of morphological features similar to the genus Karenia and a phylogenetic analysis placing B. capitatum in the Karenia clade leads us to propose moving the genus Brachidinium into the Kareniaceae. However, the lack of agreement among individual gene phylogenies suggests that the inclusion of different genes and more members of the genus Karenia are necessary before a final determination regarding the validity of the genus Brachidinium can be made.

Buitenhuis, Erik et al., The smallest marine phytoplankton, collectively termed picophytoplankton, have been routinely enumerated by flow cytometry since the late 1980s during cruises throughout most of the world ocean. We compiled a database of 40 946 data points, with separate abundance entries for Prochlorococcus, Synechococcus and picoeukaryotes. We use average conversion factors for each of the three groups to convert the abundance data to carbon biomass. After gridding with 1° spacing, the database covers 2.4% of the ocean surface area, with the best data coverage in the North Atlantic, the South Pacific and North Indian basins, and at least some data in all other basins. The average picophytoplankton biomass is 12 ± 22 μg C l−1 or 1.9 g C m−2. We estimate a total global picophytoplankton biomass of 0.53–1.32 Pg C (17–39% Prochlorococcus, 12–15% Synechococcus and 49–69% picoeukaryotes), with an intermediate/best estimate of 0.74 Pg C. Future efforts in this area of research should focus on reporting calibrated cell size and collecting data in undersampled regions., We thank Claude Belzile, Jacques Neveux and Genevieve Tremblay for their comments on a draft manuscript, the EU (CarboChange, contract 264879) for financial support to ETB, and the Networks of Centres of Excellence of Canada-ArcticNet for financial support to MG.

Brachidinium capitatum F. J. R. Taylor, typically considered a rare oceanic dinoflagellate, and one which has not been cultured, was observed at elevated abundances (up to 65 cells · mL(-1) ) at a coastal station in the western Gulf of Mexico in the fall of 2007. Continuous data from the Imaging FlowCytobot (IFCB) provided cell images that documented the bloom during 3 weeks in early November. Guided by IFCB observations, field collection permitted phylogenetic analysis and evaluation of the relationship between Brachidinium and Karenia. Sequences from SSU, LSU, internal transcribed spacer (ITS), and cox1 regions for B. capitatum were compared with five other species of Karenia; all B. capitatum sequences were unique but supported its placement within the Kareniaceae. From a total of 71,487 images, data on the timing and frequency of dividing cells was also obtained for B. capitatum, allowing the rate of division for B. capitatum to be estimated. The maximum daily growth rate estimate was 0.22 d(-1) . Images showed a range in morphological variability, with the position of the four major processes highly variable. The combination of morphological features similar to the genus Karenia and a phylogenetic analysis placing B. capitatum in the Karenia clade leads us to propose moving the genus Brachidinium into the Kareniaceae. However, the lack of agreement among individual gene phylogenies suggests that the inclusion of different genes and more members of the genus Karenia are necessary before a final determination regarding the validity of the genus Brachidinium can be made.

Peer education is used as a health promotion strategy in a number of areas, including sexual health. Although peer education programmes have been around for some time, published systematic evaluations of youth sexual health peer education programmes are rare. This article discusses the advantages and disadvantages of youth sexual health peer education programmes, the importance of programme evaluation, and strategies for developing effective programme evaluation tools. The value of conducting both process (programme delivery) and outcome (programme impact) evaluation is examined as well as methods for conducting these forms of assessment. Considering the wide range of peer education programmes and the diversity of communities served, the article concludes that the creation of a single evaluation method may be an impossible task. To address this challenge, principles for effective programme evaluation are proposed with tools that can be tailored to the unique goals of specific sexual health organizations.

Uptake of inorganic and organic nitrogen (N) by phytoplankton and bacteria was inves- tigated during a mesocosm study conducted in Raunefjord, Norway in April 2005. One mesocosm was fertilized with nitrate and phosphate at a ratio of 16:1 and maintained in the light, while one un- amended light mesocosm served as a control. Dissolved nutrients, phytoplankton and bacterial bio- mass, and phytoplankton community composition were monitored throughout the 26 d experiment. Uptake of 15 N-labeled ammonium and nitrate, and dual-labeled ( 15 N and 13 C) urea and dissolved free amino acids (DFAA) was measured for phytoplankton and bacteria using 2 methods: size fractiona- tion into 0.2-0.8 and >0.8 μm size classes, and flow cytometric sorting based on chlorophyll autoflu- orescence. Prior to fertilization, dissolved inorganic N concentrations were low and comprised ~5% of total dissolved N. Added nitrate was completely utilized in the amended mesocosm within 10 d, stimulating a large bloom of colonial Phaeocystis pouchetii. Ammonium contributed over half of total measured N uptake by phytoplankton and bacteria in both enclosures, while nitrate and urea each supplied roughly 10 to 25%. Overall, DFAA were a negligible N source to phytoplankton but con- tributed 11% to total bacterial N uptake. Bacterial uptake represented a significant portion of total uptake of all N forms, especially urea and DFAA. Comparison of the 2 methods for measuring phyto- plankton versus bacterial uptake demonstrates how the use of 0.8 μm filters can lead to significant overestimation of phytoplankton N uptake due to the retention of bacterial biomass.

Nucleic acid sequence-based amplification (NASBA) is an isothermal method used to amplify RNA and has been used for clinical, environmental, and food testing applications. Quantification of RNA by realtime NASBA occurs by comparing time to positive (TTP) fluorescence values, similar to threshold cycle (Ct) values in PCR, of unknown samples to a standard curve of known RNA titers. Incorporation of an internal control RNA molecule (IC-RNA) has been used to increase precision and accuracy of real-time NASBA and also serves as an indicator of NASBA inhibition. A real-time IC-NASBA assay was developed targeting the ribulose-1,5-bisphosphate carboxylase-oxygenase (RuBisCO) large-subunit gene (rbcL )o f the harmful algal bloom (HAB) causing dinoflagellate Karenia mikimotoi. This assay is sensitive to one K. mikimotoi cell and 1 � 10

The dinoflagellate Karenia brevis is responsible for nearly annual red tides in the Gulf of Mexico that cause extensive marine mortalities and human illness due to the production of brevetoxins. Although the mechanisms regulating its bloom dynamics and toxicity have received considerable attention, investigation into these processes at the cellular and molecular level has only begun in earnest during the past decade. This review provides an overview of the recent advances in our understanding of the cellular and molecular biology on K. brevis. Several molecular resources developed for K. brevis, including cDNA and genomic DNA libraries, DNA microarrays, metagenomic libraries, and probes for population genetics, have revolutionized our ability to investigate fundamental questions about K. brevis biology. Two cellular processes have received particular attention, the vegetative cell cycle and vertical migration behavior, which are of key importance due to their roles in the development of both surface populations that constitute blooms and subsurface cell aggregations that may serve to initiate them. High throughput sequencing of cDNA libraries has provided the first glimpse of the gene repertoire in K. brevis, with approximately 12,000 unique genes identified to date. Phylogenomic analysis of these genes has revealed a high rate of horizontal gene transfer in K. brevis, which has resulted in a chimeric chloroplast through the selective retention of genes of red, green, and haptophyte origin, whose adaptive significance is not yet clear. Gene expression studies using DNA microarrays have demonstrated a prevalence of post-transcriptional gene regulation in K. brevis and led to the discovery of an unusual spliced leader trans-splicing mechanism. Among the trans-spliced gene transcripts are type I polyketide synthases (PKSs), implicated in brevetoxin biosynthesis, which are unique among type I PKSs in that each transcript encodes an individual catalytic domain, suggesting a novel gene structure in this dinoflagellate. Clone libraries of 16S ribosomal DNA sequences developed from bloom waters have unveiled the temporal and spatial complexity of the microbial soup that coexists with K. brevis and its active involvement in both bloom growth and termination processes. Finally, the development and application of population genetic markers has revealed a surprisingly high genetic diversity in K. brevis blooms, long assumed to consist of essentially clonal populations. With these foundations in place, our understanding of K. brevis bloom dynamics is likely to grow exponentially in the next few years.

A simple and effective protocol is described for multiplex polymerase chain reaction (PCR) amplification of single cells of Karenia brevis. The protocol requires minimum processing, avoids additions that might dilute target DNA template, and can be used on cells preserved in Lugol's iodine preservative. Destaining of Lugol's-preserved cells with sodium thiosulfate allowed successful amplification of single-copy, nuclear-encoded microsatellites in single cells of K. brevis that have been preserved for up to 6 years.

OBJECTIVE: To investigate whether gonadotropin-releasing hormone (GnRH) and GnRH receptor (GnRHR) are expressed at tubal ectopic pregnancy sites, and to study the potential role of GnRH signaling in regulating immortalized human trophoblast cell viability.DESIGN: Immunohistochemical and experimental studies.SETTING: Academic research laboratory.PATIENT(S): Fallopian tube implantation sites (n = 25) were collected from women with ectopic pregnancy. First-trimester human placenta biopsies (n = 5) were obtained from elective terminations of pregnancy.INTERVENTION(S): None.MAIN OUTCOME MEASURE(S): GnRH and GnRHR expression was examined by means of immunohistochemistry and histoscoring. Trophoblastic BeWo choriocarcinoma and immortalized extravillous trophoblast (HTR-8/SVneo) cell viability was examined by means of cell counting after incubation with GnRH and/or GnRH antagonist (Antide).RESULT(S): GnRH and GnRHR immunoreactivity was detected in cytotrophoblast, syncytiotrophoblast, and extravillous trophoblast in all women with tubal pregnancy. GnRH immunoreactivity was higher and GnRHR immunoreactivity lower in syncytiotrophoblast compared with cytotrophoblast. GnRH and GnRHR immunoreactivity was detected in adjacent fallopian tube epithelium. Whereas neither GnRH nor Antide altered HTR-8/SVneo cell viability, treatment with GnRH significantly increased the overall cell viability of BeWo cells at 48 and 72 hours, and these effects were abolished by pretreatment with Antide.CONCLUSION(S): GnRH and GnRHR are expressed in trophoblast cell populations and fallopian tube epithelium at tubal ectopic pregnancy sites. GnRH increases BeWo cell viability, an effect mediated by the GnRHR. Further work is required to investigate the potential role of GnRH signaling in ectopic pregnancy.

Penicillin binding proteins (PBPs) are the target of numerous antimicrobial agents that disrupt bacterial cell wall synthesis. In mycobacteria, cell elongation occurs through insertion of nascent cell wall material in the sub-polar region, a process largely driven by High Molecular Weight PBPs. In contrast, the function of DD-carboxypeptidases (DD-CPases), which are Low Molecular Weight Class 1C PBPs, in mycobacteria remains poorly understood. Mycobacterium smegmatis encodes four putative DD-CPase homologues, which display homology to counterparts in Escherichia coli. Herein, we demonstrate that these are expressed in varying abundance during growth. Deletion of MSMEG_1661, MSMEG_2433 or MSMEG_2432, individually resulted in no defects in growth, cell morphology, drug susceptibility or spatial incorporation of new peptidoglycan. In contrast, deletion of MSMEG_6113 (dacB) was only possible in a merodiploid strain expressing the homologous M. tuberculosis operon encoding Rv3627c (dacB), Rv3626c, Rv3625c (mesJ) and Rv3624c (hpt), suggestive of essentiality. To investigate the role of this operon in mycobacterial growth, we depleted gene expression using anhydrotetracycline-responsive repressors and noted reduced bipolar peptidoglycan synthesis. These data point to a possible role for this four gene operon, which is highly conserved across all mycobacterial species, in regulating spatial localization of peptidoglycan synthesis.

© 2015 International Society for Microbial Ecology. Prochlorococcus is the numerically dominant phototroph in the oligotrophic subtropical ocean and carries out a significant fraction of marine primary productivity. Although field studies have provided evidence for nitrate uptake by Prochlorococcus, little is known about this trait because axenic cultures capable of growth on nitrate have not been available. Additionally, all previously sequenced genomes lacked the genes necessary for nitrate assimilation. Here we introduce three Prochlorococcus strains capable of growth on nitrate and analyze their physiology and genome architecture. We show that the growth of high-light (HL) adapted strains on nitrate is ∼17% slower than their growth on ammonium. By analyzing 41 Prochlorococcus genomes, we find that genes for nitrate assimilation have been gained multiple times during the evolution of this group, and can be found in at least three lineages. In low-light adapted strains, nitrate assimilation genes are located in the same genomic context as in marine Synechococcus. These genes are located elsewhere in HL adapted strains and may often exist as a stable genetic acquisition as suggested by the striking degree of similarity in the order, phylogeny and location of these genes in one HL adapted strain and a consensus assembly of environmental Prochlorococcus metagenome sequences. In another HL adapted strain, nitrate utilization genes may have been independently acquired as indicated by adjacent phage mobility elements; these genes are also duplicated with each copy detected in separate genomic islands. These results provide direct evidence for nitrate utilization by Prochlorococcus and illuminate the complex evolutionary history of this trait.

The mammalian ovary is composed of ovarian follicles, each follicle consisting of a single oocyte surrounded by somatic granulosa cells, enclosed together within a basement membrane. A finite pool of follicles is laid down during embryonic development, when oocytes in meiotic arrest form a close association with flattened granulosa cells, forming primordial follicles. By or shortly after birth, mammalian ovaries contain their lifetime’s supply of primordial follicles, from which point onwards there is a steady release of follicles into the growing follicular pool.The ovary is particularly amenable to development in vitro, with follicles growing in a highly physiological manner in culture. This work describes the culture of whole neonatal ovaries containing primordial follicles, and the culture of individual ovarian follicles, a method which can support the development of follicles from an immature through to the preovulatory stage, after which their oocytes are able to undergo fertilization in vitro. The work outlined here uses culture systems to determine how the ovary is affected by exposure to external compounds. We also describe a co-culture system, which allows investigation of the interactions that occur between growing follicles and the non-growing pool of primordial follicles.

One of the most widely studied organisms responsible for Harmful Algal Blooms (HABs) is the marine dinoflagellate Karenia brevis. This organism produces neurotoxic compounds known as brevetoxins. A related dinoflagellate, Karenia papilionacea, has been reported to occasionally co-bloom with K. brevis but has received little attention as a possible toxin producing species. Therefore, our aim was to investigate the toxin profile for K. papilionacea. A toxic fraction was identified using a cell based cytotoxicity assay and the toxin was isolated and identified as the ladder frame polyether brevetoxin-2 (PbTx-2) using mass spectrometry (MS) and nuclear magnetic resonance (NMR). Toxin production in K. papilionacea increased in response to hypoosmotic stress, as previously observed in K. brevis.

An updated inventory of Brazilian seed plants is presented and offers important insights into the country's biodiversity. This work started in 2010, with the publication of the Plants and Fungi Catalogue, and has been updated since by more than 430 specialists working online. Brazil is home to 32,086 native Angiosperms and 23 native Gymnosperms, showing an increase of 3% in its species richness in relation to 2010. The Amazon Rainforest is the richest Brazilian biome for Gymnosperms, while the Atlantic Rainforest is the richest one for Angiosperms. There was a considerable increment in the number of species and endemism rates for biomes, except for the Amazon that showed a decrease of 2.5% of recorded endemics. However, well over half of Brazillian seed plant species (57.4%) is endemic to this territory. The proportion of life-forms varies among different biomes: trees are more expressive in the Amazon and Atlantic Rainforest biomes while herbs predominate in the Pampa, and lianas are more expressive in the Amazon, Atlantic Rainforest, and Pantanal. This compilation serves not only to quantify Brazilian biodiversity, but also to highlight areas where there information is lacking and to provide a framework for the challenge faced in conserving Brazil's unique and diverse flora. Resumo Um levantamento atualizado das plantas com sementes e análises relevantes acerca desta biodiversidade são apresentados. Este trabalho se iniciou em 2010 com a publicação do Catálogo de Plantas e Fungos e, desde então vem sendo atualizado por mais de 430 especialistas trabalhando online. O Brasil abriga atualmente 32.086 espécies nativas de Angiospermas e 23 espécies nativas de Gimnospermas e estes novos dados mostram um aumento de 3% da riqueza em relação a 2010. A Amazônia é o Domínio Fitogeográfico com o maior número de espécies de Gimnospermas, enquanto que a Floresta Atlântica possui a maior riqueza de Angiospermas. Houve um crescimento considerável no número de espécies e nas taxas de endemismo para a maioria dos Domínios (Caatinga, Cerrado, Floresta Atlântica, Pampa e Pantanal), com exceção da Amazônia que apresentou uma diminuição de 2,5% de endemicidade. Entretanto, a maior parte das plantas com sementes que ocorrem no Brasil (57,4%) é endêmica deste território. A proporção de formas de vida varia de acordo com os diferentes Domínios: árvores são mais expressivas na Amazônia e Floresta Atlântica do que nos outros biomas, ervas são dominantes no Pampa e as lianas apresentam riqueza expressiva na Amazônia, Floresta Atlântica e Pantanal. Este trabalho não só quantifica a biodiversidade brasileira, mas também indica as lacunas de conhecimento e o desafio a ser enfrentado para a conservação desta flora.