Anatomy of a red tide bloom off the southwest coast of Florida

A massive outbreak of Karenia brevis that had been ongoing for several months along the southwestern coast of Florida was sampled in early September 2005 off Sanibel Island to assess the utility of bio-optical features and ataxonomic analysis (quantification of eukaryotic and cyanobacterial picoplankton) by flow cytometry in monitoring red tide blooms. Sea-surface sampling followed aircraft visual location of discolored water. Within the most concentrated area of the bloom, chlorophyll a values exceeded 500 μg l⁻¹, and concentrations of nitrate (0.3 μM ± 0.0) and ammonium (<0.2 μM) were depleted compared to high concentrations of total dissolved nitrogen, total dissolved phosphorus, and soluble reactive phosphorus (141 ± 34 μM, 16.5 ± 2.5 μM, and 6.44 ± 0.57 μM, respectively). Low water clarity in the bloom (Secchi depth transparency 0.3 m, K_d estimated at 4.83 m⁻¹) was strongly influenced by attenuation from dinoflagellates as well as chromophoric dissolved organic matter (CDOM). The fact that the K. brevis bloom occurred in lower-salinity (30 psu), high-nutrient waters implicates riverine transport of land-based nutrients as a source of nutrient supplies that fueled or sustained the bloom. Throughout ongoing efforts to advance modeling and technological capabilities that presently lack reliable predictive capability, bio-optical remote sensing via aerial flyovers along with in-water sensor data can continue to provide accurate coverage of relatively large temporal and spatial features. Flow cytometry can provide conservative (because of some cell lysis), rapid, near-real-time validation of bloom components. The concentration and position of the organisms, along with water mass scalars, can also help to diagnose factors promoting K. brevis bloom development and dispersion.     

Chimeric plastid proteome in the Florida "red tide" dinoflagellate Karenia brevis

Current understanding of the plastid proteome comes almost exclusively from studies of plants and red algae. The proteome in these taxa has a relatively simple origin via integration of proteins from a single cyanobacterial primary endosymbiont and the host. However, the most successful algae in marine environments are the chlorophyll c-containing chromalveolates such as diatoms and dinoflagellates that contain a plastid of red algal origin derived via secondary or tertiary endosymbiosis. Virtually nothing is known about the plastid proteome in these taxa. We analyzed expressed sequence tag data from the toxic "Florida red tide" dinoflagellate Karenia brevis that has undergone a tertiary plastid endosymbiosis. Comparative analyses identified 30 nuclear-encoded plastid-targeted proteins in this chromalveolate that originated via endosymbiotic or horizontal gene transfer (HGT) from multiple different sources. We identify a fundamental divide between plant/red algal and chromalveolate plastid proteomes that reflects a history of mixotrophy in the latter group resulting in a highly chimeric proteome. Loss of phagocytosis in the "red" and "green" clades effectively froze their proteomes, whereas chromalveolate lineages retain the ability to engulf prey allowing them to continually recruit new, potentially adaptive genes through subsequent endosymbioses and HGT. One of these genes is an electron transfer protein (plastocyanin) of green algal origin in K. brevis that likely allows this species to thrive under conditions of iron depletion.     

An epizootic of fibromatosis in gray squirrels (Sciurus carolinensis) in Florida

Beginning in the fall of 1998 and extending into the spring and early summer of 1999 there was a large epizootic of squirrel fibromatosis in squirrels in seven counties in peninsular Florida. Hundreds of gray squirrels (Sciurus carolinensis) with multiple cutaneous tumors were submitted or reported to biologists, veterinary hospitals, and private wildlife rehabilitators. Most squirrels died or were euthanized soon after submission. Twenty squirrels were submitted for necropsy. The majority of the squirrels examined were adults (12/20) and male (15/20). The number and location of tumors varied widely among the affected squirrels; however, a consistent finding was involvement of the eyelids (20/20). Histopathology revealed a proliferative population of mesenchymal cells within the dermis and marked ballooning degeneration of keratinocytes in the overlying epidermis. Intracytoplasmic viral inclusions were present in the neoplastic mesenchymal cell population and the degenerating keratinocytes. Ulceration and necrosis of the surface of the tumors or associated tissues was present in 14 of the 20 squirrels. Virions consistent with poxvirus were observed via electron microscopy in samples collected from a representative tumor. Death of the squirrels was attributed to emaciation, tissue damage, and severe negative energy balance associated with poxvirus infection and massive tumor growth. The underlying cause of this unusual epizootic of fibromatosis in gray squirrels remains unknown.     

Why no red tide was observed on the West Florida Continental Shelf in 2010

Over the past two decades, the two most anomalous years for water properties on the west Florida continental shelf were 1998 and 2010. In both instances, the shelf was ventilated by relatively cold, nutrient-rich waters of deep ocean origin, which reset the background state underlying shelf ecology. The ventilation in both of these years derived from prolonged interactions of the Gulf of Mexico Loop Current with the shelf slope near the Dry Tortugas located on the southwest corner of the shelf. By contacting relatively shallow isobaths, the boundary current interactions there set the entire shelf into motion, facilitating upwelling across the shelf break, even to DeSoto Canyon some 500 km away, and then across the shelf to the near shore. Such prolonged and intense upwelling of nutrient-rich water in 2010 contrasted with the more typically occurring locally wind driven upwelling conditions, whereby waters upwelled at the near shore are from the inner shelf, versus the deep ocean. Thus not all upwelling scenarios have similar consequences. Whereas the typical wind driven upwelling scenario is necessary for Karenia brevis red tide blooms to manifest along the coastline, the rarer, deep ocean induced upwelling scenario (as occurred in 1998 and particularly in 2010) acts to suppress K. brevis red tides because of the elevated inorganic nutrient conditions that they facilitate. Hence, minimal cell counts above background were observed in 1998, and no cell counts above background were observed in 2010. We conclude that the lack of red tide along the west coast of Florida in 2010 was due to anomalously large and protracted upwelling of nutrient-rich waters of deep ocean origin caused by Loop Current and eddy interactions with the shelf slope.     

Uptake of iron by Gomphosphaeria aponina, a possible control organism for the Florida red tide Pytochodiscus brevis.

The assimilation of iron, a growth-limiting metal ion of the cytotoxic marine cyanobacterium, Gomphosphaeria aponina, has been examined in both static and steady-state cultures using 59Fe (III). Uptake of iron by cells followed first-order kinetics, and biphasic (absorption and uptake) behavior was observed as suggested by noted differences between cultures incubated in the light and in the dark. Iron removal in illuminated cultures was rapid, occurring at rates comparable to exponential growth rates. Although uptake was mediated by a chelating agent (EDTA), synthesis and iron assisted transport by hydroxamate-type siderophores was not involved in the uptake of iron by cells, as determined by standard chemical and biological assays of iron deficient cultures. The ecological implications of this research is considered with respect to the cytotoxic antagonism between the cyanobacterium and Florida's red tide organism, Pytochodiscus brevis (Gymnodinium breve).     

Mixotrophy in red tide algae raphidophytes

Marine raphidophytes are common red tide organisms that are distributed worldwide. They are known to be harmful to other plankton and fish and have often caused large-scale fish mortality in many countries. Thus, the population dynamics of raphidophytes is a critical concern for scientists, the aquaculture industry, and government officers from many countries. Raphidophyte growth and mortality should be investigated to understand bloom dynamics. Raphidophytes were thought to be exclusively autotrophic organisms. However, several recent studies have revealed that raphidophytes are able to feed on heterotrophic and autotrophic bacteria, i.e. raphidophytes are mixotrophic algae. Further, high-resolution video microscopy has revealed the mechanism by which raphidophytes feed on bacteria, which involves capturing prey cells in the mucus excreted by mucocysts and engulfing the cells through mucocysts. These discoveries may influence the conventional view on both raphidophyte bloom dynamics and plankton energy flow and carbon cycling. In the present study, I review prey, feeding mechanisms, and ingestion rates of mixotrophic marine raphidophytes. In addition, I examine the ecological significance of raphidophyte mixotrophy.     

Isolation and properties of isolated nuclei from the Florida red tide dinoflagellate Gymnodinium breve (Davis)

A simple and rapid method is described for the isolation of nuclei from the Florida red tide dinoflagellate Gymnodinium breve. The nuclei are free of cytoplasmic contamination and are active in endogenous RNA synthesis. The ratio of DNA : RNA : acid-soluble protein : acid-insoluble protein is 1:0.39:0.13:0.63, respectively, and each nucleus contains ca. 113 picograms of DNA. Electrophoretic analysis of the acid-soluble proteins reveals the presence of two histone-like proteins with molecular weights of 12,000 and 13,000.     

Inorganic carbon acquisition in red tide dinoflagellates

Carbon acquisition was investigated in three marine bloom-forming dinollagellates-Prorocentrum minimum, Heterocapsa triquetra and Ceratium lineatum. In vivo activities of extracellular and intracellular carbonic anhydrase (CA), photosynthetic O2 evolution, CO2 and HCO3- uptake rates were measured by membrane inlet mass spectrometry (MIMS) in cells acclimated to low pH (8.0) and high pH (8.5 or 9.1). A second approach used short-term 14C-disequilibrium incubations to estimate the carbon source utilized by the cells. All three species showed negligible extracellular CA (eCA) activity in cells acclimated to low pH and only slightly higher activity when acclimated to high pH. Intracellular CA (iCA) activity was present in all three species, but it increased only in P. minimum with increasing pH. Half-saturation concentrations (K1/2) for photosynthetic O2 evolution were low compared to ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) kinetics. Moreover, apparent affinities for inorganic carbon (Ci) increased with increasing pH in the acclimation, indicating the operation of an efficient CO2 concentration mechanism (CCM) in these dinoflagellates. Rates of CO2 uptake were comparably low and could not support the observed rates of photosynthesis. Consequently, rates of HCO3- uptake were high in the investigated species, contributing more than 80% of the photosynthetic carbon fixation. The affinity for HCO3- and maximum uptake rates increased under higher pH. The strong preference for HCO3- was also confirmed by the 14C-disequilibrium technique. Modes of carbon acquisition were consistent with the 13C-fractionation pattern observed and indicated a strong species-specific difference in leakage. These results suggest that photosynthesis in marine dinoflagellates is not limited by Ci even at high pH, which may occur during red tides in coastal waters.     

An episode of accelerated amino acid change in Drosophila esterase-6 associated with a change in physiological function

In most lineages of the subgenus Sophophora esterase-6 is a homodimeric haemolymph protein. In the melanogaster subgroup of species it has become a monomer which is mainly expressed in the male sperm ejaculatory duct. Our analyses of esterase-6 sequences from three melanogaster subgroup species and two close relatives reveal a brief period of accelerated amino acid sequence change during the transition between the ancestral and derived states. In this period of 2–6Myr the ratio of replacement to silent site substitutions (0.51) is about three times higher than the values in other lineages of the phylogeny. There are about 50 more replacements in this period than would be predicted from the ratios of replacement to silent site substitutions found elsewhere in the phylogeny. Modelling on the known structure of a related acetylcholinesterase suggests that an unusually high proportion of the replacements in the transitional branch are non-conservative changes on the protein surface. Up to half the accelerated replacement rate can be accounted for by clusters of changes to the face of the molecule containing the opening of the active site gorge. This includes changes in and around regions homologous to peripheral substrate binding sites in acetylcholinesterase. There are also three changes in glycosylation status. One region predicted to lie on the protein surface which becomes markedly more hydrophilic is proposed to be the ancestral dimerisation site that is lost in the transitional branch. This revised version was published online in July 2006 with corrections to the Cover Date.     

Molecular characterization of epizootic hemorrhagic disease virus serotype 1 associated with a 1999 epizootic in white-tailed deer in the eastern United States

During the autumn of 1999 (mid-August-late September), an outbreak of hemorrhagic disease in white-tailed deer (Odocoileus virginianus) caused by epizootic hemorrhagic disease virus serotype 1 (EHDV-1) occurred along the east coast of the United States from Georgia to New Jersey. An EHDV-1 epizootic of such magnitude had not been described in this region since 1975. To determine the genetic relatedness among the 1999 viruses, as well as among additional EHDV-1 isolates from the eastern and western United States, portions of the S10 and L2 gene segments were sequenced and compared utilizing phylogenetic analyses. Nearly all of the 1999 eastern isolates were identical in nucleotide sequence at one or both loci. Additionally, confirmed cases of EHDV-1 in white-tailed deer occurred in a south (Georgia)-to-north (New Jersey/Virginia) progression over a short period of approximately six weeks. Taken together, these results indicate that this outbreak resulted from the spread of a single viral strain. The phylograms derived from analysis of the entire sample set displayed eastern and western region-specific clusterings (topotypes), as well as an eastern versus western difference in branch lengths, which may reflect the influence of epizootic versus enzootic transmission patterns on viral genetic diversity.     

Identification of a poison in toxic scallops from a Gonyaulax tamarensis red tide

The poison in stored extracts of the hepatopancreas of scallops that became toxic during a red tide caused by $\text{Gonyaulax tamarensis}$ has been extensively purified and the product shown to be identical to saxitoxin in its toxicity to mice and TLC behavior in six solvent systems.     

An epizootic Staphylococcus infection in a nude mouse colony

An epizootic of Staphylococcus infection causing abscesses was encountered in a small-scale breeding colony of nude mice of BALB/c background. The incidence of abscess was sporadic and mostly nude (nu/nu) mice aged over 3 months were affected. Staphylococcus aureus was isolated from the face, oral cavity, and feces of almost all nu/nu and heterozygous (nu/+) mice in the colony. After a prolonged time period, up to 10 to 14 months of age, almost all the S. aureus-carrying nu/nu mice produced abscesses and eventually died. Athymicity of the host seemed to be a prerequisite for the abscess formation since nu/+ mice were spared from the lesions. Also, transfer of immunocompetent spleen cells cured the abscesses of the affected nude mice.     

An encephalomyocarditis virus epizootic in a baboon colony.

Approximately 80 baboon deaths were caused by encephalomyocarditis virus (EMCV) infection in a 3060 member research and production colony. The epizootic extended over a 9-month period and occurred in baboons ranging from 1 day to 22 years of age. Acute death was the most common history. When clinical disease was detected, it was characterized by labored respiration associated with acute congestive heart failure. The salient necropsy findings were pulmonary congestion and edema, hydropericardium, hydrothorax, ascites, lymph node and splenic hypertrophy, and pale white-to-tan mottled hearts. The most significant histologic lesion was nonsuppurative necrotizing myocarditis. Placental infection with fetal loss occurred. Diagnosis was confirmed by light microscopy, transmission electron microscopy, virus culture, and serology. Rarely, EMCV-induced antibody persisted in surviving baboons for more than 24 months. EMCV-infected feral rats were the probable source of the virus and their control stopped the epizootic. No EMCV neutralizing antibody was detected in colony support personnel or chimpanzees.     

An epizootic form of Tyzzer's disease in a rat colony

An epizootic form of Tyzzer's disease was experienced in a rearing facility of laboratory rats with an infectivity and morbidity of 36 to 80% and 5 to 30%, respectively. With liver tissue from an affected rat, a spore-forming causative agent was successfully transmitted to other rats by oral inoculation in association with cortisone, producing 10(6) to 10(7) spores/g liver tissue.