Oceanic Microplankton Do Not Adhere to the Latitudinal Diversity Gradient.

A latitudinal biodiversity gradient has captivated ecologists for years, and has become a widely recognized pattern in biogeography, manifest as an increase in biodiversity from the poles to the tropics. Oceanographers have attempted to discern whether these distribution patterns are shared with marine biota, and a lively debate has emerged concerning the global distribution of microbes. Limitations in sampling resolution for such large-scale assessments have often prohibited definitive conclusions. We evaluated microbial planktonic communities along a ~ 15,400-km Pacific Ocean transect with DNA from samples acquired every 2 degrees of latitude within a 3-month period between late August and early November 2003. Next-generation sequencing targeting the Bacteria, Archaea, and Eukarya yielded ~ 10.8 million high-quality sequences. Beta-analysis revealed geographic patterns of microbial communities, primarily the Bacteria and Archaea domains. None of the domains exhibited a unimodal pattern of alpha-diversity with respect to latitude. Bacteria communities increased in richness from Arctic to Antarctic waters, whereas Archaea and Eukarya communities showed no latitudinal or polar trends. Based on our analyses, environmental factors related to latitude thought to influence various macrofauna may not define microplankton diversity patterns of richness in the global ocean.

Surveillance of Microsporidia and Protozoan Pathogens in Pensacola Florida: A One-year Study.

Pathogens and the potential risk they present to public health in recreational waters are of continual public concern. The focus of this study was a year-long sampling campaign to document the presence of Microsporidia and protozoan pathogens in the Bayou Texar waterway in Pensacola, Florida. We used biofilms as sentinel indicators for trapping pathogens in five different locations in Pensacola, Florida. Of the 34 biofilm samples, 16 were positive for pathogens. Of these samples, 13 were positive for Enchephalitozoon spp. (mostly E. cuniculi), 11 were positive for Enterocytozoon bieneusi, and two were positive for Cryptosporidium parvum. The data demonstrate that Microsporidia were easily recovered and primarily present in water during summer months.

Biofilms for Monitoring Presence of Microsporidia in Environmental Water.

The development of molecular methodologies for targeting pathogens such as the Microsporidia has greatly improved our monitoring capabilities and initiatives. This study analyzed samples collected from five locations in Pensacola, Florida, USA for the presence of Microsporidian pathogens. To circumvent various impediments associated with water collection and filtration, we utilized biofilms as sentinels for detection of Microsporidia. We implemented membrane-dissolution and sample purification in a single confined step followed by real-time PCR to confirm pathogen presence. The results of this study demonstrate that microsporidia are present in environmental water sites in the Florida panhandle and that biofilms may serve as another alternative mode to circumvent filtration methods for their detection.

Fine-scale population structure of the northern hard clam (Mercenaria mercenaria) revealed by genome-wide SNP markers.

Aquaculture is growing rapidly worldwide, and sustainability is dependent on an understanding of current genetic variation and levels of connectivity among populations. Genetic data are essential to mitigate the genetic and ecological impacts of aquaculture on wild populations and guard against unintended human-induced loss of intraspecific diversity in aquacultured lines. Impacts of disregarding genetics can include loss of diversity within and between populations and disruption of local adaptation patterns, which can lead to a decrease in fitness. The northern hard clam, Mercenaria mercenaria (Linnaeus, 1758), is an economically valuable aquaculture species along the North American Atlantic and Gulf coasts. Hard clams have a pelagic larval phase that allows for dispersal, but the level of genetic connectivity among geographic areas is not well understood. To better inform the establishment of site-appropriate aquaculture brood stocks, this study used DArTseq™ genotyping by sequencing to characterize the genetic stock structure of wild clams sampled along the east coast of North America and document genetic diversity within populations. Samples were collected from 15 locations from Prince Edward Island, Canada, to South Carolina, USA. Stringent data filtering resulted in 4960 single nucleotide polymorphisms from 448 individuals. Five genetic breaks separating six genetically distinct populations were identified: Canada, Maine, Massachusetts, Mid-Atlantic, Chesapeake Bay, and the Carolinas (F ST 0.003-0.046; p < 0.0001). This is the first study to assess population genetic structure of this economically important hard clam along a large portion of its native range with high-resolution genomic markers, enabling identification of previously unrecognized population structure. Results of this study not only broaden insight into the factors shaping the current distribution of M. mercenaria but also reveal the genetic population dynamics of a species with a long pelagic larval dispersal period along the North American Atlantic and Gulf coasts.

Accumulation of organic and inorganic contaminants in shellfish collected in estuarine waters near Pensacola, Florida: contamination profiles and risks to human consumers.

We conducted a screening level assessment of contaminants in blue crabs (Callinectes sapidus) and oysters (Crassostrea virginica) from bays and bayous in the Pensacola, FL area. Tissue samples were analyzed for 17 dioxins/furans, 12 dioxin-like PCB (DL-PCBs) congeners, mercury, and various metals. Contaminant levels were compared to screening values (SV) calculated using U.S. EPA recommendations for establishing consumption advisories. All sampling locations exceeded the SV (0.098pgg(-1)) for dioxins/furans/DL-PCBs, based on a Florida-specific consumption rate (46gday(-1)). Arsenic (inorganic), mercury, cadmium, and zinc levels exceeded SVs in samples from select locations, and with the exception of mercury, these locations were generally downstream of known contaminated areas. We also assessed potential human health risks from consumption of these species. Risks to human health were greatest from consumption of crab hepatopancreas, suggesting that consumption of hepatopancreas, whether directly or indirectly, from crabs collected anywhere in the Pensacola Bay region should be avoided.

Effective concentration and detection of cryptosporidium, giardia, and the microsporidia from environmental matrices.

Cryptosporidium spp., Giardia spp., and members of Microsporidia are enteropathogenic parasites of humans and animals, producing asymptomatic to severe intestinal infections. To circumvent various impediments associated with current detection methods, we tested a method providing multistage purification and separation in a single, confined step. Standard real-time PCR was used as a detection method. Samples spiked with C. parvum and G. intestinalis were split for comparison to standard Method 1623. Results were equivalent to immunomagnetic procedures for Cryptosporidium, and Giardia. Overall percent recovery for Cryptosporidium with Method 1623 averaged 26.89% (std 21.44%; min = 0%; max = 73%) and was similar but less variable for qPCR method at an estimated average of 27.67 (std 17.65%; min = 5%; max = 63%). For Giardia, Method 1623 had an overall average recovery of 27.11% (std 17.98%; min = 1%; max = 58%), while multistage purification and qPCR had an estimated lower overall recovery at 18.58% (std 13.95%; min = 0%; max = 35%). Microsporidia were also readily detected with an estimated recovery of 46.81% overall (std 17.66%; min = 18%; max = 70%) for E. intestinalis and 38.90% (std 14.36%; min = 13%; max = 62%) for E. bieneusi.

PAH concentrations in Coquina (Donax spp.) on a sandy beach shoreline impacted by a marine oil spill.

The BP MC252 well failure in the Gulf of Mexico, April 2010 caused concern for crude oil and polycyclic aromatic hydrocarbon (PAHs) exposure along the sandy beaches of the Florida Panhandle. We began collections of Coquina clams (Donax spp.) from the surf zone of Florida Panhandle beaches to monitor PAH contamination to compliment analysis of surf zone sand samples. These clams had higher levels of PAHs relative to ambient sand, and this allowed us to continue to monitor PAH levels after sand concentrations fell below limits of detection. PAH levels in the Coquina tissues were highly variable, perhaps indicative of the heterogeneous distribution of oil and tar on the beaches and exposure to tar particles. Overall, PAH levels decreased continuously in both sand and Coquina tissues, reaching limits of detection within one and two years respectively after oil landed on Florida Panhandle beaches. Our work suggests these surf zone molluscs may be used to monitor pollutant exposure along high energy sandy beach shorelines.

Polycyclic aromatic hydrocarbon concentrations across the Florida Panhandle continental shelf and slope after the BP MC 252 well failure.

The Florida Panhandle continental shelf environment was exposed to oil from the BP oil well failure in the Gulf of Mexico during 2010. Floating mats of oil were documented by satellite, but the distribution of dissolved components of the oil in this region was unknown. Shipek® grab samples of sediments were taken during repeated cruises between June 2010 and June 2012 to test for selected polycyclic aromatic hydrocarbons (PAHs) as indicators of this contamination. Sediments were collected as composite samples, extracted using standard techniques, and PAHs were quantified by GC/MS-SIM. PAHs in samples from the continental slope in May 2011 were highest near to the failed well site and were reduced in samples taken one year later. PAHs from continental shelf sediments during the spill (June 2010) ranged from 10 to 165 ng g(-1). Subsequent cruises yielded variable and reduced amounts of PAHs across the shelf. The data suggest that PAHs were distributed widely across the shelf, and their subsequent loss to background levels suggests these compounds were of oil spill origin. PAH half-life estimates by regression were 70-122 days for slope and 201 days for shelf stations.

Largemouth bass (Micropterus salmoides) and striped mullet (Mugil cephalus) as vectors of contaminants to human consumers in northwest Florida.

The health benefits of regular consumption of fish and seafood have been espoused for many years. However, fish are also a potential source of environmental contaminants that have well known adverse effects on human health. We investigated the consumption risks for largemouth bass (Micropterus salmoides; n = 104) and striped mullet (Mugil cephalus; n = 170), two commonly harvested and consumed fish species inhabiting fresh and estuarine waters in northwest Florida. Skinless fillets were analyzed for total mercury, inorganic arsenic, polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/F), polychlorinated biphenyls (PCBs), and organochlorine pesticides. Contaminant levels were compared to screening values (SV) calculated using U.S. Environmental Protection Agency (EPA) recommendations for establishing consumption advisories. Largemouth bass were found to contain high levels of total mercury at all sampling locations (0.37-0.89 ug/g) and one location exhibited elevated total PCBs (39.4 ng/g). All of the samples exceeded Florida fish consumption advisory trigger levels for total mercury and one location exceeded the U.S. EPA SV for total PCBs. As a result of the high mercury levels, the non-cancer health risks (hazard index-HI) for bass were above 1 for all locations. Striped mullet from several locations with known point sources contained elevated levels of PCBs (overall range 3.4-59.3 ng/g). However, total mercury levels in mullet were low. Eight of the 16 mullet sampling locations exceeded the U.S. EPA SV for total PCBs and two locations exceeded an HI of 1 due to elevated PCBs. Despite the elevated levels of total PCBs in some samples, only two locations exceeded the acceptable cancer risk range and therefore cancer health risks from consumption of bass and mullet were determined to be low at most sampling locations.

Stability and change in estuarine biofilm bacterial community diversity.

Biofouling communities contribute significantly to aquatic ecosystem productivity and biogeochemical cycling. Our knowledge of the distribution, composition, and activities of these microbially dominated communities is limited compared to other components of estuarine ecosystems. This study investigated the temporal stability and change of the dominant phylogenetic groups of the domain Bacteria in estuarine biofilm communities. Glass slides were deployed monthly over 1 year for 7-day incubations during peak tidal periods in East Sabine Bay, Fla. Community profiling was achieved by using 16S rRNA genes and terminal restriction fragment length polymorphism (T-RFLP) of 16S rRNA genes in combination with ribotyping, cloning, and sequencing to evaluate diversity and to identify dominant microorganisms. Bacterial community profiles from biofilms grown near the benthos showed distinct periods of constancy within winter and summer sampling periods. Similar periods of stability were also seen in T-RFLP patterns from floating biofilms. Alternating dominance of phylogenetic groups between seasons appeared to be associated with seasonal changes in temperature, nutrient availability, and light. The community structure appeared to be stable during these periods despite changes in salinity and in dissolved oxygen.

Influence of an oyster reef on development of the microbial heterotrophic community of an estuarine biofilm.

We characterized microbial biofilm communities developed over two very closely located but distinct benthic habitats in the Pensacola Bay estuary using two complementary cultivation-independent molecular techniques. Biofilms were grown for 7 days on glass slides held in racks 10 to 15 cm over an oyster reef and an adjacent muddy sand bottom. Total biomass and optical densities of dried biofilms showed dramatic differences for oyster reef versus non-oyster reef biofilms. This study assessed whether the observed spatial variation was reflected in the heterotrophic prokaryotic species composition. Genomic biofilm DNA from both locations was isolated and served as a template to amplify 16S rRNA genes with universal eubacterial primers. Fluorescently labeled PCR products were analyzed by terminal restriction fragment length polymorphism, creating a genetic fingerprint of the composition of the microbial communities. Unlabeled PCR products were cloned in order to construct a clone library of 16S rRNA genes. Amplified ribosomal DNA restriction analysis was used to screen and define ribotypes. Partial sequences from unique ribotypes were compared with existing database entries to identify species and to construct phylogenetic trees representative of community structures. A pronounced difference in species richness and evenness was observed at the two sites. The biofilm community structure from the oyster reef setting had greater evenness and species richness than the one from the muddy sand bottom. The vast majority of the bacteria in the oyster reef biofilm were related to members of the gamma- and delta-subdivisions of Proteobacteria, the Cytophaga-Flavobacterium -Bacteroides cluster, and the phyla Planctomyces and Holophaga-Acidobacterium. The same groups were also present in the biofilm harvested at the muddy sand bottom, with the difference that nearly half of the community consisted of representatives of the Planctomyces phylum. Total species richness was estimated to be 417 for the oyster reef and 60 for the muddy sand bottom, with 10.5% of the total unique species identified being shared between habitats. The results suggest dramatic differences in habitat-specific microbial diversity that have implications for overall microbial diversity within estuaries.