Remote, subsurface detection of the algal toxin domoic acid onboard the Environmental Sample Processor: Assay development and field trials

The ability to detect harmful algal bloom (HAB) species and their toxins in real- or near real-time is a critical need for researchers studying HAB/toxin dynamics, as well as for coastal resource managers charged with monitoring bloom populations in order to mitigate their wide ranging impacts. The Environmental Sample Processor (ESP), a robotic electromechanical/fluidic system, was developed for the autonomous, subsurface application of molecular diagnostic tests and has successfully detected several HAB species using DNA probe arrays during field deployments. Since toxin production and thus the potential for public health and ecosystem effects varies considerably in natural phytoplankton populations, the concurrent detection of HAB species and their toxins onboard the ESP is essential. We describe herein the development of methods for extracting the algal toxin domoic acid (DA) from Pseudonitzschia cells (extraction efficiency >90%) and testing of samples using a competitive ELISA onboard the ESP. The assay detection limit is in the low ng/mL range (in extract), which corresponds to low ng/L levels of DA in seawater for a 0.5 L sample volume acquired by the ESP. We also report the first in situ detection of both a HAB organism (i.e., Pseudo-nitzschia) and its toxin, domoic acid, via the sequential (within 2-3 h) conduct of species- and toxin-specific assays during ESP deployments in Monterey Bay, CA, USA. Efforts are now underway to further refine the assay and conduct additional calibration exercises with the aim of obtaining more reliable, accurate estimates of bloom toxicity and thus their potential impacts. Published by Elsevier B.V.

Development of a planar waveguide microarray for the monitoring and early detection of five harmful algal toxins in water and cultures

A novel multiplex microarray has been developed for the detection of five groups of harmful algal and cyanobacterial toxins found in marine, brackish, and freshwater environments including domoic acid (DA), okadaic acid (OA, and analogues), saxitoxin (STX, and analogues), cylindrospermopsin (CYN) and microcystins (MC, and analogues). The sensitivity and specificity were determined and feasibility to be used as a screening tool investigated. Results for algal/cyanobacterial cultures (n = 12) and seawater samples (n = 33) were compared to conventional analytical methods, such as high performance liquid chromatography (HPLC) and liquid chromatography tandem mass spectrometry (LC-MS/MS). Detection limits for the 15 min assay were 0.37, 0.44, 0.05, 0.08, and 0.40 ng/mL for DA, OA, STX, CYN, and MC, respectively. The correlation of data obtained from the microarray compared to conventional analysis for the 12 cultures was r(2) = 0.83. Analysis of seawater samples showed that 82, 82, 70, 82, and 12% of samples were positive (>IC20) compared to 67, 55, 36, 0, and 0% for DA, OA, STX, CYN, and MC, respectively, for conventional analytical methods. The discrepancies in results can be attributed to the enhanced sensitivity and cross-reactivity profiles of the antibodies in the MBio microarray. The feasibility of the microarray as a rapid, easy to use, and highly sensitive screening tool has been illustrated for the five-plex detection of biotoxins. The research demonstrates an early warning screening assay to support national monitoring agencies by providing a faster and more accurate means of identifying and quantifying harmful toxins in water samples.

Interpreting submarine structures for glacial retreat periodicity within Muchalat Inlet in Nootka Sound

Senior thesis written for Oceanography 445

Characterizing the controls on surface alkalinity in Nootka Sound, British Columbia

Senior thesis written for Oceanography 445

A geochemical and paleoceanographic investigation using marine molluscs of the late Quaternary marine submergences Quebec, Ontario, British Columbia

Sedjrrlents deposited in the Late Quaternary marine sUbrnergences that follov'ted the deglaciation of Ontario} Quebec., and 6ritlst-1 Columbia often contaln an abundant nlarlne invertebrate macrofauna. The rnacrofauna~ dotYllnated by aragonitic pelecypods} is fully preserved In their original mineralogy and cherrlistry 8S deternl1ned by x-ray dlffractlon., scannlng electron tl-,lcroscoDY., trace and r1l1 nor elet11ent analyses and stable isotopes. Ttle trace elernent and stable isotope geochen-Ilstry of chernlcal1y unaltered aragorlitlc molluscs can be used to determine paleoter1-lperatures and paleosallnltles." HO\Never} corrections need to be tllade \fvtlen deterrTIlnlng oxygen-isotope paleotenlperi:ttures due to the lnfluence of isotopically 11gtlt glaciol rneltv-laters and reduced sal1nltles. Ttle eastern Laurentide Ice Sheet probably had an o:~ygen lS0tOP1C composition as low as -8e) 0/00 (Sr1[IW). In additl0fl} corrections need to be rnade to the carbonlsotope values, before salinity deterrnlnatlons are t11ade., due to the reJjuctlon of the terrestrial carbon bl0rnass during glac1al maxlrna. Using geochernlcal data frot11 537 marlne n-'8crolnvertebrates frorTI 72 localities in soutt-,easter Ontarl0 and southern Quebec, it tras been deterrnined that the Late Quaternary Char1lplaln Sea \N6S density stratified along salinity and temperatlJre gradients. The deep-\h/aters of tt-,e Charnplaln Sea tlad salinities that ranged frorn 31 to 36 ppt} and terrlperatures of 00 to 5°C. Conversely.. the st1alloy./-\f*later regirrle of ttle Ctlarnplaln Sea tlad sal1nltles that ranged fron-, 24 to 33 ppt} Y.tltt1 terrlperatures ranglng from 5° to 15°C. Tr,8 rrlajorl rnlnor1 and trace e1et1-,ent geochernlcal analysls of 155 marine lnvertebrates frorn 4 10C611t1es of tt-,e Late Quaternary Ft. Langley Forrnatlon and Capl1ano Sedlments;. souttl\Nestern Brltlsh Columblal suggest l t~lat the 'waters of the o-,arlne lnundation that fol1o....ved the retreating Cordl11eran Ice Sheet had sal1nltles ranglng frorn 32 to 3f. DPt.