An ancient river landscape preserved beneath the East Antarctic Ice Sheet.

The East Antarctic Ice Sheet (EAIS) has its origins ca. 34 million years ago. Since then, the impact of climate change and past fluctuations in the EAIS margin has been reflected in periods of extensive vs. restricted ice cover and the modification of much of the Antarctic landscape. Resolving processes of landscape evolution is therefore critical for establishing ice sheet history, but it is rare to find unmodified landscapes that record past ice conditions. Here, we discover an extensive relic pre-glacial landscape preserved beneath the central EAIS despite millions of years of ice cover. The landscape was formed by rivers prior to ice sheet build-up but later modified by local glaciation before being dissected by outlet glaciers at the margin of a restricted ice sheet. Preservation of the relic surfaces indicates an absence of significant warm-based ice throughout their history, suggesting any transitions between restricted and expanded ice were rapid.

Seasonal variations in the skin epidermal structure and mucosal immune parameters of rainbow trout skin (Oncorhynchus mykiss) at different stages of farming.

The aim of this study was to investigate the seasonal changes in the epidermal structure and the innate immunity parameters of skin mucus in rainbow trout. The skin epidermis and mucus samples were collected over three consecutive seasons including winter, spring and late summer from three different weight groups i.e., 2-20 g (W1), 100-200 g (W2) and 400-600 g (W3) fish. The skin mucosal immunity analysis of rainbow trout showed that the haemagglutination activity increased significantly with increasing fish size from W1 to W3 in all three seasons, while no significant seasonal changes occurred in haemagglutination activity. Moreover, the bactericidal activity against fish pathogens increased significantly with increasing water bacterial load in late summer. The SDS-PAGE analysis of mucus showed a high amount of low molecular weight proteins (<35 kDa) in the late summer that was correlated with the increase in bactericidal activity. Histological analysis of the epidermis structure of rainbow trout skin showed that the density and size of goblet cells and consequently the mucus secretion significantly increased in W3 group in all seasons. In all three weight groups of fish, the density of goblet cells significantly increased from winter to spring and late summer along with increasing water temperature. Moreover, the goblet cell density showed a significant positive relationship with the soluble protein concentration and haemagglutination activity (p < 0.01). The results of this study demonstrated the more active immune role of the skin epidermal cells and mucus in rainbow trout during summer to protect fish against the pathogenic microorganisms. Given its potent bactericidal properties and the lack of haemolytic activity, the rainbow trout mucus might be used as a safe and inexpensive source for developing antimicrobial agents to prevent and treat some bacterial diseases in human and fish.

Clinical and Electrophysiological Characteristics of Ventricular Tachycardias From the Basal Septum in Structural Heart Disease.

OBJECTIVES: This study describes the clinical and electrophysiological characteristics of basal-septal ventricular tachycardias (VTs) in patients with structural heart disease (SHD). BACKGROUND: The basal septum is a common source of VT in patients with SHD. METHODS: Data from 312 consecutive patients with SHD undergoing catheter ablation of ventricular arrhythmias were reviewed. RESULTS: Thirty-three basal-septal VTs in 31 patients (mean age 67.4 ± 14.2 years, mean left ventricular ejection fraction [LVEF] 42% ± 15%) were identified. Patients with VTs with left ventricular basal-septal breakthrough were more likely to have ischemic cardiomyopathy and lower LVEF; patients with right ventricular basal-septal VT were more likely to have sarcoidosis or right ventricular cardiomyopathy of unknown significance, with higher LVEF. Atrioventricular block was present in 45% of patients and intraventricular block including persistent biventricular pacing in 77%. Unipolar scar was larger than bipolar scar (area 18.8% ± 19.4% vs 12.7% ± 14.6%; P < 0.001). VTs with right bundle branch block configuration and S wave in lead V6 with positive V3/V4 polarity consistently indicated left ventricular basal-septal breakthrough. Inferior limb-lead discordance with right bundle branch block configuration and "reverse pattern break in lead V2" were identified in left ventricular basal inferior-septal origin in 3 patients. VT noninducibility was achieved in 55%, and VT recurred in 42% of patients after a single procedure, but VT burden was significantly reduced after ablation (59 episodes before vs 2 episodes after ablation; P = 0.02). CONCLUSIONS: Basal-septal VTs in patients with SHD have a distinct clinical, electrocardiographic, and electrophysiological profile depending on the breakthrough site, accompanied by a deep intramural septal substrate that limits procedural success after catheter ablation.

Epicardial-Endocardial Reentry in Ischemic Cardiomyopathy.

In ischemic cardiomyopathy, endocardial reentry has traditionally been the mechanistic paradigm for understanding ventricular tachycardia (VT). However, recognition is growing that epicardial myocardium is a critical component for VT substrate, even in patients with ischemic cardiomyopathy. In this report, we present a novel case of a three-dimensional VT reentry involving epicardial components and an endocardial exit.

Validation of a QuEChERS method for extraction of estrogens from a complex water matrix and quantitation via high-performance liquid chromatography-mass spectrometry.

The traditional approach to extracting estrogens from water matrices, solid-phase extraction (SPE), presents a number of challenges when applied to complex wastewater matrices. Conversely, the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) clean-up method offers an alternative sample preparation approach that omits sample filtration and overcomes additional challenges associated with SPE. The objective of this study was to implement and validate a scaled QuEChERS method, using a standard addition approach, for extracting estrone (E1), 17ß-estradiol (E2), and estriol (E3) from the estrogenic influent of a recirculating aquaculture system containing American eels (Anguilla rostrata). While traditional QuEChERS protocols do not facilitate considerable sample concentration, a 500-fold concentration factor was implemented for reliable quantitation of parts-per-trillion concentrations of estrogens from an initial sample volume of 20 mL to a final extract volume of 40 µL. Following analysis via high-performance liquid chromatography-mass spectrometry, excellent process efficiencies were observed at spiked concentrations of 10 and 50 ng L-1 for E2 and E1 (101 to 111%; %RSD ≤ 16), and moderate to acceptable process efficiencies were achieved for E3 (75 to 87%; %RSD ≤ 16). Validation of method parameters, including specificity, linearity, accuracy (recovery and process efficiencies), precision (intra-day precision, and inter-day precision), matrix effects, method detection limit, and limit of quantitation, led to reliable quantitation of unknown concentrations of E1, E2, and E3 in the aquaculture influent as low as 52, 20, and 33 ng L-1, respectively. This study provides a validated analytical method for waste systems requiring quantitation of estrogens in their complex wastewater matrices.

Hard rock landforms generate 130 km ice shelf channels through water focusing in basal corrugations.

Satellite imagery reveals flowstripes on Foundation Ice Stream parallel to ice flow, and meandering features on the ice-shelf that cross-cut ice flow and are thought to be formed by water exiting a well-organised subglacial system. Here, ice-penetrating radar data show flow-parallel hard-bed landforms beneath the grounded ice, and channels incised upwards into the ice shelf beneath meandering surface channels. As the ice transitions to flotation, the ice shelf incorporates a corrugation resulting from the landforms. Radar reveals the presence of subglacial water alongside the landforms, indicating a well-organised drainage system in which water exits the ice sheet as a point source, mixes with cavity water and incises upwards into a corrugation peak, accentuating the corrugation downstream. Hard-bedded landforms influence both subglacial hydrology and ice-shelf structure and, as they are known to be widespread on formerly glaciated terrain, their influence on the ice-sheet-shelf transition could be more widespread than thought previously.

Toward the first quantum simulation with quantum speedup.

With quantum computers of significant size now on the horizon, we should understand how to best exploit their initially limited abilities. To this end, we aim to identify a practical problem that is beyond the reach of current classical computers, but that requires the fewest resources for a quantum computer. We consider quantum simulation of spin systems, which could be applied to understand condensed matter phenomena. We synthesize explicit circuits for three leading quantum simulation algorithms, using diverse techniques to tighten error bounds and optimize circuit implementations. Quantum signal processing appears to be preferred among algorithms with rigorous performance guarantees, whereas higher-order product formulas prevail if empirical error estimates suffice. Our circuits are orders of magnitude smaller than those for the simplest classically infeasible instances of factoring and quantum chemistry, bringing practical quantum computation closer to reality.

Analysis of 17ß-estradiol, estriol and estrone in American eel (Anguilla rostrata) tissue samples using liquid chromatography coupled to electrospray differential ion mobility tandem mass spectrometry.

RATIONALE: 17ß-Estradiol (E2), estrone (E1) and estriol (E3) are steroid hormones responsible for the regulation of the female reproductive system. Estradiol is planned to be used to feminize eels in aquaculture in order to improve their size and marketability. The residual levels of these hormones in fish tissue must be monitored to meet the requirements of food regulatory agencies. Few studies have studied these hormones in complex biological matrices such as fish tissue. METHODS: We developed a method to analyze E1, E2 and E3 in fish tissue using liquid chromatography in combination with differential ion mobility spectrometry (DMS) and tandem mass spectrometry (MS/MS). The mass spectrometer was operated in negative polarity selected reaction monitoring (SRM) mode. To test the performance of this method, residual levels of E1, E2 and E3 were measured in the muscle tissue of juvenile eels subjected to feminization treatment with E2. RESULTS: We report that following 17ß-estradiol treatment, E2 is rapidly metabolized from the eel tissue, with a 50% depletion rate per day. Five days post-treatment, E2 returned to the level found in non-treated controls, similar to levels found in wild mature female eels. CONCLUSIONS: The method presented herein allows the quantitative analysis of E1, E2 and E3 in fish tissue samples. Under the experimental conditions, E2 in fish tissue samples returned to physiological levels post hormonal treatment. Copyright © 2017 John Wiley & Sons, Ltd.

Slow Pathway Radiofrequency Ablation Using Magnetic Navigation: A Description of Technique and Retrospective Case Analysis.

BACKGROUND: The Magnetic Navigation System (MNS) catheter was shown to be stable in the presence of significant cardiac wall motion and delivered more effective lesions compared to manual control. This stability could potentially make AV junctional re-entrant tachycardia (AVNRT) ablation safer. The aim of this study is to describe the method of mapping and ablation of AVNRT with MNS and 3-D electro-anatomical mapping system (CARTO, Biosense Webster, Diamond bar, CA, USA) anatomical mapping, with a view to improve the safety of ablation. METHODS: The method of precise mapping and ablation with MNS is described. Consecutive AVNRT cases (n=30) from 2012 January to 2015 November, in which magnetic navigation was used, are analysed. RESULTS: Ablation was successful in 27 (90%) out of 30 patients. In three cases, ablation was abandoned due to the proximity of the three-dimensional His image to the potential ablation site. No complications, including AV nodal injury, occurred. The distance from the nearest His position to successful ablation site in both LAO and RAO projections of CARTO images was 26.4±8.8 and 27±7.7mm respectively. Only in two (9%) patients, ablation needed to be extended superior to the plane of coronary sinus ostium, towards the His bundle region, to achieve slow pathway modification. CONCLUSION: AVNRT ablation with MNS allows for accurate mapping of the AV node and stable ablation at a safe distance, which could help avoid AV nodal injury. We recommend this modality for younger patients with AVNRT.

Erythrocyte heat shock protein responses to chronic (in vivo) and acute (in vitro) temperature challenge in diploid and triploid salmonids.

This research investigated how ploidy level (diploid versus triploid) affects the heat shock protein (HSP) response in erythrocytes under different thermal stress regimes, both in vivo and in vitro, in Atlantic salmon (Salmo salar) and brook charr (Salvelinus fontinalis) in order to address the question of why triploids typically have reduced thermal tolerance. A preliminary study confirmed that identical volumes of diploid and triploid erythrocytes (which equates to a smaller number of larger cells for triploids compared to diploids) did not differ in total protein synthesis rates. After chronic (100d) acclimation of fish to 5, 15 and 25°C, triploid erythrocytes had lower HSP70, HSP90, heat shock factor 1 (HSF1) and ubiquitin (free and total) levels than diploids in both species. Furthermore, Atlantic salmon erythrocytes showed significantly higher protein breakdown (based on conjugated ubiquitin levels) in triploids than diploids after acute heat stress in vitro, but no significant difference was detected between ploidies after acute cold stress. These results indicate that: 1) triploid erythrocytes synthesize more total protein per cell than diploids as a result of increased cell size; 2) triploids have sufficient total HSP levels for survival under low stress conditions; and 3) the lower basal titres of HSPs in triploids may be a handicap when combating acute stress. Taken together, this suggests that triploids are limited in their ability to withstand thermal stress because of a reduced ability to maintain proteostasis under stressful conditions.

Clean subglacial access: prospects for future deep hot-water drilling.

Accessing and sampling subglacial environments deep beneath the Antarctic Ice Sheet presents several challenges to existing drilling technologies. With over half of the ice sheet believed to be resting on a wet bed, drilling down to this environment must conform to international agreements on environmental stewardship and protection, making clean hot-water drilling the most viable option. Such a drill, and its water recovery system, must be capable of accessing significantly greater ice depths than previous hot-water drills, and remain fully operational after connecting with the basal hydrological system. The Subglacial Lake Ellsworth (SLE) project developed a comprehensive plan for deep (greater than 3000 m) subglacial lake research, involving the design and development of a clean deep-ice hot-water drill. However, during fieldwork in December 2012 drilling was halted after a succession of equipment issues culminated in a failure to link with a subsurface cavity and abandonment of the access holes. The lessons learned from this experience are presented here. Combining knowledge gained from these lessons with experience from other hot-water drilling programmes, and recent field testing, we describe the most viable technical options and operational procedures for future clean entry into SLE and other deep subglacial access targets.

Recent advances in understanding Antarctic subglacial lakes and hydrology.

It is now well documented that over 400 subglacial lakes exist across the bed of the Antarctic Ice Sheet. They comprise a variety of sizes and volumes (from the approx. 250 km long Lake Vostok to bodies of water less than 1 km in length), relate to a number of discrete topographic settings (from those contained within valleys to lakes that reside in broad flat terrain) and exhibit a range of dynamic behaviours (from 'active' lakes that periodically outburst some or all of their water to those isolated hydrologically for millions of years). Here we critique recent advances in our understanding of subglacial lakes, in particular since the last inventory in 2012. We show that within 3 years our knowledge of the hydrological processes at the ice-sheet base has advanced considerably. We describe evidence for further 'active' subglacial lakes, based on satellite observation of ice-surface changes, and discuss why detection of many 'active' lakes is not resolved in traditional radio-echo sounding methods. We go on to review evidence for large-scale subglacial water flow in Antarctica, including the discovery of ancient channels developed by former hydrological processes. We end by predicting areas where future discoveries may be possible, including the detection, measurement and significance of groundwater (i.e. water held beneath the ice-bed interface).

The distribution of pace adopted by cyclists during a cross-country mountain bike World Championships.

The purpose of this study was to examine the distribution of pace self-selected by cyclists of varying ability, biological age and sex performing in a mountain bike World Championship event. Data were collected on cyclists performing in the Elite Male (ELITEmale; n = 75), Elite Female (ELITEfemale; n = 50), Under 23 Male (U23male; n = 62), Under 23 Female (U23female; n = 34), Junior Male (JNRmale; n = 71) and Junior Female (JNRfemale; n = 30) categories of the 2009 UCI Cross-Country Mountain Bike World Championships. Split times were recorded for the top, middle and bottom 20% of all finishers of each category. Timing splits were positioned to separate the course into technical and non-technical, uphill, downhill and rolling/flat sections. Compared with bottom performers, top performers in all male categories (ELITEmale, U23male, JNRmale) maintained a more even pace over the event as evidenced by a significantly lower standard deviation and range in average lap speed. Top performers, males, and ELITEmale athletes spent a lower percentage of overall race time on technical uphill sections of the course, compared with middle and bottom placed finishers, females, and JNRmale athletes, respectively. Better male performers adopt a more even distribution of pace throughout cross-country mountain events. Performance of lower placed finishers, females and JNRmale athletes may be improved by enhancing technical uphill cycling ability.

Analysis of a ferric uptake regulator (Fur) knockout mutant in Aeromonas salmonicida subsp. salmonicida.

Aeromonas salmonicida subsp. salmonicida is the etiological agent of furunculosis; a serious infectious disease in aquaculture raised salmonids. Iron acquisition has been shown to be critical for the survival of pathogenic bacteria during the course of infection. Previous work has demonstrated that A. salmonicida expresses iron-repressible IROMP proteins, suggesting the presence of iron acquisition systems that are under the control of a ferric uptake regulator (Fur). In this study, the A. salmonicida fur has been sequenced and a fur deletion strain generated. The A. salmonicida fur gene has an open reading frame of 428 bp, coding for a protein of 143 amino acids, and with high homology to previously described Fur proteins. The Fur protein product had a 94% sequence identity and 96% sequence similarity to the Aeromonas hydrophila Fur protein product. Transcription of the A. salmonicida fur gene was not regulated by the iron status of the bacterium and is not autoregulated, as in Escherichia coli. Proteomic analysis of the A. salmonicida fur mutant, fails to repress iron-regulated outer membrane proteins in the presence of iron. The A. salmonicida fur::KO mutant shows significantly reduced pathogenicity compared to the wild-type parental strain. In addition, the A. salmonicida fur mutant provides an important tool for further investigation of the iron acquisition mechanisms utilized by A. salmonicida.

Glutathione, glutathione S-transferase, and glutathione conjugates, complementary markers of oxidative stress in aquatic biota.

Contaminants are ubiquitous in the environment and their impacts are of increasing concern due to human population expansion and the generation of deleterious effects in aquatic species. Oxidative stress can result from the presence of persistent organic pollutants, metals, pesticides, toxins, pharmaceuticals, and nanomaterials, as well as changes in temperature or oxygen in water, the examined species, with differences in age, sex, or reproductive cycle of an individual. The antioxidant role of glutathione (GSH), accompanied by the formation of its disulfide dimer, GSSG, and metabolites in response to chemical stress, are highlighted in this review along with, to some extent, that of glutathione S-transferase (GST). The available literature concerning the use and analysis of these markers will be discussed, focusing on studies of aquatic organisms. The inclusion of GST within the suite of biomarkers used to assess the effects of xenobiotics is recommended to complement that of lipid peroxidation and mixed function oxygenation. Combining the analysis of GSH, GSSG, and conjugates would be beneficial in pinpointing the role of contaminants within the plethora of causes that could lead to the toxic effects of reactive oxygen species.

Up-regulation of hepatic ABCC2, ABCG2, CYP1A1 and GST in multixenobiotic-resistant killifish (Fundulus heteroclitus) from the Sydney Tar Ponds, Nova Scotia, Canada.

Cellular defence against accumulation of toxic xenobiotics includes metabolism by phase I and II enzymes and export of toxicants and their metabolites via ATP-binding cassette (ABC) transporters. Liver gene expression of representatives of these three protein groups was examined in a population of multixenobiotic-resistant killifish (Fundulus heteroclitus) from the Sydney Tar Ponds, Nova Scotia, Canada. The Tar Ponds are heavily polluted with polycyclic aromatic hydrocarbons, polychlorinated biphenyls and heavy metals. The relationship among ABC transporters ABCB1, ABCB11, ABCC2, ABCG2, phase I enzyme cytochrome P4501A1 (CYP1A1) and phase II enzyme glutathione-S-transferase (GST-mu) was investigated by quantifying hepatic transcript abundance. In Tar Pond killifish, hepatic mRNA expression levels of ABCC2, ABCG2, CYP1A1 and GST-mu were elevated compared to reference sites, suggesting that hydrophobic contaminants undergo phase I and II metabolism and are then excreted into the bile of these fish. Hepatic ABCB1 and ABCB11 mRNA were not up-regulated in Tar Pond fish compared to two reference sites, indicating that these two proteins are not involved in conferring multixenobiotic resistance to Tar Pond killifish. The results suggest instead that liver up-regulation of phase I and II enzymes and complementary ABC transporters ABCC2 and ABCG2 may confer contaminant resistance to Tar Pond fish.

Myxinidin, a novel antimicrobial peptide from the epidermal mucus of hagfish, Myxine glutinosa L.

Fish epidermal mucus contains innate immune components that provide a first line of defense against various infectious pathogens. This study reports the bioassay-guided fractionation and characterization of a novel antimicrobial peptide, myxinidin, from the acidic epidermal mucus extract of hagfish (Myxine glutinosa L.). Edman sequencing and mass spectrometry revealed that myxinidin consists of 12 amino acids and has a molecular mass of 1,327.68 Da. Myxinidin showed activity against a broad range of bacteria and yeast pathogens at minimum bactericidal concentration (MBC) ranging from 1.0 to 10.0 microg/mL. Screened pathogens, Salmonella enterica serovar Typhimurium C610, Escherichia coli D31, Aeromonas salmonicida A449, Yersinia ruckeri 96-4, and Listonella anguillarum 02-11 were found to be highly sensitive to myxinidin at the MBC of 1.0-2.5 microg/mL; Staphylococcus epidermis C621 and yeast (Candida albicans C627) had an MBC of 10.0 microg/mL. The antimicrobial activity of myxinidin was found to be two to 16 times more active than a potent fish-derived antimicrobial peptide, pleurocidin (NRC-17), against most of the screened pathogens. The microbicidal activity of myxinidin was retained in the presence of sodium chloride (NaCl) at concentrations up to 0.3 M and had no hemolytic activity against mammalian red blood cells. These results suggest that myxinidin may have potential applications in fish and human therapeutics.

Changes in Atlantic salmon (Salmo salar) epidermal mucus protein composition profiles following infection with sea lice (Lepeophtheirus salmonis).

The mucus protein profile of Atlantic salmon (Salmo salar) and changes due to infection with sea lice (Lepeophtheirus salmonis) were examined. Two-dimensional gel electrophoresis was performed on salmon skin mucus and comparisons between control and infected fish mucus were made. LC MS/MS identified intracellular proteins, calmodulin, actin, and hemopexin and plasma proteins, such as apolipoproteins, lectin, plasminogen and transferrin. Plasma proteins in the mucus may result from either direct expression by epidermal cells, leakage of plasma or via a secondary circulation system. Therefore, RT-PCR was used to measure mRNA of transferrin and lectin in Atlantic salmon skin. Transferrin expression was observed suggesting direct expression by the epidermis. Lectin expression was not detected suggesting another mechanism of entry into mucus, either leakage from plasma or secondary circulation. The lack of observable albumin on 2D gels, suggests that mucus lectin may arise from the secondary circulation route. Interestingly, ?-actin was a significant component of Atlantic salmon mucus. Cleaved actin and transferrin fragments were observed and positively correlated with sea lice infection suggestive of proteolytic activity. Increased levels of cleaved transferrin during sea lice infection may activate the nitrous oxide response of salmon macrophages, as part of the fish's immune response to sea lice infection.

Comparison of antimicrobial activity in the epidermal mucus extracts of fish.

The mucus layer on the surface of fish consists of several antimicrobial agents that provide a first line of defense against invading pathogens. To date, little is known about the antimicrobial properties of the mucus of Arctic char (Salvelinus alpinus), brook trout (S. fontinalis), koi carp (Cyprinus carpio sub sp. koi), striped bass (Morone saxatilis), haddock (Melanogrammus aeglefinus) and hagfish (Myxine glutinosa). The epidermal mucus samples from these fish were extracted with acidic, organic and aqueous solvents to identify potential antimicrobial agents including basic peptides, secondary metabolites, aqueous and acid soluble compounds. Initial screening of the mucus extracts against a susceptible strain of Salmonella enterica C610, showed a significant variation in antimicrobial activity among the fish species examined. The acidic mucus extracts of brook trout, haddock and hagfish exhibited bactericidal activity. The organic mucus extracts of brook trout, striped bass and koi carp showed bacteriostatic activity. There was no detectable activity in the aqueous mucus extracts. Further investigations of the activity of the acidic mucus extracts of brook trout, haddock and hagfish showed that these fish species had specific activity for fish and human pathogens, demonstrating the role of fish mucus in antimicrobial protection. In comparison to brook trout and haddock, the minimum bactericidal concentrations of hagfish acidic mucus extracts were found to be approximately 1.5 to 3.0 times lower against fish pathogens and approximately 1.6 to 6.6 folds lower for human pathogens. This preliminary information suggests that the mucus from these fish species may be a source of novel antimicrobial agents for fish and human health related applications.

Enzymatic hydrolysis of esterified diarrhetic shellfish poisoning toxins and pectenotoxins.

Okadaic acid (OA) and dinophysistoxins-1 and -2 (DTX1, DTX2), the toxins responsible for incidents of diarrhetic shellfish poisoning (DSP), can occur as complex mixtures of ester derivatives in both plankton and shellfish. Alkaline hydrolysis is usually employed to release parent OA/DTX toxins, and analyses are conducted before and after hydrolysis to determine the concentrations of nonesterified and esterified toxins. Recent research has shown that other toxins, including pectenotoxins and spirolides, can also exist as esters in shellfish, but these toxins cannot survive alkaline hydrolysis. A promising alternative approach is enzymatic hydrolysis. In this study, two enzymatic methods were developed for the hydrolysis of 7-O-acyl esters, "DTX3," and the carboxylate esters of OA, "diol-esters." Porcine pancreatic lipase induced complete conversion of DTX3 to OA and DTXs within one hour for reference solutions. The presence of mussel tissue matrix reduced the rate of hydrolysis, but an optimized lipase concentration resulted in greater than 95% conversion within four hours. OA-diol-ester was hydrolyzed by porcine liver esterase and was completely converted to OA in less than 30 min, even in the presence of mussel tissue matrix. Esters and OA/DTX toxins were all monitored by LC-MS. Further experiments with pectenotoxin esters indicated that enzymatic hydrolysis could also be applied to esters of other toxins. Enzymatic hydrolysis has excellent potential as an alternative to the conventional alkaline hydrolysis procedure used in the preparation of shellfish samples for the analysis of toxins.