Differential effects of internal tagging depending on depth treatment in Atlantic salmon: a cautionary tale for aquatic animal tag use.

Electronic tags are widespread tools for studying aquatic animal behavior; however, tags risk behavioral manipulation and negative welfare outcomes. During an experiment to test behavioral differences of Atlantic salmon Salmo salar in different aquaculture cage types, including ones expected to elicit deeper swimming behavior, we found negative tagging effects depending on whether cages were depth-modified. In the experiment, data storage tags implanted in Atlantic salmon tracked their depth behavior and survival in unmodified sea-cages and depth-modified sea-cages that forced fish below or into a narrow seawater- or freshwater-filled snorkel tube from a 4 m net roof to the surface. All tagged individuals survived in unmodified cages; however, survival was reduced to 62% in depth-modified cages. Survivors in depth-modified cages spent considerably less time above 4 m than those in unmodified cages, and dying individuals in depth-modified cages tended to position in progressively shallower water. The maximum depth that fish in our study could attain neutral buoyancy was estimated at 22 m in seawater. We calculated that the added tag weight in water reduced this to 8 m, and subtracting the tag volume from the peritoneal cavity where the swim bladder reinflates reduced this further to 4 m. We conclude that the internal tag weight and volume affected buoyancy regulation as well as the survival and behavior of tagged fish. Future tagging studies on aquatic animals should carefully consider the buoyancy-related consequences of internal tags with excess weight in water, and the inclusion of data from dying tagged animals when estimating normal depth behaviors.

Snorkel sea-cage technology decreases salmon louse infestation by 75% in a full-cycle commercial test.

Methods to prevent parasite infestations in farmed fish are becoming widespread, yet tests of their effectiveness often lack commercial relevance and statistical power, which may lead to technology misuse. Here, we examined salmon louse infestation on Atlantic salmon in triplicate commercial snorkel louse barrier and standard cages over a 12 month production cycle. Barrier cages reduced newly settling lice on Atlantic salmon by 75%, with variability in parasite reduction over time depending upon environmental variables. The commercial, triplicate, long-term study design serves as a template to validate performance and detect weaknesses in anti-parasite techniques in fish mariculture.

The effectiveness of hyposaline treatments against host-attached salmon lice.

Understanding how salinity affects marine parasites is vital to understanding their ecology and treatment, particularly for host-parasite systems that traverse marine and freshwater realms such as the globally important Atlantic salmon (Salmo salar), salmon louse (Lepeophtheirus salmonis) system. Growing concerns for wild fish populations, and decreased efficiencies and burgeoning costs of lice treatments for farmed fish has necessitated more environmentally and socially acceptable delousing procedures, such as hyposaline treatments. The effect of brackish water on L. salmonis following primary attachment is largely unknown, with experimental evidence derived mostly from unattached or newly attached copepodids, or adult stages. We aimed to understand how attached lice respond to hyposaline environments to assess effectiveness as a parasite management strategy and to help better define delousing areas used by wild fish. Louse development at 4, 12, 19 and 26 ppt, and survival at 4 ppt, decreased as exposure times increased, but survival was otherwise unaffected. Subjecting salmon to fluctuating, repeat exposures did not influence efficacy. We confirm that free-swimming stages are susceptible, and show that attached copepodids were more tolerant than previously predicted based on experiments on alternate development stages. These results improve our understanding of the utility of hyposaline treatments in aquaculture and self-treating in wild fish, and could apply to other fish-lice parasite systems. Further, these data are important for models predicting host-parasite interactions and can contribute to predictive models on the transmission dynamics of sea lice from farm to wild fish.

Sea lice infestation levels decrease with deeper 'snorkel' barriers in Atlantic salmon sea-cages.

BACKGROUND: Salmon lice (Lepeophtheirus salmonis) are the most important parasites of farmed salmon. Infective larvae position themselves in the upper part of the water column to increase encounter probabilities with potential hosts. Previous studies have shown that a 'snorkel' sea-cage technology protects salmon from infection in surface waters. We tested whether deep snorkels would more effectively reduce lice infestation than shallow snorkels and still uphold adequate conditions for the fish. Five sea-cages (12 m × 12 m) each holding approximately 3000 Atlantic salmon (Salmo salar) (53 ± 10 g) were fitted with snorkels that gave protection from infection for 0, 4, 8, 12 or 16 m. We tested whether reductions in the settlement of new salmon lice copepodids were consistent among four separate infection periods. RESULTS: Lice infestation decreased exponentially with depth in all time periods. Infection levels in shallow snorkels (0 and 4 m) were consistently 4-10 times higher than those in deep snorkels (12 and 16 m). Key welfare and production performance indices were similar across all snorkel depths. CONCLUSION: Deeper snorkels dramatically and consistently reduced infection levels of salmon lice compared with shallow snorkels, without consequences for fish welfare and production performance. Therefore, reducing salmon sea lice encounters using a depth-based barrier is a powerful management tool for salmon farming. © 2017 Society of Chemical Industry.

Exploring the divalent effect in fucosidase inhibition with stereoisomeric pyrrolidine dimers.

Multi-valent inhibitors offer promise for the enhancement of therapeutic compounds across a range of chemical and biological processes. Here, a significant increase in enzyme-inhibition potencies was observed with a dimeric iminosugar-templated fucosidase inhibitor (IC50 = 0.108 µM) when compared to its monovalent equivalent (IC50 = 2.0 µM). Such a gain in binding is often attributed to a "multivalent effect" rising from alternative recapture of the scaffolded binding epitopes. The use of control molecules such as the meso analogue (IC50 = 0.365 µM) or the enantiomer (IC50 = 569 µM), as well as structural analysis of the fucosidase-inhibitor complex, allowed a detailed analysis of the possible mechanism of action, at the molecular level. Here, the enhanced binding affinity of the dimer over the monomer can be attributed to additional interactions in non-catalytic sites as also revealed in the 3-D structure of a bacterial fucosidase inhibitor complex.

A second-generation ferrocene-iminosugar hybrid with improved fucosidase binding properties.

The synthesis and the biological evaluation of a new ferrocenyl-iminosugar conjugate designed for fucosidase inhibitory and anticancer activity is described. The compound showed strong affinity for fucosidase from bovine kidney (Ki=23 nM) and from Bacteroides thetaiotaomicron (Ki=150 nM), displaying a 10-fold tighter binding affinity for these enzymes than the previous analogs. The interaction pattern that improves binding has been evaluated through structural analysis of the inhibitor-enzyme complex. The ferrocenyl-iminosugar exhibits significant anticancer activity on MDA-MB-231 and SK-MEL28 cell lines at 100 µM.

Rational steering of insulin binding specificity by intra-chain chemical crosslinking.

Insulin is a key hormone of human metabolism with major therapeutic importance for both types of diabetes. New insulin analogues with more physiological profiles and better glycemic control are needed, especially analogues that preferentially bind to the metabolic B-isoform of insulin receptor (IR-B). Here, we aimed to stabilize and modulate the receptor-compatible conformation of insulin by covalent intra-chain crosslinking within its B22-B30 segment, using the Cu(I)-catalyzed Huisgen 1,3-dipolar cycloaddition reaction of azides and alkynes. This approach resulted in 14 new, systematically crosslinked insulin analogues whose structures and functions were extensively characterized and correlated. One of the analogues, containing a B26-B29 triazole bridge, was highly active in binding to both IR isoforms, with a significant preference for IR-B. Our results demonstrate the potential of chemistry-driven modulation of insulin function, also shedding new light on the functional importance of hormone's B-chain C-terminus for its IR-B specificity.

In vitro and in vivo comparative and competitive activity-based protein profiling of GH29 α-l-fucosidases.

GH29 α-l-fucosidases catalyze the hydrolysis of α-l-fucosidic linkages. Deficiency in human lysosomal α-l-fucosidase (FUCA1) leads to the recessively inherited disorder, fucosidosis. Herein we describe the development of fucopyranose-configured cyclophellitol aziridines as activity-based probes (ABPs) for selective in vitro and in vivo labeling of GH29 α-l-fucosidases from bacteria, mice and man. Crystallographic analysis on bacterial α-l-fucosidase confirms that the ABPs act by covalent modification of the active site nucleophile. Competitive activity-based protein profiling identified l-fuconojirimycin as the single GH29 α-l-fucosidase inhibitor from eight configurational isomers.

Exploiting the hydrophobic terrain in fucosidases with aryl-substituted pyrrolidine iminosugars.

Fucosidase inhibition shows potential in numerous therapeutic contexts. Substitution of fucose-like iminosugars with hydrophobic "aglycons" yields significant improvements in potency of fucosidase inhibition. Here we have prepared three new 2-aryl-3,4-dihydroxy-5-methylpyrrolidines featuring phenyl substituents in variable orientations with respect to the iminocyclitol core and at various distances from it to explore the key binding interactions that stabilise the enzyme-inhibitor complex. The presence of a triazole linker in one structure resulted in nanomolar inhibition of the fucosidase from bovine kidney (Ki =4.8 nM), thus giving rise to one of the most potent pyrrolidine-type inhibitors of this enzyme known to date.

Historical land-use influences the long-term stream turbidity response to a wildfire.

Wildfires commonly result in an increase in stream turbidity. However, the influence of pre-fire land-use practices on post-fire stream turbidity is not well understood. The Lower Cotter Catchment (LCC) in south-eastern Australia is part of the main water supply catchment for Canberra with land in the catchment historically managed for a mix of conservation (native eucalypt forest) and pine (Pinus radiata) plantation. In January 2003, wildfires burned almost all of the native and pine forests in the LCC. A study was established in 2005 to determine stream post-fire turbidity recovery within the native and pine forest areas of the catchment. Turbidity data loggers were deployed in two creeks within burned native forest and burned pine forest areas to determine turbidity response to fire in these areas. As a part of the study, we also determined changes in bare soil in the native and pine forest areas since the fire. The results suggest that the time, it takes turbidity levels to decrease following wildfire, is dependent upon the preceding land-use. In the LCC, turbidity levels decreased more rapidly in areas previously with native vegetation compared to areas which were previously used for pine forestry. This is likely because of a higher percentage of bare soil areas for a longer period of time in the ex-pine forest estate and instream stores of fine sediment from catchment erosion during post-fire storm events. The results of our study show that the previous land-use may exert considerable control over on-going turbidity levels following a wildfire.

Three dimensional structure of a bacterial α-l-fucosidase with a 5-membered iminocyclitol inhibitor.

Fucosidases, enzymes that cleave fucose from the non-reducing end of a glycan, represent promising medicinal targets reflecting their roles in cancer metastasis, inflammation, host-parasite interactions and the lysosomal storage disorder fucosidosis. The X-ray crystal structures of Bacteroides thetaiotaomicron GH29 α-l-fucosidase (BtFuc2970) in a new crystal form (at a resolution of 1.59Å) and liganded with a 5-membered iminocyclitol inhibitor (1.73Å) are reported herein. The 5-membered iminocyclitol binds in a (3)E conformation, mimicking the proposed (3)H4 half chair transition-state of the enzyme catalysed reaction, and its Ki for BtFuc2970 was determined as 2µM. Structural analysis of fucosidase inhibition through 5-membered iminocyclitols will aid in the rational design of more potent fucosidase inhibitors for treatment of a range of medical conditions.

α-L-fucosidase inhibition by pyrrolidine-ferrocene hybrids: rationalization of ligand-binding properties by structural studies.

Enhanced metabolism of fucose through fucosidase overexpression is a signature of some cancer types, thus suggesting that fucosidase-targetted ligands could play the role of drug-delivery vectors. Herein, we describe the synthesis of a new series of pyrrolidine-ferrocene conjugates, consisting of a L-fuco-configured dihydroxypyrrolidine as the fucosidase ligand armed with a cytotoxic ferrocenylamine moeity. Three-dimensional structures of several of these fucosidase inhibitors reveal transition-state-mimicking (3)E conformations. Elaboration with the ferrocenyl moiety results in sub-micromolar inhibitors of both bovine and bacterial fucosidases, with the 3D structure of the latter revealing electron density indicative of highly mobile alkylferrocene compounds. The best compounds show a strong antiproliferative effect, with up to 100% inhibition of the proliferation of MDA-MB-231 cancer cells at 50 µM.