Divergent responses of pelagic and benthic fish body-size structure to remoteness and protection from humans.

Animal body-size variation influences multiple processes in marine ecosystems, but habitat heterogeneity has prevented a comprehensive assessment of size across pelagic (midwater) and benthic (seabed) systems along anthropic gradients. In this work, we derive fish size indicators from 17,411 stereo baited-video deployments to test for differences between pelagic and benthic responses to remoteness from human pressures and effectiveness of marine protected areas (MPAs). From records of 823,849 individual fish, we report divergent responses between systems, with pelagic size structure more profoundly eroded near human markets than benthic size structure, signifying greater vulnerability of pelagic systems to human pressure. Effective protection of benthic size structure can be achieved through MPAs placed near markets, thereby contributing to benthic habitat restoration and the recovery of associated fishes. By contrast, recovery of the world's largest and most endangered fishes in pelagic systems requires the creation of highly protected areas in remote locations, including on the High Seas, where protection efforts lag.

Shallow subtidal marine benthic communities of Nachvak Fjord, Nunatsiavut, Labrador: A glimpse into species composition and drivers of their distribution.

Marine fjords along the northern Labrador coast of Arctic Canada are influenced by freshwater, nutrients, and sediment inputs from ice fields and rivers. These ecosystems, further shaped by both Atlantic and Arctic water masses, are important habitats for fishes, marine mammals, seabirds, and marine invertebrates and are vital to the Labrador Inuit who have long depended on these areas for sustenance. Despite their ecological and socio-cultural importance, these marine ecosystems remain largely understudied. Here we conducted the first quantitative underwater scuba surveys, down to 12 m, of the nearshore marine ecology of Nachvak Fjord, which is surrounded by Torngat Mountains National Park located in Nunatsiavut, the Indigenous lands claim region of northeastern Canada. Our goal was to provide the Nunatsiavut Government with a baseline of the composition and environmental influences on the subtidal community in this isolated region as they work towards the creation of an Indigenous-led National Marine Conservation Area that includes Nachvak Fjord. We identified four major benthic habitat types: (1) boulders (2) rocks with sediment, (3) sediment with rocks, and (4) unconsolidated sediments, including sand, gravel, and cobble. Biogenic cover (e.g., kelp, coralline algae, and sediment) explained much of the variability in megabenthic invertebrate community structure. The kelp species Alaria esculenta, Saccharina latissima, and Laminaria solidungula dominated the boulder habitat outside of the fjord covering 35%, 13%, and 11% of the sea floor, respectively. In contrast, the middle and inner portions of the fjord were devoid of kelp and dominated by encrusting coralline algae. More diverse megabenthic invertebrate assemblages were detected within the fjord compared to the periphery. Fish assemblages were depauperate overall with the shorthorn sculpin, Myoxocephalus scorpius, and the Greenland cod, Gadus ogac, dominating total fish biomass contributing 64% and 30%, respectively. Understanding the composition and environmental influences within this fjord ecosystem not only contributes towards the protection of this ecological and culturally important region but serves as a baseline in a rapidly changing climatic region.

Sharks are the preferred scraping surface for large pelagic fishes: Possible implications for parasite removal and fitness in a changing ocean.

Mutualistic and commensal interactions can have significant positive impacts on animal fitness and survival. However, behavioural interactions between pelagic animals living in offshore oceanic environments are little studied. Parasites can negatively effect the fitness of their hosts by draining resources and diverting energy from growth, reproduction, and other bodily functions. Pelagic fishes are hosts to a diverse array of parasites, however their environment provides few options for removal. Here we provide records of scraping behaviour of several pelagic teleost species, a behaviour that is likely used for parasite removal. These records span three ocean basins and, to the best of our knowledge, include the first records of scraping interactions involving tunas, blue sharks, and mako sharks as well as the first records of intraspecific scraping. We found that scrapers preferred scraping their head, eyes, gill cover, and lateral surfaces, areas where parasites are commonly found and where damage would likely have a substantial impact on fitness. Scraper species varied in their scraping preferences with tunas scraping mostly on the posterior caudal margins of sharks and occasionally conspecifics, while rainbow runner scraped in more varied locations on both sharks and conspecifics. Lengths of scrapers and scrapees were positively correlated and fish scraping on sharks were larger than those scraping on conspecifics, suggesting that risk of predation may be a limiting factor. We show that pelagic teleosts prefer to scrape on sharks rather than conspecifics or other teleosts and suggest that this behaviour may have a positive impact on teleost fitness by reducing parasite loads. The decline of shark populations in the global ocean and the reduction in mean size of many species may limit these interactions, eroding possible fitness benefits associated with this behaviour, and consequently placing more pressure on already highly targeted and vulnerable species.

Release of Staphylococcus aureus extracellular vesicles and their application as a vaccine platform.

Secretion of extracellular vesicles (EVs), a process common to eukaryotes, archae, and bacteria, represents a secretory pathway that allows cell-free intercellular communication. Microbial EVs package diverse proteins and influence the host-pathogen interaction, but the mechanisms underlying EV production in Gram-positive bacteria are poorly understood. Here we show that EVs purified from community-associated methicillin-resistant Staphylococcus aureus package cytosolic, surface, and secreted proteins, including cytolysins. Staphylococcal alpha-type phenol-soluble modulins promote EV biogenesis by disrupting the cytoplasmic membrane; whereas, peptidoglycan cross-linking and autolysin activity modulate EV production by altering the permeability of the cell wall. We demonstrate that EVs purified from a S. aureus mutant that is genetically engineered to express detoxified cytolysins are immunogenic in mice, elicit cytolysin-neutralizing antibodies, and protect the animals in a lethal sepsis model. Our study reveals mechanisms underlying S. aureus EV production and highlights the usefulness of EVs as a S. aureus vaccine platform.

First quantification of subtidal community structure at Tristan da Cunha Islands in the remote South Atlantic: from kelp forests to the deep sea.

Tristan da Cunha Islands, an archipelago of four rocky volcanic islands situated in the South Atlantic Ocean and part of the United Kingdom Overseas Territories (UKOTs), present a rare example of a relatively unimpacted temperate marine ecosystem. We conducted the first quantitative surveys of nearshore kelp forests, offshore pelagic waters and deep sea habitats. Kelp forests had very low biodiversity and species richness, but high biomass and abundance of those species present. Spatial variation in assemblage structure for both nearshore fish and invertebrates/algae was greatest between the three northern islands and the southern island of Gough, where sea temperatures were on average 3-4o colder. Despite a lobster fishery that provides the bulk of the income to the Tristan islands, lobster abundance and biomass are comparable to or greater than many Marine Protected Areas in other parts of the world. Pelagic camera surveys documented a rich biodiversity offshore, including large numbers of juvenile blue sharks, Prionace glauca. Species richness and abundance in the deep sea is positively related to hard rocky substrate and biogenic habitats such as sea pens, crinoids, whip corals, and gorgonians were present at 40% of the deep camera deployments. We observed distinct differences in the deep fish community above and below ~750 m depth. Concurrent oceanographic sampling showed a discontinuity in temperature and salinity at this depth. While currently healthy, Tristan's marine ecosystem is not without potential threats: shipping traffic leading to wrecks and species introductions, pressure to increase fishing effort beyond sustainable levels and the impacts of climate change all could potentially increase in the coming years. The United Kingdom has committed to protection of marine environments across the UKOTs, including Tristan da Cunha and these results can be used to inform future management decisions as well as provide a baseline against which future monitoring can be based.

Antibodies to Staphylococcus aureus capsular polysaccharides 5 and 8 perform similarly in vitro but are functionally distinct in vivo.

The capsular polysaccharide (CP) produced by Staphylococcus aureus is a virulence factor that allows the organism to evade uptake and killing by host neutrophils. Polyclonal antibodies to the serotype 5 (CP5) and type 8 (CP8) capsular polysaccharides are opsonic and protect mice against experimental bacteremia provoked by encapsulated staphylococci. Thus, passive immunotherapy using CP antibodies has been considered for the prevention or treatment of invasive antibiotic-resistant S. aureus infections. In this report, we generated monoclonal antibodies (mAbs) against S. aureus CP5 or CP8. Backbone specific mAbs reacted with native and O-deacetylated CPs, whereas O-acetyl specific mAbs reacted only with native CPs. Reference strains of S. aureus and a selection of clinical isolates reacted by colony immunoblot with the CP5 and CP8 mAbs in a serotype-specific manner. The mAbs mediated in vitro CP type-specific opsonophagocytic killing of S. aureus strains, and mice passively immunized with CP5 mAbs were protected against S. aureus bacteremia. Neither CP8-specific mAbs or polyclonal antibodies protected mice against bacteremia provoked by serotype 8 S. aureus clinical isolates, although these same antibodies did protect against a serotype 5 S. aureus strain genetically engineered to produce CP8. We detected soluble CP8 in culture supernatants of serotype 8 clinical isolates and in the plasma of infected animals. Serotype 5 S. aureus released significantly less soluble CP5 in vitro and in vivo. The release of soluble CP8 by S. aureus may contribute to the inability of CP8 vaccines or antibodies to protect against serotype 8 staphylococcal infections.

Protective efficacy of a novel alpha hemolysin subunit vaccine (AT62) against Staphylococcus aureus skin and soft tissue infections.

Alpha hemolysin (Hla) is a pore-forming toxin produced by most Staphylococcus aureus isolates. Hla is reported to play a key role in the pathogenesis of staphylococcal infections, such as skin and soft tissue infection, pneumonia, and lethal peritonitis. This study makes use of a novel recombinant subunit vaccine candidate (AT62) that was rationally designed based on the Hla heptameric crystal structure. AT62 comprises a critical structural domain at the N terminus of Hla, and it has no inherent toxic properties. We evaluated the efficacy of AT62 in protection against surgical wound infection and skin and soft tissue infection. Mice were vaccinated on days 0, 14, and 28 with 20µg AT62 or bovine serum albumin (BSA) mixed with Sigma adjuvant system®. Mice immunized with AT62 produced a robust antibody response against native Hla. In the surgical wound infection model, mice immunized with AT62 and challenged with a USA300 S. aureus strain showed a significantly reduced bacterial burden in the infected tissue compared to animals given BSA. Similarly, mice passively immunized with rabbit IgG to AT62 showed reduced wound infection and tissue damage. Subcutaneous abscess formation was not prevented by immunization with AT62. However, in a skin necrosis infection model, immunization with the AT62 vaccine resulted in smaller lesions and reduced mouse weight loss compared to controls. Although AT62 immunization reduced tissue necrosis, it did not reduce the bacterial burdens in the lesions compared to controls. Our data indicate that AT62 may be a valuable component of a multivalent vaccine against S. aureus.

Simplifying metal-'ate' chemistry: formation and comprehensive characterisation of a homo-metallic amido lithiate complex.

Addition of TMEDA to lithium allyl-1-naphthylamide in a 1 : 1 ratio afforded the novel amido lithium/lithiate complex, [Li(TMEDA)2][Li{N(1-naph)(CH2CH[double bond, length as m-dash]CH2)}2]. (7)Li NMR and computational calculations indicate this model to be representative of the solution state, with (1)H and (13)C NMR showing a 1,3-sigmatropic rearrangement. [Li{N(1-naph)(CH2CH[double bond, length as m-dash]CH2)}·Et2O]2, and [Li{N(1-naph) (CH2CH[double bond, length as m-dash]CH2)}·PMDETA] , are presented for comparison.

Preclinical Efficacy of Clumping Factor A in Prevention of Staphylococcus aureus Infection.

UNLABELLED: Treatment of Staphylococcus aureus infections has become increasingly difficult because of the emergence of multidrug-resistant isolates. Development of a vaccine to prevent staphylococcal infections remains a priority. To determine whether clumping factor A (ClfA) is a good target protein for inclusion in a multivalent vaccine, we evaluated its efficacy in a variety of relevant staphylococcal infection models, challenging with different S. aureus strains. ClfA adsorbed to Alhydrogel and mixed with Sigma Adjuvant System was more immunogenic and stimulated a more robust Th17 response than ClfA administered with alum alone. ClfA immunization induced the production of functional antibodies in rabbits and mice that blocked S. aureus binding to fibrinogen and were opsonic for S. aureus strains that produced little or no capsular polysaccharide. Mice immunized with ClfA showed a modest reduction in the bacterial burden recovered from subcutaneous abscesses provoked by S. aureus USA300 strain LAC. In addition, the ClfA vaccine reduced lethality in a sepsis model following challenge with strain Newman, but not ST80. Vaccination with ClfA did not protect against surgical wound infection, renal abscess formation, or bacteremia. Passive immunization with antibodies to ClfA did not protect against staphylococcal bacteremia in mice or catheter-induced endocarditis in rats. Some enhancement of bacteremia was observed by ClfA immunization or passive administration of ClfA antibodies when mice were challenged by the intraperitoneal route. Although rodent models of staphylococcal infection have their limitations, our data do not support the inclusion of ClfA in an S. aureus multivalent vaccine. IMPORTANCE: Antibiotics are often ineffective in eradicating Staphylococcus aureus infections, and thus, a preventative vaccine is sorely needed. Two single-component vaccines and two immunoglobulin preparations failed to meet their designated endpoints in phase III clinical trials. Importantly, recipients of an S. aureus surface protein (iron surface determinant B) vaccine who developed a staphylococcal infection experienced a higher rate of multiorgan failure and mortality than placebo controls, raising safety concerns. Multicomponent S. aureus vaccines have now been generated, and several include surface protein clumping factor A (ClfA). We immunized mice with ClfA and generated a robust T cell response and serum antibodies that were functional in vitro. Nonetheless, ClfA was not protective in a number of relevant animal models of S. aureus infection, and high levels of ClfA antibodies enhanced bacteremia when mice were challenged with community-acquired methicillin-resistant S. aureus strains. Evidence supporting ClfA as a vaccine component is lacking.

Synchrotron far infrared spectroscopy of the ground, ν5, and ν15 states of thiirane.

The high-resolution (0.001 cm(-1)) spectrum of thiirane has been recorded at the far-infrared beamline at the Australian synchrotron between 760-400 cm(-1) and 170-10 cm(-1). Ro-vibrational transitions of the highly Coriolis coupled ν(5) (628.1 cm(-1)) and ν(15) (669.7 cm(-1)) fundamentals, as well as pure rotational far-IR transitions have been assigned, and rotational, centrifugal distortion, and Coriolis interaction parameters determined. ν(15) gains the vast majority of its intensity from an interesting Coriolis intensity stealing mechanism, which is also outlined.

High-resolution Fourier-transform infrared spectroscopy of the ν6 and Coriolis perturbation allowed ν10 modes of ketenimine.

High-resolution FTIR spectra of the short lived species ketenimine have been recorded in the region 700-1300 cm(-1) and over 1500 transitions of the ν(10) and ν(6) modes have been assigned. Effective rotational and centrifugal distortion parameters for the v(10) = 1 and v(6) = 1 (excluding K(a) = 5) states were determined by co-fitting transitions, and treating strong a- and c-axis Coriolis interactions between them. Other perturbations attributed to interactions with the v(8) = 2 and v(12) = 1 + v(8) = 1 dark-states were also observed and treated. The ν(10) transitions are predicted to be inherently very weak, but are enhanced by an intensity stealing effect with the highly IR active ν(6) mode. A mechanism for this intensity stealing in ketenimine is also detailed.

High-resolution Fourier-transform infrared spectroscopy of the Coriolis coupled ground state and ν7 mode of ketenimine.

High resolution FTIR spectra of the short lived species ketenimine have been recorded in the regions 390-1300 cm(-1) and 20-110 cm(-1) using synchrotron radiation. Two thousand six hundred sixty transitions of the ν(7) band centered at 693 cm(-1) and 126 far-IR rotational transitions have been assigned. Rotational and centrifugal distortion parameters for the ν(7) mode were determined and local Fermi and b-axis Coriolis interactions with 2ν(12) are treated. A further refinement of the ground state, ν(12) and ν(8) parameters was also achieved, including the treatment of previously unrecognized ac-axis and ab-axis second order perturbations to the ground state.

High-resolution FTIR spectroscopy of the ν8 and Coriolis perturbation allowed ν12 bands of ketenimine.

High resolution FTIR spectra have been recorded in the region 250-770 cm(-1) using synchrotron radiation and over 2000 transitions to the ν(8) and ν(12) states of the short lived species ketenimine have been assigned. Ground state combination differences combined with published microwave transitions were used to refine the constants for the ground vibrational state. Rotational and centrifugal distortion parameters for the v(8) = 1 and v(12) = 1 levels were determined by co-fitting transitions, and treating a strong a-axis Coriolis interaction. Selection rules for the observed ν(12) transitions indicate that they arise solely from "perturbation allowed" intensity resulting from this Coriolis interaction.

Overview of high-resolution infrared measurement and analysis for atmospheric monitoring of halocarbons.

The current state of the art in recording and analyzing rotationally resolved vibration-rotation bands of atmospheric pollutant halocarbon species is reviewed. It is shown that in order to obtain molecular constants of sufficient accuracy to simulate the vibration-rotation structure over the range of atmospheric temperatures, it is necessary to obtain spectra at a range of temperatures using static cooling cells, supersonic jet expansions, and collisional cooling devices; employ sophisticated pattern recognition and analysis software; assign and fit spectral perturbations; and use spectral simulation and digital spectral subtraction (SASSI) to further simplify spectral bands for analysis. To demonstrate the techniques, an analysis of the ν(5) band of CH(37)ClF(2) in natural abundance is presented.

A spectroscopic study of nicotine analogue 2-phenylpyrrolidine (PPD) using resonant two-photon ionization (R2PI), microwave, and 2D NMR techniques.

Conformational preferences of the nicotine analogue 2-phenylpyrrolidine (PPD) have been studied in both gaseous and solution phases. Theoretical calculations at the MP2 and B3LYP levels point to 5-6 stable conformers which differ in three degrees of conformational freedom; torsion between the two rings, inversion at the pyrrolidine (PY) amine, and PY ring puckering, characterized using the Cremer-Pople definition for pseudorotation. Only one conformer has a trans arrangement between the amino hydrogen and the phenyl substituent. It is 6-8 kJ mol(-1) more stable than the cis conformers, has a perpendicular ring arrangement, and puckers at the nitrogen atom--similar to structures reported for nicotine. Resonant two-photon ionization (R2PI) data, including hole burn spectra, indicate only one conformer is present in the free jet expansion, and band contour analysis suggests assignment to the trans conformer. Confirmation was provided by microwave spectroscopy. Fifty-seven lines measured in the 48-72 GHz region were assigned to 206 b-type transitions and fitted to yield rotational constants within 2 MHz of MP2 values predicted for the trans conformer. The solution-phase conformers of PPD were studied using 1D and 2D (1)H NMR spectroscopy and solvent-based theoretical calculations. In marked contrast to the gas phase, NMR data reveals only cis conformers present in solution. Calculations confirm increased stability for these conformers when placed in simulated chloroform or water environments. Solvent molecules are believed to disrupt a crucial N...H(ortho) stabilizing interaction present within the trans conformer.

Resonant 2-photon ionization study of the conformation and the binding of water molecules to 2-phenylethanethiol (PhCH2CH2SH).

The structures of 2-phenylethanethiol (PET, PhCH(2)CH(2)SH) and its 1:1 water clusters have been studied using resonant two-photon ionization spectroscopy including band contour analysis and UV-UV holeburning, combined with extensive ab initio calculations on ground and excited states. The most populated conformer, labeled Ggpi, has a gauche arrangement about the SCCC and HSCC bonds that permits a stabilizing SH...dpi type of hydrogen bond. The other observed conformer, Ag, is anti with respect to the SCCC bond. In the dominant 1:1 water cluster, a water molecule binds to the Ggpi conformer via an OH...S hydrogen bond and two significant CH...O interactions. There is also evidence for water binding to conformer Ag with a similar arrangement, and for a second Ggpi cluster where water inserts between the SH and the aromatic ring. The additional interactions to the water molecules result in net D(e) binding energies approximately double those resulting from a single thiol-water hydrogen bond. The (1)(pi,pi(*)) excited state lifetimes in the bare molecules are very short because of internal conversion to a dissociative (1)(n,pi(*)) state related to the thiol. In the dominant Gw(1) cluster, the lifetime is significantly increased from <1 to approximately 4 ns. Hydrogen bonding to the thiol, which raises the energy of the dissociative (1)(n,pi(*)) state, accounts for this behavior.

Interactions between the chloride anion and aromatic molecules: infrared spectra of the Cl- -C6H5CH3, Cl- -C6H5NH2 and Cl- -C6H5OH complexes.

The Cl- -C6H5CH3*Ar, Cl- -C6H5NH2*Ar, and Cl- -C6H5OH*Ar anion complexes are investigated using infrared photodissociation spectroscopy and ab initio calculations at the MP2/aug-cc-pVDZ level. The results indicate that for Cl- -C6H5NH2 and Cl- -C6H5OH, the Cl- anion is attached to the substituent group by a single near-linear hydrogen bond. For Cl--C6H5CH3, the Cl- is attached to an ortho-hydrogen atom on the aromatic ring and to a hydrogen atom on the methyl group by a weaker hydrogen bond. The principal spectroscopic consequence of the hydrogen-bonding interaction in the three complexes is a red-shift and intensity increase for the CH, NH, and OH stretching modes. Complexities in the infrared spectra in the region of the hydrogen-bonded XH stretch band are associated with Fermi resonances between the hydrogen-stretching vibrational modes and bending overtone and combination levels. There are notable correlations between the vibrational red-shift, the elongation of the H-bonded XH group, and the proton affinity of the aromatic molecule's conjugate base.

Interplanar torsion in the S1<--S0 electronic spectrum of jet cooled 1-phenylimidazole.

The S(1)<--S(0) transition of 1-phenylimidazole (1PI) has been studied in a supersonic jet expansion by resonant two-photon ionization. The origin band at 36 075 cm(-1) is accompanied by a low frequency progression associated with torsion about the bond connecting phenyl and imidazole groups. Torsional potentials have been determined for both states. In S(0), phi(min)=37.2+/-0.5 degrees and the planar barrier is 339+/-20 cm(-1), while in S(1), phi(min)=17.6+/-0.5 degrees and the planar barrier is 57+/-2 cm(-1). The transition moment alignment is observed to be consistent with an excited state of L(b) character, in spite of the "off-axis" conjugation provided by the imidazole ring. These results are compared with ab initio calculations on both states, performed using Hartree-Fock, Møller-Plesset second-order perturbation, density functional theory with the Becke3-Lee-Yang-Parr functional, time-dependent density functional theory, configuration interaction singles, and complete active space self-consistent field methods. Solution-phase UV spectra of neutral and protonated 1PI are also reported.