Evidence for marine origin and microbial-viral habitability of sub-zero hypersaline aqueous inclusions within permafrost near Barrow, Alaska.

Cryopegs are sub-surface hypersaline brines at sub-zero temperatures within permafrost; their global extent and distribution are unknown. The permafrost barrier to surface and groundwater advection maintains these brines as semi-isolated systems over geological time. A cryopeg 7 m below ground near Barrow, Alaska, was sampled for geochemical and microbiological analysis. Sub-surface brines (in situtemperature of -6 °C, salinity of 115 ppt), and an associated sediment-infused ice wedge (melt salinity of 0.04 ppt) were sampled using sterile technique. Major ionic concentrations in the brine corresponded more closely to other (Siberian) cryopegs than to Standard seawater or the ice wedge. Ionic ratios and stable isotope analysis of water conformed to a marine or brackish origin with subsequent Rayleigh fractionation. The brine contained ∼1000× more bacteria than surrounding ice, relatively high viral numbers suggestive of infection and reproduction, and an unusually high ratio of particulate to dissolved extracellular polysaccharide substances. A viral metagenome indicated a high frequency of temperate viruses and limited viral diversity compared to surface environments, with closest similarity to low water activity environments. Interpretations of the results underscore the isolation of these underexplored microbial ecosystems from past and present oceans.

Two new species of Paraorygmatobothrium Ruhnke, 1994 (Tetraphyllidea: Phyllobothriidae) from the smooth-hound Mustelus mustelus (L.) and the gummy shark M. antarcticus Günther (Carcharhiniformes: Triakidae).

Two new tetraphyllidean species, Paraorygmatobothrium bai n. sp. and P. rodmani n. sp., are described from the smooth-hound shark Mustelus mustelus (Linnaeus) and the gummy shark M.s antarcticus Günther, respectively. The two species are generally consistent in morphology with the nine existing species of Paraorygmatobothrium Ruhnke, 1994. P. bai n. sp. and P. rodmani n. sp. differ from the existing species of Paraorygmatobothrium in exhibiting gravid proglottids on the strobila. P. bai n. sp. differs from P. rodmani n. sp. in testicular shape and number, in addition to significant differences in body length, and terminal and subterminal proglottis length to width ratios. Description of these two new species increases the known number of species of Paraorygmatobothrium to 11.

Orectolobicestus n. g. (Cestoda: Tetraphyllidea), with the description of five new species and the transfer of Phyllobothrium chiloscyllii to the new genus.

ORECTOLOBICESTUS N. G. (CESTODA: Tetraphyllidea) is erected for six cestode species parasitising bamboo sharks (Orectolobiformes: Chiloscyllium). Members of this genus differ from all other phyllobothriid genera in possessing modified maisiform spinitriches on their distal bothridial surfaces. In addition, they are easily recognised in that they share the following unique combination of characters: their vitelline fields are interrupted by the ovary, their necks are scutellate, and their bothridia bear an apical sucker and marginal loculi. Five new species of Orectolobicestus are described, including O. tyleri n. sp. from Chiloscyllium punctatum off Borneo, O. lorettae n. sp. from C. cf. punctatum off Australia, O. mukahensis n. sp. and O. kelleyae n. sp. from C. indicum off Borneo, and O. randyi n.sp. from C. hasselti also from off Borneo. In addition, Phyllobothrium chiloscyllii Subhapradha, 1955 is transferred to the new genus. O. chiloscyllii (Subhapradha, 1955) n. comb. is readily distinguished from all five new species in its greater total length. In addition to a number of proglottid features, O. kelleyae n. sp. and O. randyi n. sp. clearly differ from the other three new species in their possession of trifid, rather than fully serrate, spinitriches on their proximal bothridial surfaces. The latter two species conspicuously differ from one another in total number of proglottids (11-21 vs 27-38). O. tyleri n. sp. generally has fewer proglottids than O. lorrettae n. sp. (7-17 vs 13-23) and, like O. mukahensis n. sp., possesses scutes that are spathate rather than elongate. O. tyleri n. sp. is readily distinguished from O. mukahensis n. sp. in its possession of fewer proglottids (7-17 vs. 19-29). The five new species of Orectolobicestus share derived bothridial microthrix features with Phyllobothrium squali Yamaguti, 1952, Thysanocephalum sp., Orygmatobothrium sp., Ruhnkecestus Caira & Durkin, 2006 and species of Paraorygmatobothrium Ruhnke, 1994. Among these taxa, Orectolobicestus most closely resembles Paraorygmatobothrium and Ruhnkecestus in its vitelline fields being interrupted by the ovary and the possession of a scutellate neck.

Effect of pulse shape on the efficiency of multiphoton processes: implications for biological microscopy.

The effects of spectral shape on two photon fluorescence excitation are investigated experimentally using an acousto-optic pulse shaper to modify femtosecond pulses from a Ti:sapphire laser. By using different spectral window shapes, we find that the measured two photon efficiency can vary by a factor of 2 for differently shaped spectra with the same full width at half maximum. We find that these effects are described well by a simple model assuming transform-limited pulses. The fact that even small changes in the spectral wings can significantly affect the efficiency of nonlinear processes has implications for biological multiphoton imaging, where it may be desirable to minimize sample exposure to radiation and maximize fluorescence or harmonic efficiency. © 1999 Society of Photo-Optical Instrumentation Engineers.

Analysis of chimeric spinach/cyanobacterial CP43 mutants of Synechocystis sp. PCC 6803: the chlorophyll-protein CP43 affects the water-splitting system of Photosystem II.

Mutants of the cyanobacterium Synechocystis sp. PCC 6803 have been generated in which parts of psbC (the gene encoding the Photosystem II chlorophyll-protein CP43) have been replaced with the homologous gene fragment from spinach. Upon the replacement of all but the 3' 84 bp of the cyanobacterial psbC gene with the homologous fragment from spinach, an obligate photoheterotrophic mutant was generated. Two photoautotrophic derivatives of this mutant were made reincorporating 3' cyanobacterial sequences back into the spinach psbC gene of the mutant. These two mutants are similar to each other, carrying a chimeric CP43 with the N-terminal half from spinach. These mutants are photosynthetically active at a rate of about half that of wild type, which correlates with a decreased Photosystem II/chlorophyll ratio in these mutants. Thylakoids from the chimeric mutants contain a CP43 protein which migrates slightly more slowly on SDS-polyacrylmide gels than the native Synechocystis CP43. Interestingly, these mutants show significant shifts in thermoluminescence peaks, reflecting altered thermodynamic properties of the back reaction between the acceptor side and the water-splitting system. On the basis of the oscillations of these shifts with number of flashes, we conclude that S2 is stabilized and S3 is destabilized in these mutants. This represents evidence for an involvement of CP43 in events associated with water splitting.

The psbC start codon in Synechocystis sp. PCC 6803.

The translation start codon for psbC, the gene encoding CP43, a chlorophyll-binding protein of photosystem II, has been identified for the cyanobacterium Synechosystis sp. PCC 6803 using site-directed mutagenesis. An AUG codon, about 50 bases upstream from the end of psbD-I had previously been assumed to be the translation start site of psbC. However, the fact that the AUG codon is not present in psbC from several other organisms, whereas a GUG codon 14 bases upstream from the end of psbD-I is strictly conserved suggests that CP43 translation starts at the latter codon. Mutation of GUG, but not of AUG, led to a loss of CP43 and photoautotrophic growth, indicating that the GUG codon is the sole initiation site for translation of the CP43 protein in Synechocystis sp. PCC 6803.