Differential Diagnosis Of Mixed Haplosporidium Costale And Haplosporidium Nelsoni Infections In The Eastern Oyster, Crassostrea Virginica, Using Dna Probes

Haplosporidium costale and Haplosporidium nelsoni are morphologically similar pathogens of the eastern oyster Crassostrea virginica. In the absence of the spore stage, infections of the two species are extremely difficult, if not impossible, to distinguish using traditional light microscopy of stained tissue sections. Species-specific molecular diagnostics were developed for H. costale from the small subunit ribosomal DNA (SSU rDNA) sequence. The polymerase chain reaction (PCR) primers amplified a 557 base pair (bp) region of the H. costale SSU rDNA, but did not amplify DNA from oyster (C. virginica) or from six other haplosporidans (H. nelsoni, H. louisiana, H. lusitanicum, Minchinia teredinis, M. chitonis, or M. tapetis). The DNA probe was used with in sim hybridizations of oyster tissue sections to visualize H. costale plasmodia and prespore stages; it did not hybridize with oyster (C. virginica) or other haplosporidans (H. nelsoni, H. louisiana, or Minchinia teredinis). DNA-based diagnostics for H. costale, in conjunction with molecular tools previously developed for H. nelsoni, have overcome limitations of histological examination. From in situ hybridizations using both probes, some Virginia oysters previously diagnosed with H. costale were found to have mixed infections consisting of approximately 80 to 90% H. costale plasmodia and 10 to 20% H. nelsoni plasmodia, Plasmodia of H. costale were not found in epithelial tissue, only in connective tissue. In addition, use of the DNA probe confirmed the presence of H. costale plasmodia in Virginia oysters collected in the fall, an unprecedented seasonality for an advanced H. costale infection

Detection Of Haplosporidium-nelsoni (Haplosporidia, Haplosporidiidae) In Oysters By PCR Amplification

Haplosporidium nelsoni is a protistan pathogen of the eastern oyster Crassostrea virginica, and has contributed to the decline of the oyster population in the Chesapeake Bay. From comparison of the sequence data of the 16S-like rDNA of H. nelsoni with those of Minchinia teredinis and other related organisms, 2 oligonucleotides which were specific to H. nelsoni and suitable for use as PCR primers were identified. These primers amplified a 564 base pair fragment of the small subunit (SSU) rRNA gene of H. nelsoni, but did not amplify genomic oyster DNA or the SSU rRNA genes of the haplosporidians Haplosporidium costale, Haplosporidium louisiana, or M. teredinis. The PCR primers were able to detect the H. nelsoni SSU rDNA from 50 ng of infected oyster genomic DNA or from 10 fg of cloned H. nelsoni SSU rDNA. The ability of the PCR primers to diagnose H. nelsoni-infected oysters was compared to the established techniques of hemolymph settlement analysis in Farley chambers and histological examination from a sample of 20 oysters. Hemolymph settlement analysis detected infection in 10 oysters and histology revealed infections in 11 oysters. PCR amplification of DNA from hemolymph initially detected infections in 15 oysters and reamplification of the PCR products detected an additional 4 infections. PCR amplification is a more sensitive diagnostic assay for H. nelsoni than traditional techniques

Haplosporidium Costale (Seaside Organism), A Parasite Of The Eastern Oyster, Is Present In Long Island Sound

A haplosporidian parasite, Haplosporidium costale (seaside organism or SSO), is associated with high mortalities of eastern oysters (Crassostrea virginica) in seaside bays of Virginia and Maryland. Its presence in Long Island Sound has been tentatively suggested in several publications for the last 50 y. Positive identification of H. costale and differentiation from another haplosporidian parasite, Haplosporidium nelsoni (MSX), from histological sections is difficult and requires the presence of spores. We detected H. costale spores in 4 out of 5010 (0.08%) oysters collected from Long Island Sound in 1997-1999. In situ hybridization using an oligonucleotide DNA probe designed to detect small subunit ribosomal DNA from Virginia\u27s H. costale reacted positively with tentative H. costale plasmodia in 5 oysters from Long Island Sound. In each case there was a coinfection of H. nelsoni. In Virginia and Maryland, H. costate has historically sporulated in all infected animals in May-June. In Long Island Sound, the rare sporulating cases were detected in October-December, suggesting a different infection cycle

Investigating The Life Cycle Of Haplosporidium nelsoni (MSX)

Attempts to decipher the life cycle of Haplosporidium nelsoni began almost immediately after it was identified as the pathogen causing MSX disease in eastern oysters, Crassostrea virginica. But transmission experiments failed and the spore stage, characteristic of haplosporidans, was extremely rare. Researchers concluded that another host was involved: an intermediate host in which part of the life cycle was produced, or-if the oyster was an accidental host-an alternate host that produces infective elements. A later finding that spores were found more often in spat (\u3c 1 y old) than in adults revived the idea of direct transmission between oysters. The new findings and the availability of molecular diagnostics led us to revive life cycle investigations. Over several years, oyster spat were examined for spores and searched for H. nelsoni in potential non-oyster hosts using both histological and polymerase chain reaction (PCR) methodologies. Although spores occurred in a high proportion of spat with advanced infections, it was concluded that they were unlikely to be a principal source of infective elements because naive oysters used as sentinels to assess infection pressure became highly infected even after native oysters developed resistance, and infected spat could no longer be found. A histological survey of zooplankton and small bivalves in Delaware Bay found few recognizable parasites and nothing resembling a haplosporidan. A subsequent PCR study of water, sediment, and macro-invertebrates from Chesapeake, Delaware, and Oyster bays resulted in many positive samples, but in situ hybridization failed to identify any recognizable structures. PCR analysis of potential intermediate hosts for other molluscan pathogens has also resulted in many species yielding positive results but required in situ hybridization to verify infections. It is suggested that any future search for a nonoyster host of H. nelsoni be conducted in a relatively confined system and/or target specific phyla, strategies that have been successful in other life cycle studies. It is noted that candidate phyla could include those known to host haplosporidans and species whose abundance or distribution may have changed in concert with outbreaks of MSX disease in the northeastern United States in recent years

Seasonal patterns of fine root production and turnover in a mature rubber tree (Hevea brasiliensis Müll. Arg.) stand- differentiation with soil depth and implications for soil carbon stocks

Fine root dynamics is a main driver of soil carbon stocks, particularly in tropical forests, yet major uncertainties still surround estimates of fine root production and turnover. This lack of knowledge is largely due to the fact that studying root dynamics in situ, particularly deep in the soil, remains highly challenging. We explored the interactions between fine root dynamics, soil depth, and rainfall in mature rubber trees (Hevea brasiliensis Müll. Arg.) exposed to sub-optimal edaphic and climatic conditions. A root observation access well was installed in northern Thailand to monitor root dynamics along a 4.5 m deep soil profile. Image-based measurements of root elongation and lifespan of individual roots were carried out at monthly intervals over 3 years. Soil depth was found to have a significant effect on root turnover. Surprisingly, root turnover increased with soil depth and root half-life was 16, 6–8, and only 4 months at 0.5, 1.0, 2.5, and 3.0 m deep, respectively (with the exception of roots at 4.5 m which had a half-life similar to that found between depths of 1.0 and 2.5 m). Within the first two meters of the soil profile, the highest rates of root emergence occurred about 3 months after the onset of the rainy season, while deeper in the soil, root emergence was not linked to the rainfall pattern. Root emergence was limited during leaf flushing (between March and May), particularly within the first two meters of the profile. Between soil depths of 0.5 and 2.0 m, root mortality appeared independent of variations in root emergence, but below 2.0 m, peaks in root emergence and death were synchronized. Shallow parts of the root system were more responsive to rainfall than their deeper counterparts. Increased root emergence in deep soil toward the onset of the dry season could correspond to a drought acclimation mechanism, with the relative importance of deep water capture increasing once rainfall ceased. The considerable soil depth regularly explored by fine roots, even though significantly less than in surface layers in terms of root length density and biomass, will impact strongly the evaluation of soil carbon stocks

Introducing dip pen nanolithography as a tool for controlling stem cell behaviour: unlocking the potential of the next generation of smart materials in regenerative medicine (vol 10, pg 1662, 2010)

Correction for ‘Introducing dip pen nanolithography as a tool for controlling stem cell behaviour: unlocking the potential of the next generation of smart materials in regenerative medicine’ by Judith M. Curran et al., Lab Chip, 2010, 10, 1662–1670.</p

Search for the Θ+\Theta^+ pentaquark in the reaction γd→pK−K+n\gamma d \to p K^- K^+ n

A search for the \thp in the reaction $\gamma d \to pK^-K^+n$ was completed using the CLAS detector at Jefferson Lab. A study of the same reaction, published earlier, reported the observation of a narrow \thp resonance. The present experiment, with more than 30 times the integrated luminosity of our earlier measurement, does not show any evidence for a narrow pentaquark resonance. The angle-integrated upper limit on \thp production in the mass range of 1.52 to 1.56 GeV/c$^2$ for the $\gamma d \to pK^-\Theta^+$ reaction is 0.3 nb (95% CL). This upper limit depends on assumptions made for the mass and angular distribution of \thp production. Using \lamstar production as an empirical measure of rescattering in the deuteron, the cross section upper limit for the elementary $\gamma n \to K^-\Theta^+$ reaction is estimated to be a factor of 10 higher, {\it i.e.}, $\sim 3$ nb (95% CL).Comment: 5 figures, submitted to PRL, revised for referee comment

Photodisintegration of 4^4He into p+t

The two-body photodisintegration of $^4$He into a proton and a triton has been studied using the CEBAF Large-Acceptance Spectrometer (CLAS) at Jefferson Laboratory. Real photons produced with the Hall-B bremsstrahlung-tagging system in the energy range from 0.35 to 1.55 GeV were incident on a liquid $^4$He target. This is the first measurement of the photodisintegration of $^4$He above 0.4 GeV. The differential cross sections for the $\gamma$$^4$He$\to pt$ reaction have been measured as a function of photon-beam energy and proton-scattering angle, and are compared with the latest model calculations by J.-M. Laget. At 0.6-1.2 GeV, our data are in good agreement only with the calculations that include three-body mechanisms, thus confirming their importance. These results reinforce the conclusion of our previous study of the three-body breakup of $^3$He that demonstrated the great importance of three-body mechanisms in the energy region 0.5-0.8 GeV .Comment: 13 pages submitted in one tgz file containing 2 tex file and 22 postscrip figure

π0\pi^0 photoproduction on the proton for photon energies from 0.675 to 2.875 GeV

Differential cross sections for the reaction $\gamma p \to p \pi^0$ have been measured with the CEBAF Large Acceptance Spectrometer (CLAS) and a tagged photon beam with energies from 0.675 to 2.875 GeV. The results reported here possess greater accuracy in the absolute normalization than previous measurements. They disagree with recent CB-ELSA measurements for the process at forward scattering angles. Agreement with the SAID and MAID fits is found below 1 GeV. The present set of cross sections has been incorporated into the SAID database, and exploratory fits have been extended to 3 GeV. Resonance couplings have been extracted and compared to previous determinations.Comment: 18 pages, 48 figure