A novel method for the transport and analysis of genetic material from polyps and zooxanthellae of scleractinian corals

We have developed a new simple method for transport, storage, and analysis of genetic material from the corals Agaricia agaricites, Dendrogyra cylindrica, Eusmilia ancora, Meandrina meandrites, Montastrea annularis, Porites astreoides, Porites furcata, Porites porites, and Siderastrea siderea at room temperature. All species yielded sufficient DNA from a single FTA(R) card (19 mug-43 ng) for subsequent PCR amplification of both coral and zooxanthellar DNA. The D1 and D2 variable region of the large Subunit rRNA gene (LSUrDNA) was amplified from the DNA of P. furcata and S. siderea by PCR. Electrophoresis yielded two major DNA bands: an 800-base pair (bp) DNA, which represented the coral ribosomal RNA (rRNA) gene, and a 600-bp DNA, which represented the zooxanthellar srRNA gene. Extraction of DNA from the bands yielded between 290 mug total DNA (S. siderea coral DNA) and 9 mug total DNA (P. furcata zooxanthellar DNA). The ability to transport and store genetic material from scleractinian corals without resort to laboratory facilities in the field allows for the molecular Study of a far wider range and variety of coral sites than have been studied to date. (C) 2003 Elsevier Science B.V. All rights reserved.

Analysis of 16S rDNA sequences from pathogenic Leptospira serovars and use of single nucleotide polymorphisms for rapid speciation by D-HPLC

Leptospira have a worldwide distribution and include important zoonotic pathogens yet diagnosis and differentiation still tend to rely on traditional bacteriological and serological approaches. In this study a 1.3 kb fragment of the rrs gene (16S rDNA) was sequenced from a panel of 22 control strains, representing serovars within the pathogenic species Leptospira interrogans, Leptospira borgpetersenii, and Leptospira kirschneri, to identify single nucleotide polymorphisms (SNPs). These were identified in the 5' variable region of the 16S sequence and a 181 bp PCR fragment encompassing this region was used for speciation by Denaturing High Performance Liquid Chromatography (D-HPLC). This method was applied to eleven additional species, representing pathogenic, non-pathogenic and intermediate species and was demonstrated to rapidly differentiate all but 2 of the non-pathogenic Leptospira species. The method was applied successfully to infected tissues from field samples proving its value for diagnosing leptospiral infections found in animals in the UK. Crown Copyright (C) 2010 Published by Elsevier Ltd. All rights reserved.

Trace element geochemistry from the Birrimian metasediments of the Northern Region of Ghana

Trace element distributions in rock, soil and groundwater from of the Birrimian metasediments and granites located in the Northern Region of Ghana are described. High positive correlations are observed between selected major elements and trace metals (e.g. K2O and Rb, Al2O3 and V, Fe2O3 and V, and K2O and Y) in rocks and soils, and attributed to the presence of major source minerals. Ca and Sr were strongly correlated in groundwater, suggesting greater water-rock interaction. Low association of V with Fe is explained by (i) relatively higher mobility of V as against Fe; (ii) low Fe content in the parent rocks and (iii) variable sources of Fe and V.

In situ degradability of dry matter and neutral-detergent fibre of thorn scrubland forage consumed by goats in the semi-arid region of north Mexico

Three goats provided with oesophageal and ruminal cannulae were used to determine variations in dry matter (DM) and neutral-detergent fibre (NDF) degradability of the forage consumed when grazing thorn scrubland in the semi-arid region of north Mexico, during two consecutive dry and wet periods. Ingesta samples were incubated intraruminally, the data were fitted to the exponential equation P = a + b (1-e(-ct)) and statistically analysed using a randomized-block design. Organic matter and crude protein (CP) contents were higher (P < 0.05) in the wet seasons. Values of NDF were similar in dry and wet season of both years whereas higher numerical values of acid-detergent fibre (ADF), lignin and cellulose were registered in the dry seasons. DM and NDF degradabilities after 24 and 48 h of ruminal incubation were higher (P < 0.05) in the wet seasons. Higher values (P < 0.05) in DM and NDF bag losses at zero time (A fraction) were registered in the two wet seasons. The insoluble but fermentable DM and NDF (B fractions) were higher (P < 0.05) in the 1999 wet season and variable in the rest of the studied period. Numerically higher values of DM and NDF c fraction were found in wet periods, whereas DM and NDF potential degradabilities were higher (P < 0.05) in the wet season in 1999 and similar across seasons in 2000. Lowest (P < 0.05) contents of CP in grazed forage, DM and NDF degradabilities after 48 h of ruminal incubation, and A, and B, and c fractions were observed in the dry seasons. Thus, these results may be related to both the lower feeding value of forage consumed by the animals and lower performance of livestock during this period. Then, the DM and NDF degradability after 48 h, together with the insoluble but fermentable matter and the c fraction permit the nutritive value of the forage consumed by grazing goats to be accurately described.

Characterization of the 23S and 5S rRNA genes and 23S-5S intergenic spacer region (ITS-2) of Photobacterium damselae

The 23S ribosomal RNA (rRNA) gene has been sequenced in strains of the fish pathogens Photobacterium damselae subsp. damselae (ATCC 33539) and subsp. piscicida (ATCC 29690), showing that 3 nucleotide positions are clearly different between subspecies. In addition, the 5S rRNA gene plus the intergenic spacer region between the 23S and 5S rRNA genes (ITS-2) were amplified, cloned and sequenced for the 2 reference strains as well as the field isolates RG91 (subsp. damselae) and DI21 (subsp. piscicida). A 100% similarity was found for the consensus 5S rRNA gene sequence in the 2 subspecies, although some microheterogeneity was detected as inter-cistronic variability within the same chromosome. Sequence analysis of the spacer region between the 23S and 5S rRNA genes revealed 2 conserved and 3 variable nucleotide sequence blocks, and 4 different modular organizations were found. The ITS-2 spacer region exhibited both inter-subspecies and inter-cistronic polymorphism, with a mosaic-like structure. The EMBL accession numbers for the 23S, 5S and ITS-2 sequences are: P. damselae subsp. piscicida 5S gene (AJ274379), P. damselae subsp. damselae 23S gene (Y18520), subsp. piscicida 23S gene (Y17901), R damselae subsp. piscicida ITS-2 (AJ250695, AJ250696), P. damselae subsp. damselae ITS-2 (AJ250697, AJ250698).

The neural bases of the short-term storage of verbal information are anatomically variable across individuals

What are the precise brain regions supporting the short-term retention of verbal information? A previous functional magnetic resonance imaging (fMRI) study suggested that they may be topographically variable across individuals, occurring, in most, in regions posterior to prefrontal cortex (PFC), and that detection of these regions may be best suited to a single-subject (SS) approach to fMRI analysis (Feredoes and Postle, 2007). In contrast, other studies using spatially normalized group-averaged (SNGA) analyses have localized storage-related activity to PFC. To evaluate the necessity of the regions identified by these two methods, we applied repetitive transcranial magnetic stimulation (rTMS) to SS- and SNGA-identified regions throughout the retention period of a delayed letter-recognition task. Results indicated that rTMS targeting SS analysis-identified regions of left perisylvian and sensorimotor cortex impaired performance, whereas rTMS targeting the SNGA-identified region of left caudal PFC had no effect on performance. Our results support the view that the short-term retention of verbal information can be supported by regions associated with acoustic, lexical, phonological, and speech-based representation of information. They also suggest that the brain bases of some cognitive functions may be better detected by SS than by SNGA approaches to fMRI data analysis.