Continuous in vitro culture of the carpet shell clam Tapes decussatus protozoan parasite Perkinsus atlanticus

Continuous in vitro cultures of the clam Tapes decussatus parasite Perkinsus atlanticus were established from infected gill fragments, infected haemolymph and parasite hypnospores isolated from infected gill fragments following incubation in Ray's fluid thioglycollate medium (RFTM). No continuous cultures could be initiated from P. atlanticus zoospores. Cultures initiated from hypnospores yielded the highest percentage of continuous cultures (100%, 6/6), followed by cultures initiated from gill fragments (93%, 43/46) and from haemolymph (30%, 3/10). Failures to establish continuous cultures were due to microbial contamination. The source of parasite influenced the success rate, the time taken to establish cultures and the size of cultured cells. In vitro proliferation of parasite cells was mainly by vegetative multiplication. Zoosporulation, yielding motile biflagellated zoospores, was observed at a low frequency (< 1% of dividing cells) in every culture. Morphology of cultured cells examined with light and transmission electron microscopy corresponded to that of P. atlanticus found in clam tissues. Cultured cells enlarged in RFTM and stained blue-black with Lugol's solution, which are characteristics of the Perkinsus species cells. DNA sequences of the internal transcribed spacer (ITS) region of the ribosomal RNA gene complex matched those of P. atlanticus. All cultures were established in a medium designated JL-ODRP-2A that was similar in composition to the culture medium JL-ODRP-1 originally used to propagate Perkinsus marinus in vitro. Proliferation of P. atlanticus in vitro could be supported by the commercial culture medium (1:2 v/v) DME:Ham's F-12 with fetuin.     

Alfalfa weevil (Coleoptera: Curculionidae) larval sampling: comparison of shake-bucket and sweep-net methods and effect of training

Two field-sampling methods, shake-bucket and sweep-net, were compared for use in monitoring alfalfa weevil, Hypera postica (Gyllenhal), larvae in alfalfa hay of the high plains and intermountain region of North America. In this region, alfalfa grows to sufficient height to use both methods before the more damaging late instars peak in abundance. Both methods also were compared with extracting larvae by using Berlese funnels in the laboratory. The shake-bucket method was more sensitive in detecting small larvae (first and second instars) than large larvae (third and fourth instars), and the sweep-net method detected a lower proportion of small larvae. The number of larvae collected with the shake-bucket method was strongly correlated with number of larvae recovered from Berlese funnels (total larval counts, R2 = 0.85). Correlation of the sweep-net samples with the Berlese extraction was also significant but less strong (R2 = 0.56). In addition, sampler instruction was evaluated to determine whether demonstration training affects performance of inexperienced samplers using the two field-sampling methods. Training did not significantly change sampler performance in using the shake-bucket but did increase the number of large larvae when using the sweep-net. In addition, less variability was associated with the shake-bucket sampling method than with the sweep-net method for samplers who only had access to written sampling instructions. Therefore, when estimation of small larval abundance is desired for economic decision-making and sampling is performed by people with little or infrequent sampling experience, such as growers, the shake-bucket method is the preferred field-sampling method in the high plains and intermountain region of North America Sweep-net sampling is more variable than shake-bucket sampling, although demonstration training improves the usefulness of the sweep-net.     

Diversity of cuticular wax among Salix species and Populus species hybrids

The leaf cuticular waxes of three Salix species and two Populus species hybrids, selected for their ability to produce high amounts of biomass, were characterized. Samples were extracted in CH(2)Cl(2) three times over the growing season. Low kV SEM was utilized to observe differences in the ultrastructure of leaf surfaces from each clone. Homologous series of wax components were classified into organic groups, and the variation in wax components due to clone, sample time, and their interaction was identified. All Salix species and Populus species hybrids showed differences in total wax load at each sampling period, whereas the pattern of wax deposition over time differed only between the Salix species. A strong positive relationship was identified between the entire homologous series of alcohols and total wax load in all clones. Similarly strong relationships were observed between fatty acids and total wax load as well as fatty acids and alcohols in two Salix species and one Populus species hybrid. One Salix species, S. dasyclados, also displayed a strong positive relationship between alcohols and alkanes. These data indicate that species grown under the same environmental conditions produce measurably different cuticular waxes and that regulation of wax production appears to be different in each species. The important roles cuticular waxes play in drought tolerance, pest, and pathogen resistance, as well as the ease of wax extraction and analysis, strongly suggest that the characteristics of the cuticular wax may prove to be useful selectable traits in a breeding program.     

Processes that occur before second cleavage determine third cleavage orientation in Xenopus

As in many organisms, the first three cleavage planes of Xenopus laevis eggs form in a well-described mutually orthogonal geometry. The factors dictating this simple pattern have not been unambiguously identified. Here, we describe experiments, using static magnetic fields as a novel approach to perturb normal cleavage geometry, that provide new insight into these factors. We show that a magnetic field applied during either or both of the first two cell cycles can induce the third cell cycle mitotic apparatus (MA) at metaphase and the third cleavage plane to align nearly perpendicular to their nominal orientations without changing cell shape. These results indicate that processes occurring during the first two cell cycles primarily dictate the third cleavage plane and mitotic apparatus orientation. We discuss how mechanisms that can align the MA after it has formed are likely to be of secondary importance in determining cleavage geometry in this system.     

Elemental and organochlorine residues in bald eagles from Adak Island,Alaska

Adak Island is a remote island in the Aleutian Island archipelago of Alaska (USA) and home to various military activities since World War II. To assess the contaminant burden of one of Adak Island's top predators, livers and kidneys were collected from 26 bald eagle (Haliaeetus leucocephalus) carcasses between 1993 and 1998 for elemental and organochlorine analyses. Mean cadmium, chromium, mercury, and selenium concentrations were consistent with levels observed in other avian studies and were below toxic thresholds. However, elevated concentrations of chromium and mercury in some individuals may warrant concern. Furthermore, although mean polychlorinated biphenyl and pp'-dichlorodiphenyldichloroethylene concentrations were below acute toxic thresholds, they were surprisingly high given Adak Island's remote location.     

Applications of Tree-Maps to hierarchical biological data

SUMMARY: A brief overview of Tree-Maps provides the basis for understanding two new implementations of Tree-Map methods. TreeMapClusterView provides a new way to view microarray gene expression data, and GenePlacer provides a view of gene ontology annotation data. We also discuss the benefits of Tree-Maps to visualize complex hierarchies in functional genomics. AVAILABILITY: Java class files are freely available at http://mendel.mc.duke.edu/bioinformatics/ CONTACT: mccon012@mc.duke.edu SUPPLEMENTARY INFORMATION: For more information on TreeMapClusterView (see http://mendel.mc.duke.edu/bioinformatics/software/boxclusterview/), and http://mendel.mc.duke.edu/bioinformatics/software/geneplacer/).     

Pulmonary carbonic anhydrase in the garter snake, Thamnophis sirtalis

This study examined whether the snake lung possesses intravascular carbonic anhydrase (CA). Lungs were perfused with control salines and with salines containing CA inhibitors. Perfusion with control salines resulted in a stable CO(2) excretion, whereas CA inhibitors significantly reduced pulmonary CO(2) excretion. Membrane-permeable and membrane-impermeable CA inhibitors produced comparable decreases in CO(2) excretion, suggesting that extracellular, intravascular CA participated in the pulmonary CO(2)-HCO3(-)-H(+) reactions. Treatment of lungs with phosphatidylinositol specific-phospholipase C (PI-PLC) significantly decreased CO(2) excretion, indicating that CA was connected to the luminal endothelial cell membrane by a phosphatidylinositol glycan linkage. Taken together, these results are the first to demonstrate the presence of membrane-bound, intravascular CA (CA IV) in the snake lung.