Balamuthia mandrillaris therapeutic mud bath in Jamaica.

Balamuthia mandrillaris is an emerging cause of encephalitis in humans. The transmission dynamics are poorly understood due to the high fatality rate and the sporadic nature of cases. Seventy-two soil samples were collected from beaches and the banks of lagoons, rivers, ponds, mineral springs and streams from across Jamaica and assayed for the presence of B. mandrillaris. Seventy-nine sites were sampled and the mitochondrial 16S rDNA gene of B. mandrillaris was amplified and sequenced to confirm the presence of the amoeba. One isolate of B. mandrillaris was recovered from soil from mineral spring which hosts an informal therapeutic mud bath business. Although B. mandrillaris is less frequently isolated from soil than other free-living amoebae, rubbing mud containing the organism onto the skin increases the likelihood of exposure and infection. This first report on the isolation of B. mandrillaris in the Caribbean and its presence in soil where human contact is likely warrants further investigation using serological methods to elucidate exposure patterns.

Comparison of secondary ion mass spectrometry and micromilling/continuous flow isotope ratio mass spectrometry techniques used to acquire intra-otolith delta18O values of wild Atlantic salmon (Salmo salar).

The chemical signals in the sequential layers of fish otoliths have the potential to provide fisheries biologists with temporal and spatial details of migration which are difficult to obtain without expensive tracking methods. Signal resolution depends, however, on the extraction technique used. We compared the use of mechanical micromilling and continuous flow isotope ratio mass spectrometry (CF-IRMS) methods with secondary ion mass spectrometry (SIMS) to obtain delta(18)O profiles from otoliths of wild Atlantic salmon (Salmo salar) and used these to corroborate the time of freshwater emigration of the juvenile with macroscopic patterns within the otolith. Both techniques showed the transition occurring at the same visible feature on the otolith, allowing future analyses to easily identify the juvenile (freshwater) versus adult (marine) life-stages. However, SIMS showed a rapid and abrupt transition whereas micromilling provided a less distinct signal. The number of samples that could be obtained per unit area sampled using SIMS was 2 to 3 times greater than that when using micromilling/CF-IRMS although the delta(18)O values and analytical precisions (approximately 0.2 per thousand) of the two methods were comparable. In addition, SIMS delta(18)O results were used to compare otolith aragonite values with predicted values calculated using various isotope fractionation equations.

Characterization of Melanoplus sanguinipes oviposition stimulating protein expression and re-examination of its potential role in stimulating oviposition.

Melanoplussanguinipes oviposition stimulating protein (MsOSP) was characterized and its role in stimulating oviposition in virgin females was examined. A 967nt MsOSP mRNA sequence with homology to previously characterized N-terminal amino acid sequence data for MsOSP was identified in a RNAseq library generated from an mRNA pool from the long hyaline tubule (LHT) of the male accessory gland complex. This transcript contained a predicted 729nt open reading frame encoding the 242aa putative MsOSP protein and had the second highest read abundance in the library. The MsOSP transcript was detected exclusively in the LHT tissue of adult males and its abundance increased with time until 7 days post-eclosion. Western blot analysis using an anti-MsOSP antibody showed high levels of MsOSP protein in the LHT luminal secretions of virgin males and to a lesser degree was associated with the aedeagus and ejaculatory duct. MsOSP was shown to be a major protein component of the spermatophore packet transferred from the male to female during copulation. However, only minor amounts of MsOSP could be detected in the female bursa, spermatheca and oviduct. Intrahemocoelic injection of LHT luminal protein into mature virgin females stimulated oviposition in ∼ 65% of females. A similar but non-significant trend was observed upon injection of purified recombinant MsOSP protein, and immunoprecipitation of LHT protein with anti-MsOSP antibody led to abrogation of oviposition stimulation upon injection of mature virgin females. Despite the demonstration of stimulation of oviposition upon intrahemocoelic injection of LHT-derived-MsOSP into mature virgin females, the potential mode of action of MsOSP in this process remains to be determined. MsOSP cannot be detected in the tissues other than the bursa, spermatheca and oviduct of female grasshoppers and relatively large quantities of MsOSP are required to stimulate oviposition upon injection.

Quantitative classification of adhesive bondline dimensions using Lamb waves and artificial neural networks.

Adhesive bonding of metal assemblies is gaining acceptance for use with safety critical structures, and there is a need for effective inspection for both quality assurance (QA) and the assessment of condition in service. One aspect of QA is the need for the dimensions of adhesive bondlines to be within tolerance and measurable. This paper describes the application of ultrasonic Lamb waves in the determination of the principal dimensions of two forms of adhered joints (Lap and T-form) between metal plates. Low order Lamb wave modes (s0 and a1) are propagated across adhered bond-lines, and the received signals are transformed to the modulus frequency domain (FD). The FD data are used as input to artificial neural networks (ANNs), which are trained to associate features in the input data with principal bondline dimensions. The performance of different network structures and simplified forms of these is examined, and the technique gives reliable estimates of the required dimensions in bondlines not included in network training. The interconnected weights of simplified networks provide evidence of the features in Lamb wave signals that underlie the successful operation of the method.

Seasonal variation in the effects of hard substratum biofilming on settlement of marine invertebrate larvae.

The effect of season on "biofilming";, as a cue for the settlement of marine invertebrate larvae, was investigated in a long-term field study during the years 1992-1994. The series of settlement experiments was conducted in a tidal rapid on the west coast of Scotland, and involved manipulations of artificial panels. Biofilming of substrata, whilst excluding larval settlement, was achieved by the enclosure of panels within tight-fitting (but removable) mesh screens so that the number of settlers on filmed and unfilmed substrata were counted in the initial absence of other incumbent post-larvae. Depending on larval species, the effects of biofilming were found to be either facilitatory or inhibitory. Significant within- and between-species seasonal differences in the settlement responses were detected, and a reversal of the effect of biofilming on larval settlement response, from inhibitory to facilitatory and vice versa, was noted with season in the case of some taxonomic groups and species (e.g. Tubulipora sp., Plagioecia sp., Electra pilosa (L.)). The present study emphasizes the need for extended field studies of larval responses to environmental cues, when the focus of interest is in drawing general inferences about naturally occurring behavioural patterns at settlement.

Regulation of loblolly pine (Pinus taeda L.) arginase in developing seedling tissue during germination and post-germinative growth.

After seed germination, hydrolysis of storage proteins provides a nitrogen source for the developing seedling. In conifers the majority of these reserves are located in the living haploid megagametophyte tissue. In the developing loblolly pine (Pinus taeda L.) seedling an influx of free amino acids from the megagametophyte accompanies germination and early seedling growth. The major component of this amino acid pool is arginine, which is transported rapidly and efficiently to the seedling without prior conversion. This arginine accounts for nearly half of the total nitrogen entering the cotyledons and is likely a defining factor in early seedling nitrogen metabolism. In the seedling, the enzyme arginase is responsible for liberating nitrogen, in the form of ornithine and urea, from free arginine supplied by the megagametophyte. In this report we investigate how the seedling uses arginase to cope with the large arginine influx. As part of this work we have cloned an arginase cDNA from a loblolly pine expression library. Analysis of enzyme activity data, accumulation of arginase protein and mRNA abundance indicates that increased arginase activity after seed germination is due to de novo synthesis of the enzyme. Our results suggest that arginase is primarily regulated at the RNA level during loblolly pine seed germination and post-germinative growth.