Improved isolation procedure for azaspiracids from shellfish, structural elucidation of azaspiracid-6, and stability studies

Azaspiracids are a group of lipophilic polyether toxins produced by the small dinoflagellate Azadinium spinosum. They may accumulate in shellfish and can result in illnesses when consumed by humans. Research into analytical methods, chemistry, metabolism, and toxicology of azaspiracids has been severely constrained by the scarcity of high-purity azaspiracids. Consequently, since their discovery in 1995, considerable efforts have been made to develop methods for the isolation of azaspiracids in sufficient amounts and purities for toxicological studies, in addition to the preparation of standard reference materials. A seven-step procedure was improved for the isolation of azaspiracids-1-3 (1, 2, and 3) increasing recoveries 2-fold as compared to previous methods and leading to isolation of sufficiently purified azaspiracid-6 (6) for structural determination by NMR spectroscopy. The procedure, which involved a series of partitioning and column chromatography steps, was performed on 500 g of Mytilus edulis hepatopancreas tissue containing ~14 mg of 1. Overall yields of 1 (52%), 2 (43%), 3 (43%), and 6 (38%) were good, and purities were confirmed by NMR spectroscopy. The structure of 6 was determined by one- and two-dimensional NMR spectroscopy and mass spectrometry. The stability of 6 relative to 1 was also assessed in three solvents in a short-term study that demonstrated the greatest stability in aqueous acetonitrile.     

Gaseous and leaching nitrogen losses from no-tillage and conventional tillage systems following surface application of cattle manure

Previous studies have demonstrated inconsistent results on the impact of tillage systems on nitrogen (N) losses from field-applied manure. This study assessed the impact of no-tillage (NT) and conventional tillage (CT) systems on gaseous N losses, N₂O:N₂O + N₂ ratios and NO₃⁻-N leaching following surface application of cattle manure. The study was undertaken during the 2003/2004 and 2004/2005 seasons at two field sites in Nova Scotia namely, Streets Ridge (SR) in Cumberland County and the Bio-environmental Engineering Centre (BEEC) in Truro. Results showed that the NT system had higher (p < 0.05) NH₃ losses than CT. Over the two seasons, manure incorporation in CT reduced NH₃ losses on average by 86% at SR and 78% at BEEC relative to NT. At both sites and during both seasons, denitrification rates and N₂O fluxes in NT were generally higher than in CT plots, presumably due to higher soil water and organic matter content in NT. Over the two seasons, mean denitrification rates at SR were 239 and 119 g N ha⁻¹ d⁻¹, while N₂O fluxes were 120 and 64 g N ha⁻¹ d⁻¹ under NT and CT, respectively. At BEEC mean denitrification rates were 114 and 71 g N ha⁻¹ d⁻¹, while N₂O fluxes were 52 and 27 g N ha⁻¹ d⁻¹ under NT and CT, respectively. Conversely, N₂O:N₂O + N₂ ratios were lower in NT than CT suggesting more complete reduction of N₂O to N₂ under NT. When averaged across all soil depths, NO₃⁻-N was higher (p < 0.05) in CT than NT. Nitrate-N decreased with depth at both sites regardless of tillage. In most cases, NO₃⁻-N was higher under CT than NT at all soil depths. Similarly, flow-weighted average NO₃⁻-N concentrations in drainage water were generally higher under CT. This may be partly attributed to higher denitrification rates under NT. Therefore, NT may be a viable strategy to remove NO₃⁻-N from the soil, and thus, reduce NO₃⁻-N contamination of groundwater. However, it should be noted that while the use of NT reduces NO₃⁻-N leaching it may come with unintended environmental tradeoffs, including increased NH₃ and N₂O emissions.     

Soil microbial biomass, activity and community composition in adjacent native and plantation forests of subtropical Australia

Purpose

Soil nitrogen (N) availability is a critical determinant of plantation productivity in subtropical Australia and is influenced by the soil microbial community. The size, structure and function of the soil microbial community can be impacted by land-use change and residue management. The objectives of this study were to examine the impact of land-use change from (1) native forest (NF) to first rotation (1R) hoop pine plantation and (2) 1R hoop pine plantation to second rotation (2R) hoop pine plantation on the soil microbial community. The impact of residue management on the soil microbial community was also investigated in the 2R forest, where soil microbial parameters were measured in tree rows (2R-T) and windrows (2R-W). In addition, relationships between soil microbial parameters and soil N parameters were investigated.

Materials and methods

Each of the four treatments (NF, 1R, 2R-T and 2R-W) had five 24-m² replicate plots from which 15 soil cores were collected and bulked at three depths (0–10, 10–20, 20–30 cm). Microbial biomass carbon (MBC) and N (MBN) and soil respiration were measured on field moist soils. In addition, carbon (C) source utilisation patterns were assessed using the whole soil MicroResp? technique (Campbell et al. 2003).

Results and discussion

Results indicate that the land-use change from NF to 1R hoop pine plantation significantly reduced MBC, respiration rate, soil total C and total N. Furthermore, the land-use change appeared to have a significant impact on the soil microbial community composition measured using MicroResp? profiles. Land-use change from 1R to 2R hoop pine plantation resulted in a decline in total C and MBN and a shift in microbial community composition. When compared to the 2R-T soils, the 2R-W soils tended to have a greater microbial biomass and respiration rate. Residue management also influenced the microbial community composition measured in the MicroResp? profiles.

Conclusions

Results indicate that land-use change had a significant impact on the soil microbial community, which was likely to be related to shifts in the quality and quantity of organic inputs associated with the change in land use. This may have significant implications for the long-term productivity of the soil resource. Further studies are required to confirm a difference in microbial community composition associated with residue management. In addition, long-term experiments in subtropical Australia are necessary to verify the results of this snapshot study and to improve our understanding of the impact of single-species plantation forestry and residue management on the soil microbial community, soil N dynamics and ultimately the long-term sustainability of the soil resource.     

Ectopic hepatocellular carcinoma in a dog

CASE HISTORY: A 14-year-old neutered male Bearded Collie was presented with a history of recurrent, intermittent urinary incontinence of 7 years duration. CLINICAL FINDINGS: A large, firm, non-painful mass was found in the mid-abdominal region on palpation. Ultrasonography of the mass revealed a compartmentalised structure with mixed echogenicity, and which did not appear to be associated with any of the abdominal organs. Ultrasound-guided fine needle aspirates contained several clusters of epithelial cells with cytological features of hepatocytes. At exploratory laparotomy, the mass was found in the gastrosplenic ligament within the greater omentum. PATHOLOGICAL FINDINGS AND DIAGNOSIS: Histopathologically, the mass consisted of sheets of hepatocytes, but without the characteristic hepatic architecture. The cells showed moderate variation in nuclear size and were sometimes binucleate. A diagnosis of hepatocellular carcinoma (HCC) in the mesentery was made. CLINICAL RELEVANCE: The presence of ectopic hepatic tissue has been reported rarely in man and cats, but not in the dog. Neoplastic transformation of ectopic hepatic tissue is seen in man. This is the first report of the presentation, clinical findings and treatment of a dog with ectopic HCC.     

影响烟草特有亚硝胺积累的因素

把Ｎｉｃｏｔｉａｎａ ｐｌｕｍｂａｇｉｎｉｆｏｌｉａ的基因突变种子和普通种子发芽后嫁接在Ｗｅｓｃｏｎｓｉｎ３８烟株上,与温室条件下正常盆栽生长的Ｎｉｃｏｔｉａｎａ ｐｌｕｍｂａｇｉｎｉｆｏｌｉａ和母体Ｗｅｓｃｏｎｓｉｎ３８分析比较,试验结果得出：Ｗｅｓｃｏｎｓｉｎ３８上嫁接Ｎｉｃｏｔｉａｎａ ｐｌｕｍｂａｇｉｎｉｆｏｌｉａ基因突变种子还原酶活性最高,晾制后烟叶中烟草特有的亚硝胺含量次之;温室盆栽的     

Recurrent flank alopecia in a Tibetan Terrier

Recurrent flank alopecia is described in a 2-year-old, male, neutered Tibetan Terrier with concurrent atopic dermatitis. The diagnosis of recurrent flank alopecia was made after 3 consecutive years of localised, winter-onset alopecia. The diagnosis was based on history, compatible clinical signs and supportive histopathology. The diagnosis was complicated by the presence of concurrent pruritic dermatitis. To our knowledge this is the first report of recurrent flank alopecia in this breed.     

Identification of a dynamic atmosphere at Enceladus with the Cassini magnetometer

The Cassini magnetometer has detected the interaction of the magnetospheric plasma of Saturn with an atmospheric plume at the icy moon Enceladus. This unanticipated finding, made on a distant flyby, was subsequently confirmed during two follow-on flybys, one very close to Enceladus. The magnetometer data are consistent with local outgassing activity via a plume from the surface of the moon near its south pole, as confirmed by other Cassini instruments.