Evolution of supergene families associated with insecticide resistance

The emergence of insecticide resistance in the mosquito poses a serious threat to the efficacy of many malaria control programs. We have searched the Anopheles gambiae genome for members of the three major enzyme families- the carboxylesterases, glutathione transferases, and cytochrome P450s-that are primarily responsible for metabolic resistance to insecticides. A comparative genomic analysis with Drosophila melanogaster reveals that a considerable expansion of these supergene families has occurred in the mosquito. Low gene orthology and little chromosomal synteny paradoxically contrast the easily identified orthologous groups of genes presumably seeded by common ancestors. In A. gambiae, the independent expansion of paralogous genes is mainly a consequence of the formation of clusters among locally duplicated genes. These expansions may reflect the functional diversification of supergene families consistent with major differences in the life history and ecology of these organisms. These data provide a basis for identifying the resistance-associated enzymes within these families. This will enable the resistance status of mosquitoes, flies, and possibly other holometabolous insects to be monitored. The analyses also provide the means for identifying previously unknown molecules involved in fundamental biological processes such as development.     

Rapid sedimentological and biological assessment of hydrocarbon contaminated sediments

It is possible to rapidly detect the presence of high concentrations of sediment associated hydrocarbons using a sediment profile camera and simultaneously evaluate the general sedimentological and biological character of a contaminated area. In sediments that were heavily contaminated with hydrocarbons from spills and chronic long-term additions the presence of hydrocarbons was seen about 50% of the time in the sediment profile images as unique features, ‘H spots’. The presence of these features was related to the concentration of hydrocarbons in the sediment. In highly contaminated muddy sediments ‘H spots’ were found in images collected at stations that had from 270 to 610 ppt total hydrocarbons. Sedimentological and biological information obtained from the sediment profile images confirmed the impacted nature of Elizabeth River sediments. Sediment profile imaging provide a means of obtaining an overall evaluation of the quality of a habitat and impacts on that habitat from pollution related environmental disturbances. While qualitative, an advantage of sediment profile image data is that they can be evaluated in less than a day and used to quickly locate inclusions of hydrocarbons in the sediments for further quantitative chemical or biological sampling, or mapping of heavily contaminated areas.     

Chronic vascular catheterisation of the koala and the rabbit

SUMMARY: Vascular access ports were surgically placed, first in rabbits maintained under laboratory conditions, and then in koalas maintained in a wildlife park. The ports remained patent for 9 months in rabbits and for up to 13 months in the koalas and were removed successfully. They allowed collection of blood samples without assistance or disturbance in koalas, and without stress as reflected by plasma cortisol concentration. The use of retention rings on the cannula tubing is recommended.     

Isotope-edited nuclear magnetic resonance: novel methodologies for investigating metabolism

While the use of NMR and stable isotopes in metabolism studies is hardly new, it is only recently that isotope-edited NMR spectroscopy has been applied in kinetic studies of glyphosate metabolism of soil microbes. NMR can detect multiple species simultaneously and non-destructively, yielding valuable information on structural identification of metabolites. T riple R esonance I sotope ED ited spectroscopy (TRIED), [²H]NMR, and [²H–¹³C] INEPT (I nsensitive N ucleus E nhancement through P olarization T ransfer) are three isotope-edited techniques which have been used in combination to examine the microbial degradation of glyphosate (N-phosphonomethylglycine). Using ¹³C- and ¹⁵N-labeled glyphosate, TRIED can detect multiple metabolites in crude matrices at submicrogram levels, an improvement over earlier techniques where milligrams were needed. It can detect 500 nanograms of ¹³C–¹⁵N-labeled compound in a crude sample (1 : 1400 mass ratio), only a few hours work being required. [²H]NMR and [²H–¹³C]INEPT were also used as complementary techniques to further examine metabolites whose ¹³C–¹⁵N bond has been cleaved. The three-isotope-edited methods produced results consistent with both radioactivity and HPLC analyses. Accordingly, we are able to detect minute levels of metabolites in the presence of complex mixtures, minimizing the costs and time of sample purification. ©1999 Society of Chemical Industry     

The effects of duration and concentration of plasma progesterone on the fertility of post-partum cows treated with pregnant mare serum gonadotrophin and intravaginal progesterone

The effects of duration of insertion of progesterone releasing devices (PRIDs) on the concentrations of plasma progesterone (plasma P4) and the relationship of these with fertility were examined respectively in ovariectomised and intact Bos taurus cattle. In experiment 1, ovariectomised cows (n = 14) were treated with PRIDs to produce high and low plasma P4 for 7 or 14 days giving 4 treatment groups designated H7, H14, L7 or L14. Mean plasma P4 in the H14 and L14 groups were 6.6 and 6.0 ng/ml respectively, during the first week and 5.4 and 3.8 ng/ml respectively, during the second. Mean plasma P4 of 6.3 and 4.2 ng/ml were observed in H7 and L7. In experiment 2, suckled cows (n = 171) received the same PRID treatments as cows in experiment 1, plus either 375 or 750 iu pregnant mare serum gonadotrophin, and subsequent calving rates were recorded. These were significantly lower after L7 than the other treatments (12% v 38%, P less than 0.025). These results showed that fertility after oestrus was closely related to plasma P4 observed before oestrus and indicated that treatment with progesterone for as little as 1 week can produce normal fertility providing plasma P4 attain a critical threshold during treatment.     

Effects of suckling on oestradiol benzoate induced LH release in post-partum, ovariectomised cows

Twelve suckling and 12 non-lactating post-partum cows were treated with a progesterone-releasing pessary for 7 or 14 d followed by an injection of 500 micrograms oestradiol benzoate (ODB) 24 h after pessary removal or no injection. Suckling had no significant effect on plasma progesterone concentrations (plasma P4) or plasma luteinising hormone concentrations (plasma LH) during pessary insertion. After pessary removal plasma LH rose in response to ODB. The extent of the rise was similar in suckling and non-lactating cows treated with progesterone for 14 d but was significantly lower in non-lactating than suckling cows (P less than 0.025) treated for 7 d. These findings indicate that suckling increases the responsiveness of plasma LH in post-partum cows following progesterone and oestrogen treatment compared to non-lactating cows and that extended treatment with progesterone can remove this difference.     

The influence of precipitation pulses on soil respiration - Assessing the “Birch effect” by stable carbon isotopes

Sudden pulse-like events of rapidly increasing CO₂-efflux occur in soils under seasonally dry climates in response to rewetting after drought. These occurrences, termed “Birch effect”, can have a marked influence on the ecosystem carbon balance. Current hypotheses indicate that the “Birch” pulse is caused by rapidly increased respiration and mineralization rates in response to changing moisture conditions but the underlying mechanisms are still unclear. Here, we present data from an experimental field study using straight-forward stable isotope methodology to gather new insights into the processes induced by rewetting of dried soils and evaluate current hypotheses for the “Birch“-CO₂-pulse. Two irrigation experiments were conducted on bare soil, root-free soil and intact vegetation during May and August 2005 in a semi-arid Mediterranean holm oak forest in southern Portugal. We continuously monitored CO₂-fluxes along with their isotopic compositions before, during and after the irrigation. δ¹³C signatures of the first CO₂-efflux burst, occurring immediately after rewetting, fit the hypothesis that the “Birch” pulse is caused by the rapid mineralization of either dead microbial biomass or osmoregulatory substances released by soil microorganisms in response to hypo-osmotic stress in order to avoid cell lyses. The response of soil CO₂-efflux to rewetting was smaller under mild (May) than under severe drought (August) and isotopic compositions indicated a larger contribution of anaplerotic carbon uptake with increasing soil desiccation. Both length and severity of drought periods probably play a key role for the microbial response to the rewetting of soils and thus for ecosystem carbon sequestration.     

Animal health monitoring and surveillance in Switzerland

Switzerland has traditionally used a passive disease reporting system for all notifiable diseases. This type of system is not suitable for the documentation of very low prevalences (freedom from disease), sub-clinical cases and non-notifiable diseases. In order to meet the high international standards for animal health surveillance and to fulfil the general need for sound animal health data, Switzerland has evaluated the feasibility of modern monitoring and surveillance concepts. In general, the principle of active surveillance has been acquired and is now being applied whenever possible. In this paper, several examples of Swiss surveillance systems are presented and discussed. They include systematic testing of random population samples, carcase screening at abattoirs and sentinel herd monitoring.