Microtubule assembly and disassembly at alkaline pH

Although it is now apparent that the intracellular pH may rise considerably above neutrality under physiological conditions, information on the effect of alkaline pH on microtubule assembly and disassembly is still quite fragmentay. We have studied the assembly/disassembly of bovine brain microtubule protein at alkaline pH in vitro. When microtubules are assembled to a new steady state at pH less than 7 and pH is then made more alkaline, they undergo a rapid disassembly to a new steady state. This disassembly is reversed by acidification. The degree of disassembly is determined largely by the pH- dependence of the critical concentration, which increases five to eight times, from pH 7 to 8. A fraction of assembly-incompetent tubulin is identified that increases with pH, but its incompetency is largely reversed with acidification. Measurements of microtubule lengths are used to indicate that disassembly occurs by uniform shortening of microtubules. A comparison of shortening by alkalinization with dilution suggests that the intrinsic rate of disassembly is accelerated by increasing pH. The capacity for initiating assembly is progressively lost with incubation at alkaline pH (although some protection is afforded by sulfhydryl-reducing agents). However, direct assembly from depolymerized mixtures is possible at least up to pH 8.3, and the steady state achieved at these alkaline pH values is stable. Such preparations are readily disassembled by cold and podophyllotoxin (PLN). Disassembly induced by PLN is also markedly enhanced at alkaline pH, suggesting a corresponding enhancement of “treadmilling.” The implications of physiological events leading to alkaline shifts of pH for microtubule assembly/disassembly are discussed, particularly in the light of recent hypotheses regarding treadmilling and its role in controlling the distribution of microtubules in vivo.     

Observations on the biology of Subcoccinella vigintiquattuor-punctata (L.) in southern England

• 1 Subcoccinella 24-punctata (L.) is exclusively phytophagous, and is widely distributed throughout Britain.
• 2 Under laboratory conditions the duration of pre-adult instars is about 6 weeks. Adults appear from the end of June onwards. Many enter reproductive diapause at the end of July, but remain active till the end of October.
• 3 Reproductive diapause is determined by the interaction of photoperiod and temperature.
• 4 Hibernation is unstable, and there may be high mortality in overwintering adults. These finally die during May and June.
• 5 The cool temperate climate limits the population size, preventing it from reaching pest proportions.

     

Catchment and reach-scale properties as indicators of macroinvertebrate species traits

1. We used catchment and reach-scale physical properties to predict the occurrence of specific species life history and behaviour traits of aquatic insects across fifty-eight catchments in a mixed land use basin. Catchment-scale attributes were derived using a geographical information system (GIS). Logistic regression techniques were used to model the relationships.
2. The reach-scale properties were highly predictive of species traits. Fourteen of the fifteen traits had significant models with concordance values greater than 68%. Cross-sectional area at bank full discharge, % shallow, slow-water habitats, and % fines were the most important variables.
3. Life history and behavioural attributes were best related to reach-scale physical features. This suggests that species traits exhibit strong relationships to local environmental conditions.
4. Catchment-scale variables had fewer significant models with species traits (four of fifteen), however these variables may have direct or indirect influence on reach-scale properties.
5. Catchment features, in particular surficial geology, influence macroinvertebrate assemblages through their control over channel morphology and hydrologic patterns.
6. The effects of land use were masked by geology (i.e. lacustrine clay geology and rowcrop agriculture were correlated), lack of detail in land use data and the aggregation of the species data.
7. These models reflect the coupling of local environmental conditions and the set of adaptations among the local taxa. These observations underscore the idea that habitat plays a major role in organizing stream assemblages.
8. Using these approaches, predictions can be made about the ability of various taxonomic groupings to track environmental change through time, or for projecting the impact of alternative land management scenarios. Identifying fundamental life history and other traits can improve the selection and evaluation of such indicators.     

Two New Species of Hartmannella Amebae Infecting Freshwater Mollusks

SYNOPSIS. Two new species of small amebae, Hartmannella biparia n. sp. and Hartmannella quadriparia n. sp., were 1st observed in the freshwater mollusks Bulinus globosus and Biomphalaria pallida, respectively. The amebae multiplied in cytoplasmic vacuoles in host cells, particularly in foci in the mantle collar, foot, and intestinal wall. Both amebae had functional contractile vacuoles while within host cells. H. biparia emerged from intracytoplasmic vacuoles in pairs, H. quadriparia in fours, suggesting characteristic reproductive stages.
H. biparia had limax-type motility by smooth lobopodia, was 22 μ long; with vesicular nucleus 3 μ and central endosome 1.5 μ, multiplied by binary fission and formed spherical, smooth-walled cysts 11 μ in diameter. H. quadriparia had limax-type motility by lobopodia with fine, acute projections; was 10 μ long, with vesicular nucleus 2 μ and central endosome 0.75 μ, multiplied and formed spherical, smooth-walled cysts 5 μ in diameter. Neither species multiplied in cysts outside the host.
H. biparia was found infecting 12 species and experimentally transferred to 4 more species of freshwater snails; H. quadriparia was found in one species and experimentally transferred to 6 other species.     

Evaluation of measurement uncertainties of an analytical method for the determination of aflatoxin M<Subscript>1</Subscript> in milk and milk powder

The mycotoxin aflatoxin M₁ (AfM₁) is a serious food safety hazard for which the European Commission has already established a maximum permissible level of 0.05 μg/kg AfM₁ in milk and products thereof. For control analysis laboratories are increasingly asked to submit full uncertainties of their analytical results.The evaluation of measurement uncertainties of an analytical method for the determination of AfM₁ in milk and milk powder on the basis of ‘in-house’ validation data in compliance with the ‘Guide to the Expression of Uncertainty in Measurement (GUM)’ [1] and the ‘EURACHEM Guide’ [2] is described. A similar approach will be used to assess the performance of methods employed by laboratories participating in the certification of reference materials for AfM₁ in milk powder.     

An invasive crayfish affects egg survival and the potential recovery of an endangered population of Arctic charr

1. Many fish stocks have declined, because of overharvesting, habitat destruction and introduced species. Despite efforts to rehabilitate some of these stocks, not all are responding or are recovering only slowly. 2. In freshwater systems, introduced crayfish are often problematic, and it has been suggested that their egg predation could reduce recruitment in depleted stocks of native fish. 3. Here, we report the results of a field experiment, using experimental cages, on the extent of predation on eggs of great Arctic charr (Salvelinus umbla) in Lake Vättern, Europe’s fifth largest lake. Here, the great Arctic charr has declined dramatically and is listed as critically endangered. 4. We were able to partition the total loss rate of eggs into background mortality, predation by introduced signal crayfish (Pacifastacus leniusculus) and predation by native fish. The mortality rate of charr eggs because of crayfish was estimated at more than five times that because of native fish. Of the total loss of eggs, 80% is believed to be caused by crayfish and 14% by fish, with 6% being natural background mortality. 5. In a worst case scenario, our data infer that only 25% of the original number of eggs would survive, compared with 75% in the absence of crayfish. This could impair recovery of the stock of the endangered great Arctic charr in Lake Vättern. 6. Contrary to earlier claims that crayfish predation on eggs of great Arctic charr is insignificant, our results indicate that crayfish predation may exceed fish predation and suggest that the abundance of signal crayfish on the spawning sites of great Arctic charr should be managed.