Deconstructing Ixodes ricinus: a partial matrix model allowing mapping of tick development,mortality and activity rates

A stage‐structured Leslie matrix model of a partial, discrete population of Ixodes ricinus (Linnaeus) (Ixodida: Ixodidae) ticks was developed to elucidate the impact of climate trends on the distribution and phenology of this species in the western Palaearctic. The model calculates development and mortality rates for each instar and evaluates recruitment rates based on the development of the tick population. The model captures the changes in development and mortality rates, providing a coherent index of performance correlated with the tick's geographic range. Maximum development rates are recorded for latitudes south of 36 °N and are spatially correlated with sites of maximum temperature, highest saturation deficit and highest mortality. The maximum available developmental time (the total annual time during which temperature allows development) for I. ricinus in the western Palaearctic is < 45% of the total year. North of 60 °N, available developmental time decreases sharply to only 15% of the year. The latitudinal boundary at which survival rates sharply drop is 43–46 °N, clearly delimiting the classically recognized extent of the main tick populations. The pattern of activity for larval–nymphal synchrony shows a clear west–east pattern. The model demonstrates the impact of climate according to tick stage and geographic location, and provides a practical framework for testing how the tick's lifecycle is affected by climate change.     

Targeting viral infection by microRNA inhibition

An inhibitor of microRNA-122 reduces viral load in chimpanzees that are chronically infected with hepatitis C virus, suggesting that such an approach might have therapeutic potential in humans.     

Cytogenetic assays of chemical clastogens using mammalian cells in culture.

A protocol is described for cytogenetic assays of chemical mutagens using mammalian cells in vitro. The system employs continuous drug treatment (3 concentrations) for up to 8 h and recovery-cell populations after pulse treatments with a high dose. Both direct fixation (for recording spindle anomalies in anaphase) and colcemid-hypotonic fixation (for reading metaphase chromosome aberrations) are used in order to estimate the effects of an agent as a mitotic poison and as a clastogen respectively. Some DNA intercalating dyes (acridine orange, quinacrine mustard, neutral red) were found to be highly clastogenic whereas others (quinacrine dihydrochloride, 33258 Hoechst) are not.     

Drought‐resistant fungi control soil organic matter decomposition and its response to temperature

Microbial‐mediated decomposition of soil organic matter (SOM) ultimately makes a considerable contribution to soil respiration, which is typically the main source of CO₂ arising from terrestrial ecosystems. Despite this central role in the decomposition of SOM, few studies have been conducted on how climate change may affect the soil microbial community and, furthermore, on how possible climate‐change induced alterations in the ecology of microbial communities may affect soil CO₂ emissions. Here we present the results of a seasonal study on soil microbial community structure, SOM decomposition and its temperature sensitivity in two representative Mediterranean ecosystems where precipitation/throughfall exclusion has taken place during the last 10 years. Bacterial and fungal diversity was estimated using the terminal restriction fragment length polymorphism technique. Our results show that fungal diversity was less sensitive to seasonal changes in moisture, temperature and plant activity than bacterial diversity. On the other hand, fungal communities showed the ability to dynamically adapt throughout the seasons. Fungi also coped better with the 10 years of precipitation/throughfall exclusion compared with bacteria. The high resistance of fungal diversity to changes with respect to bacteria may open the controversy as to whether future ‘drier conditions’ for Mediterranean regions might favor fungal dominated microbial communities. Finally, our results indicate that the fungal community exerted a strong influence over the temporal and spatial variability of SOM decomposition and its sensitivity to temperature. The results, therefore, highlight the important role of fungi in the decomposition of terrestrial SOM, especially under the harsh environmental conditions of Mediterranean ecosystems, for which models predict even drier conditions in the future.     

Novel nuclear methylation of sterols by the nematode Caenorhabditis elegans

Caenorhabditis elegans possesses a unique sterol methylation pathway not reported to occur in any other organism and also removes the C-24 ethyl group of sitosterol (a plant sterol). This nematode produced substantial quantities of 4 alpha-methyl-5 alpha-cholest-8(14)-en-3 beta-ol and smaller amounts of lophenol from dietary cholesterol, desmosterol or sitosterol. When C. elegans was propagated in media containing sitosterol plus 25-azacoprostane hydrochloride (25-aza-5 beta-cholestane hydrochloride), an inhibitor of delta 24-sterol reductase in insects, its 4 alpha-methylsterol fraction largely consisted of equal amounts of 4 alpha-methyl-5 alpha-cholesta-7,24-dien-3 beta-ol and 4 alpha-methyl-5 alpha-cholesta-8(14),24-dien-3 beta-ol. Thus 25-azacoprostane hydrochloride inhibited both a delta 24-sterol reductase and a delta 7-sterol isomerase in C. elegans.     

Comparison of known and suspected pheromonal constituents in males of the African ticks,Amblyomma hebraeum Koch andAmblyomma variegatum (Fabricius)

Three low molecular weight compounds were found in hexane: diethyl ether extracts of fed males of the African ticks,Amblyomma variegatum (tropical bont tick) andA. hebraeum (bont tick), namely,o-nitrophenol, methyl salicylate and 2,6-dichlorophenol. These same compounds were also found in a rinse of fedA. variegatum males, but were absent or present in only trace amounts in a rinse of fedA. hebraeum males.o-Nitrophenol and methyl salicylate were present in much higher concentrations (i.e., amounts/tick) inA. variegatum than inA. hebraeum. 2,6-Dichlorophenol was also more abundant inA. variegatum than inA. hebraeum, but the differences were not as great as with the former two compounds. Extraction in hexane over a 3-week period revealed four additional compounds, benzaldehyde, benzyl alcohol, benzothiazole and nonanoic acid. The first three compounds were found in males of both species; nonanoic acid was found only inA. hebraeum males. Published reports consistently show strong attraction byo-nitrophenol and methyl salicylate for both sexes of the two bont tick species; 2,6-dichlorophenol and benzaldehyde have been reported to be attractive to both sexes ofA. hebraeum. The possible roles of these compounds, as well as others occasionally reported fromA. hebraeum andA. variegatum, as components of the aggregation/attachment pheromone or other pheromones is discussed.Supported by Cooperative Agreement No. AFR-0435A-00-9084-00 with the U.S. Agency for International Development to the Department of Infectious Diseases, College of Veterinary Medicine, University of Florida, Gainesville, FL.     

Clenbuterol (planipart(trade mark)) for the postponement of parturition in cattle

Clenbuterol is a highly specific, long-acting (four to eight hours) beta-two sympathomimetic which causes bronchodilation and tocolysis (myometrial paralysis). The tocolytic effect was explored as a means to control parturition and reduce dystocia. Forty-six heifers were injected i.m. with either Clenbuterol or saline placebo in a randomly controlled experiment. Animals were treated when a cervical dilation of five centimeters or more was detected by vaginal examination. Length of first, second and third stages of parturition, ease of parturition, maternal pelvic area and calf viability were compared between treatment groups. Treatment with Clenbuterol increased (P<0.025; 119 vs 468 mins) the time heifers were in Stage I. However, the lengths of Stages II and III, pelvic area at birth and calf viability were not influenced by treatment. Diameter of the cervix at treatment was negatively related to the length of Stage I delay. Pelvic area also significantly affected the length of Stage II. Clenbuterol effectively delays Stage I of parturition with no adverse effects on the fetus or dam.     

A new approach to the Container Positioning Problem

In this paper the Container Positioning Problem is revisited. This problem arises at busy container terminals and requires one to minimize the use of block cranes in handling the containers that must wait at the terminal until their next means of transportation. We propose a new Mixed Integer Programming model that not only improves on earlier attempts at this problem, but also better reflects reality. In particular, the proposed model adopts a preference to reshuffle containers in line with a just-in-time concept, as it is assumed that data is more accurate the closer to a container’s scheduled departure the time is. Other important improvements include a reduction in the model size, and the ability of the model to consider containers initially at the terminal. In addition, we describe several classes of valid inequalities for this new formulation and present a rolling horizon based heuristic for solving larger instances of the problem. We show that this new formulation drastically outperforms previous attempts at the problem through a direct comparison on instances available in the literature. Furthermore, we also show that the rolling horizon based heuristic can further reduce the solution time on the larger of these instances as well as find acceptable solutions to much bigger, artificially generated, instances.     

Metabolism of [14C]cholesterol in Manduca sexta pupae: Isolation and identification of sterol sulfates,free ecdysteroids,and ecdysteroid acids

[¹⁴C]Cholesterol was injected into fifth-instar larvae of Manduca sexta, and the metabolites were isolated and identified from 8-day-old male and female pupae. A major portion of the metabolized cholesterol was esterified either with a sulfate group or with fatty acids. The predominant ecdysteroid metabolites were 20-hydroxyecdysone, 20,26-dihydroxyecdysone, 20-hydroxyecdysonoic acid, and 3-epi-20-hydroxyecdysonoic acid. Smaller amounts of ecdysteroids were identified as conjugates of 26-hydroxyecdysone, 3-epi-20-hydroxyecdysone, 20,26-dihydroxyecdysone, and its 3α-epimer. The metabolic profiles were similar for both male and female pupae. The two ecdysteroid acids were identified by nuclear magnetic resonance spectroscopy and chemical ionization mass spectrometry and by mass spectral analyses of their methyl esters. Detection of 3-epi-20-hydroxyecdysonoic acid as a major metabolite is significant, as its occurrence has been scarcely reported. 3-Epiecdysteroid acid formation is discussed as a possible ecdysteroid-inactivating pathway that may be operating specifically in lepidopterous insects or in particular developmental stages such as eggs or pupae.