A “Stripping” Ligand Tactic for Use with the Kinetic Locking-on Strategy: Its Use in the Resolution and Bioaffinity Chromatographic Purification of NAD-Dependent Dehydrogenases

The kinetic locking-on strategy utilizes soluble analogues of the target enzymes' specific substrate to promote selective adsorption of individual NAD⁺-dependent dehydrogenases on their complementary immobilized cofactor derivative. Application of this strategy to the purification of NAD⁺-dependent dehydrogenases from crude extracts has proven that it can yield bioaffinity systems capable of producing one-chromatographic-step purifications with yields approaching 100%. However, in some cases the purified enzyme preparation was found to be contaminated with other proteins weakly bound to the immobilized cofactor derivative through binary complex formation and/or nonspecific interactions, which continuously “dribbled” off the matrix during the chromatographic procedure. The fact that this problem can be overcome by including a short pulse of 5′-AMP (stripping ligand) in the irrigant a couple of column volumes prior to the discontinuation of the specific substrate analogue (locking-on ligand) is clear from the results presented in this report. The general effectiveness of this auxiliary tactic has been assessed using model studies and through incorporation into an actual purification from a crude cellular extract. The results confirm the usefulness of the stripping-ligand tactic for the resolution and purification of NAD⁺-dependent dehydrogenases when using the locking-on strategy. These studies have been carried out using bovine liver glutamate dehydrogenase (GDH, EC 1.4.1.3), yeast alcohol dehydrogenase (YADH, EC 1.1.1.1), porcine heart mitochondrial malate dehydrogenase (mMDH, EC 1.1.1.37), and bovine heart -lactate dehydrogenase ( -LDH, EC 1.1.1.27).     

Female mating status affects male mating tactic expression in the wolf spider Rabidosa punctulata

Males and females have conflicting interests on the frequency and outcomes of mating interactions. Males maximize their fitness by mating with as many females as possible, whereas choosy females often reduce receptivity following copulation. Alternative male mating tactics can be adaptive in their expression to a variety of mating contexts, including interactions with a relatively unreceptive mated female. Male Rabidosa punctulata wolf spiders can adopt distinctive mating tactics when interacting with a female, a complex courtship display, and/or a more coercive direct mount tactic that often involves grappling with females for copulation. In this study, we set up female mating treatments with initial trials and then paired mated and unmated females with males to observe both female remating frequencies and the male mating tactics used during the interactions. Males adopted different mating tactics depending on the mating status of the female they were paired with. Males were more likely to adopt a direct mount tactic with already-mated females and courtship with unmated females. Already-mated females were considerably less receptive to males during experimental trials, although they did remate 34% of the time, the majority of which were with males using a direct mount tactic. Whereas males adjusting to these contextual cues were able to gain more copulations, the observation of multiple mating in female R. punctulata introduces the potential for sperm competition. We discuss this sexual conflict in terms of the fitness consequences of these mating outcomes for both males and females.     

不同密度下和不同发情期时段中雄性麋鹿的繁殖计策

从1998年到2001年,我们研究了密度、发情期与雄性麋鹿繁殖计策的关系。我们选择了江苏大丰麋鹿保护区的两个麋鹿群体和北京麋鹿苑的一个麋鹿群体,观察记录了发情期前期、中期和后期中实验麋鹿种群参数、围栏大小、群体中雄鹿的繁殖计策、群主变更情况、后宫数目以及后宫雌鹿数。尽管不同种群和不同发情期的雄鹿繁殖计策有差异,但是各个群体中雄性麋鹿都有占群、偷袭交配和求偶集会3种繁殖计策。利用因子分析提取了两个主成分:主成分1主要代表实验围栏、麋鹿存栏数和密度等因子的影响,主成分2主要代表发情期时段因子的影响。这两个主成分解释了全部因素效应的90.72%。用Spearman相关分析发现,占群计策与主成分2呈极显著负相关,求偶集会计策与主成分1呈显著正相关,最大后宫雌鹿个体数与主成分1极显著正相关,而偷袭交配计策和后宫数目与主成分1和2均无显著相关。据此推测,占群和求偶集会繁殖计策分别受密度和发情期时段的影响,而偷袭交配行为对密度和发情期时段不敏感。我们还探讨了麋鹿发情行为计策的环境适应性,从行为学角度为麋鹿保护提供了新思路。     

Searching at different spatial scales: the foraging behaviour of the aphid parasitoid Aphidius rosae in rose bushes

The searching patterns of the aphid parasitoid Aphidius rosae were analysed at three different spatial scales: leaves, shoots and bushes. Parasitoid females searched aphid-infested leaves and shoots intensively and remained on average more than twice as long on infested than on uninfested shoots. Patch times and oviposition numbers per shoot were highly variable both between females and for different shoot visits within females. However, at the shoot and bush level low oviposition numbers were generally found. The time spent on different behavioural patterns (searching, resting, feeding, host handling) changed significantly during subsequent shoot visits of individual females but oviposition success was not influenced by this change. Parasitoids searched individual leaves and shoots mainly by walking, while moving between shoots occurred exclusively by flight. The travel time between shoots (i.e. flight time) accounted for less than 1% of the residence time in a bush. At the bush level foraging was characterized by a high ability to localize infested shoots and consequently little time was wasted in searching on uninfested shoots. The pattern of resource exploitation of individual females was consistent with the distribution of A. rosae larvae in field samples taken from individual rose bushes.     

Interspecific relationship between the Japanese horned beetle and two Japanese stag beetle species

The Japanese horned beetle Trypoxylus dichotomus septentrionalis and the Japanese stag beetles Lucanus maculifemoratus maculifemoratus and Prosopocoilus inclinatus inclinatus generally occur syntopically and aggregate on oak tree surfaces that exude sap. Securement of these sap sites might be directly linked to male reproductive success. Among the three species, it is likely that males of T. d. septentrionalis are dominant in occupation of feeding sites because of their larger body size. However, there is no clear evidence of this superiority. Moreover, if T. d. septentrionalis is dominant, the mechanism by which the two stag beetle species secure the feeding sites remains unclear. In the present study, I used body mass to compare the body size among males of T. d. septentrionalis, L. m. maculifemoratus and P. i. inclinatus. Further, to clarify the interspecific relationship between the horned beetle and the two stag beetles, I investigated the seasonality of emergence pattern of males at the feeding sites in the field. Comparison of body mass and observation of fighting behavior revealed that males of T. d. septentrionalis have an apparent superiority over males of the two stag beetle species. The seasonal emergence patterns of the two stag beetle species at the feeding sites showed bimodal distributions, and avoided the peak of emergence in T. d. septentrionalis. My results suggest that the two stag beetle species exhibit mate‐securing tactics by emerging at the feeding sites early and late during the breeding season, in order to avoid encountering T. d. septentrionalis.     

Disentangling Natural From Hunting Mortality in an Intensively Hunted Wild Boar Population

Abstract We assessed age-specific natural mortality (i.e., excluding hunting mortality) and hunting mortality of 1,175 male and 1,076 female wild boar (Sus scrofa) from Chǎteauvillain-Arc en Barrois (eastern France), using a 22-year dataset (1982–2004) and mark-recapture-recovery methods. Overall yearly mortality was >50% for all sex and age-classes. Low survival was mostly due to high hunting mortality; a wild boar had a >40% of chance of being harvested annually, and this risk was as high as 70% for adult males. Natural mortality rates of wild boar were similar for males and females (approx. 0.15). These rates were comparable to rates typical of male ungulates but high for female ungulates. Wild boar survival did not vary across sex and age-classes. Despite high hunting mortality, we did not detect evidence of compensatory mortality. Whereas natural mortality for males was constant over time, female mortality varied annually, independent of fluctuations in mast availability. Female wild boar survival patterns differed from those reported in other ungulates, with high and variable natural mortality. In other ungulates, natural mortality is typically low and stable across a wide range of environmental conditions. These differences may partly reflect high litter sizes for wild boar, which carries high energetic costs. High hunting mortality may induce a high investment of females in reproduction early in life, at the detriment to survival. Despite high hunting mortality, the study population increased. Effective population control of wild boar should target a high harvest rate of piglets and reproductive females.     

微藻趋向运动及其靶向运输应用研究进展

微藻主要是指一类能进行光合自养的微生物,且很多藻株还兼具运动特性.因而将微藻与微流控芯片结合,实现精确靶向,或用于分子药物递送,在生物医学治疗和药效学分析等领域有重要的潜在应用价值,也是当今的研究热点之一.然而,目前关于微藻趋向运动的研究及潜在的应用的综述报道却相对较少.本文主要以模式微藻莱茵衣藻为例,概述基于微藻细胞...     

Configuration of daily grazing and searching of growing beef cattle in grassland: observational study

Many of the studies in Campos grasslands focus on management aspects such as the control of herbage allowance, and application of nutrients and/or overseeding with legumes. However, there is little literature on how the Campos grassland resource is utilised, especially regarding the grazing pattern and the relationship between pasture quantity and quality on daily grazing activities. The study of the ingestive behaviour in species-rich and heterogeneous native grasslands during daylight hours, and understanding how animals prioritise quality or quantity of intake in relation to pasture attributes, are important to comprehend the ingestive-digestive processes modulating the energy intake of animals and to achieve a better grazing management. Therefore, the objective was to describe and quantify the daily grazing behaviour of growing cattle grazing native pasture with different structures as a result of different management practices, and study the relationship of pasture attributes and intake through multivariate analysis. The study was carried out at the Faculty of Agronomy, Paysandú, Uruguay. Treatments were native grassland, overseeding with Trifolium pratense and Lotus tenuis + phosphorus, and native pasture + nitrogen-phosphorus. Grazing activities were discriminated into grazing, searching (defined when animals take 1–2 bites in one feeding station and then change to another feeding station and so on), ruminating and idling. The probability of time allocated to each activity was continuously measured during daylight hours (0700–1930) and was related to pasture structure and forage quality using regression tree models, while the bite rate was determined every 2 h. The diurnal pattern of growing cattle showed grazing and searching sessions, followed by ruminating and idling sessions. The length of sessions (as the probability of time allocated to each activity) varied throughout the day. The grazing probability was greater during afternoon than morning and midday (0.74 vs 0.45 vs 0.46, respectively), and it was associated with higher bite rate (34.2 bites/min). Regression tree models showed different grazing, searching and ruminating strategies according to pasture attributes. During the morning, animals modified grazing, searching, ruminating and idling strategies according to bite rate, crude protein in diet and herbage allowance. At midday, they only adjusted ruminating and idling, while during afternoon sessions, grazing activities were modified by pasture quantity attributes such as herbage mass and herbage allowance. By controlling the herbage allowance, herbage mass and pasture height, animals prioritise quality in the morning and quantity in the afternoon, integrating and modifying the grazing-searching and ruminating-idling pattern.