Effects of organic acid treatments on small hive beetles, <Emphasis Type="Italic">Aethina tumida</Emphasis>, and the associated yeast <Emphasis Type="Italic">Kodamaea ohmeri</Emphasis>

In honeybee, Apis mellifera, colonies infested with larval and adult small hive beetles (SHB), hive material, and in particular honey, tends to ferment, probably due to SHB-associated yeasts such as the predominant Kodamaea ohmeri. Here, we test the effects of organic acids on K. ohmeri and on SHB-infested honey/pollen combs. Organic acids were applied at standard concentrations used by beekeepers to control other pests. In laboratory tests, the growth of K. ohmeri was significantly inhibited by lactic, formic and acetic acids. Treatments of SHB-infested honey/pollen combs (N = 18 colonies) with acetic acid significantly increased mortality of adult SHB and treatments with formic acid significantly reduced larval infestation. Our data suggest that treatment of honeybee colonies and storage rooms with organic acids could also help in reducing SHB damage.     

An empirical approach to target DNA quantification in environmental samples using real-time polymerase chain reactions

Recent advances in molecular techniques have allowed for the routine examination of nucleic acids in environmental samples. Although current methodologies are very sensitive, accurate target DNA quantification from environmental samples remains challenging. To facilitate high-throughput DNA quantification from environmental samples, we developed a novel DNA quantification method based on a non-linear curve-fitting approach to extract additional information from quantitative PCR amplification curves and used the fitted parameters to develop multiple regression standard equations for target DNA quantification. A 3-parameter sigmoidal function performed superior to a 4-parameter Weibull function for generating the multiple regression standard equations. In a verification experiment, target DNA was quantified in a series of ‘unknown’ samples in three soils using this approach and the results were compared to target DNA values determined using corrected and uncorrected Ct-based (threshold cycle) methods. For each method, the deviations from the expected target DNA content were determined. Results clearly showed that over all DNA concentrations, target DNA content determined by the non-linear curve-fitting method was more accurate and more precise than values predicted by all other methods. Analysis of variance conducted on the predicted DNA contents also revealed fewer statistical artifacts with the non-linear curve fitting method compared to the conventional Ct-based methods. The novel approach described here is accurate, inexpensive, and very amenable for automation and high-throughput applications.     

Surplus killing by introduced predators in Australia—evidence for ineffective anti-predator adaptations in native prey species?

Australian examples of surplus killing by mammalian predators were collated. These included surplus killing of native mammals and birds by foxes (Vulpes vulpes) and stock, native mammals and native birds by dingoes (Canis lupus dingo). We found no examples of surplus killing by feral cats (Felis catus). Incidents collated include historical anecdotes of surplus killing by foxes as they colonised the Australian mainland, recent examples where foxes killed threatened native species at sites despite intensive management to exclude foxes, and recent examples of the killing of native species on formerly fox-free islands to which foxes gained entry. Episodes of surplus killings by foxes, other than predation on captive or closely confined animals, appeared different in kind and frequency to those documented for co-evolved predator-prey systems on the large continental landmasses. They did not appear to be uncommon events associated with synchronised births of prey species, unusual or extreme weather that disadvantaged prey species, or seasonal food caching by a predator. Rather, surplus killing events appeared to reflect ineffective anti-predator defences by prey species when encountering a novel and efficient predator to which they have had no evolutionary exposure. We suggest that surplus killing by foxes may have been a feature of, and major contributor to, the rapid mainland extinction or contraction in range of many native species in Australia. In contrast to foxes, examples of surplus killing by dingoes relate mostly to domestic stock (calves and sheep). The arrival of dingoes to the Australian continent preceded that of foxes by 3500-4000 years, but they appear not to have had the dire impact on native mammals that we attribute to foxes. This may be due to fundamental differences in hunting styles and prey size and to their sparse populations in pre-European Australia. Active persecution of non-commensal dingoes by Aborigines, the lack of free-water, and the absence of European rabbits (Oryctolagus cuniculus) as an alternative food supply would have limited their numbers and their impact on native mammals.     

Sequence-directed DNA translocation by purified FtsK

DNA translocases are molecular motors that move rapidly along DNA using adenosine triphosphate as the source of energy. We directly observed the movement of purified FtsK, an Escherichia coli translocase, on single DNA molecules. The protein moves at 5 kilobases per second and against forces up to 60 piconewtons, and locally reverses direction without dissociation. On three natural substrates, independent of its initial binding position, FtsK efficiently translocates over long distances to the terminal region of the E. coli chromosome, as it does in vivo. Our results imply that FtsK is a bidirectional motor that changes direction in response to short, asymmetric directing DNA sequences.     

Spatial distribution of δ~2H and δ~18O values in the hydrologic cycle of the Nile Basin

Existing δ2H and δ18O values for precipitation and surface water in the Nile Basin were used to analyze precipitation inputs and the influence of evaporation on the isotopic signal of the Nile River and its tributaries. The goal of the data analysis was to better understand basin processes that influence seasonal streamflow for the source waters of the Nile River, because climate and hydrologic models have continued to produce high uncertainty in the prediction of precipitation and streamflow in the Nile Basin. An evaluation of differences in precipitation δ2H and δ18O values through linear regression and distribution analysis indicate variation by region and season in the isotopic signal of precipitation across the Nile Basin. The White Nile Basin receives precipitation with a more depleted isotopic signal compared to the Blue Nile Basin. The hot temperatures of the Sahelian spring produce a greater evaporation signal in the precipitation isotope distribution compared to precipitation in the Sahara/Mediterranean region, which can be influenced by storms moving in from the Mediterranean Sea. The larger evaporative effect is reversed for the two regions in summer because of the cooling of the Sahel from inflow of Indian Ocean monsoon moisture that predominantly influences the climate of the Blue Nile Basin. The regional precipitation isotopic signals convey to each region's streamflow, which is further modified by additional evaporation according to the local climate. Isotope ratios for White Nile streamflow are significantly altered by evaporation in the Sudd, but this isotopic signal is minimized for streamflow in the Nile River during the winter, spring and summer seasons because of the flow dominance of the Blue Nile. During fall, the contribution from the White Nile may exceed that of the Blue Nile, and the heavier isotopic signal of the White Nile becomes apparent. The variation in climatic conditions of the Nile River Basin provides a means of identifying mechanistic processes through changes in isotope ratios of hydrogen and oxygen, which have utility for separating precipitation origin and the effect of evaporation during seasonal periods. The existing isotope record for precipitation and streamflow in the Nile Basin can be used to evaluate predicted streamflow in the Nile River from a changing climate that is expected to induce further changes in precipitation patterns across the Nile Basin.     

Cardiopulmonary bypass in the cat

OBJECTIVE: To assess the physiologic response to, and acute survival of, cats undergoing cardiopulmonary bypass (CPB) and to evaluate the efficacy of a commercial human pediatric oxygenator system on cats weighing less than 6 kg. STUDY DESIGN: Experimental study. ANIMALS: Six intact male cats METHODS: Cats were placed on cardiopulmonary bypass by cannulating the cranial and caudal vena cavae and the carotid artery. The pediatric CPB circuit was primed with 150 mL of a balanced crystalloid solution. Venous drainage was enhanced by a controlled, vacuum-assist system. A cross-clamp was placed on the ascending aorta and cardiac arrest was induced by antegrade infusion of a cold cardioplegia solution. After 45 minutes of arrest time, the cross-clamp was removed and the cats were weaned off bypass and decannulated. No blood products were administered. Heart rate, mean arterial pressure (MAP), central venous pressure, arterial blood gas, hematocrit (HCT), total plasma protein concentration (TP), serum electrolyte concentrations, and activated clotting time (ACT) were measured at baseline period (BL), during CPB, 60 minutes after CPB (CPB 60) and 90 minutes after CPB (CPB 90). A complete blood count (CBC), blood chemistry profile, and urinalysis were performed at BL, during CPB, and CPB 90. Cats were euthanatized after CPB 90. RESULTS: Cardiopulmonary bypass resulted in a significant (P <.05) decrease in mean HCT (18.0%) and TP (2.3 gm/dL) at CPB 90 when compared to BL (30.5% and 6.0 gm/dL, respectively). The MAP at CPB 90 (54 mm Hg) was decreased from BL (94 mm Hg). The ACT increased from a mean of 124 seconds to > 400 seconds with heparinization and was reversed to 300 seconds with protamine. Mean platelet counts decreased from BL (369,000 /microL) to CPB 90 (94,500 /microL). Mean white blood cell counts decreased from 13,200 /microL at BL to 2,200 /microL at CPB 90. Upon reperfusion, 1 cat fibrillated but was successfully defibrillated. CONCLUSIONS: Cardiopulmonary bypass was performed successfully in 6 cats weighing less than 6 kg. Acute survival to 90 minutes after CPB was achieved in all 6 cats CLINICAL RELEVANCE: The ability to perform CPB in the cat may allow intracardiac repair of various heart defects in this species.     

Effect of glyphosate on the tripartite symbiosis formed by Glomus intraradices,Bradyrhizobium japonicum,and genetically modified soybean

Most soybeans grown in North America are genetically modified (GM) to tolerate applications of the broad-spectrum herbicide glyphosate; as a result, glyphosate is now extensively used in soybean cropping systems. Soybean roots form both arbuscular mycorrhizal (AM) and rhizobial symbioses. In addition to individually improving host plant fitness, these symbioses also interact to influence the functioning of each symbiosis, thereby establishing a tripartite symbiosis. The objectives of this study were to (1) estimate the effects of glyphosate on the establishment and functioning of AM and rhizobial symbioses with GM soybean, and (2) to estimate the interdependence of the symbioses in determining the response of each symbiosis to glyphosate. These objectives were addressed in two experiments; the first investigated the importance of the timing of glyphosate application in determining the responses of the symbionts and the second varied the rate of glyphosate application. Glyphosate applied at recommended field rates had no effect on Glomus intraradices or Bradyrhizobium japonicum colonization of soybean roots, or on soybean foliar tissue [P]. N₂-fixation was greater for glyphosate-treated soybean plants than for untreated-plants in both experiments, but only when glyphosate was applied at the first trifoliate soybean growth stage. These data deviate from previous studies estimating the effect of glyphosate on the rhizobial symbiosis, some of which observed negative effects on rhizobial colonization and/or N₂-fixation. We did observe evidence of the response of one symbiont (stimulation of N₂-fixation following glyphosate) being dependent on co-inoculation with the other; however, this interactive response appeared to be contextually dependent as it was not consistent between experiments. Future research needs to consider the role of environmental factors and other biota when evaluating rhizobial responses to herbicide applications.     

Ordered and dynamic assembly of single spliceosomes

The spliceosome is the complex macromolecular machine responsible for removing introns from precursors to messenger RNAs (pre-mRNAs). We combined yeast genetic engineering, chemical biology, and multiwavelength fluorescence microscopy to follow assembly of single spliceosomes in real time in whole-cell extracts. We find that individual spliceosomal subcomplexes associate with pre-mRNA sequentially via an ordered pathway to yield functional spliceosomes and that association of every subcomplex is reversible. Further, early subcomplex binding events do not fully commit a pre-mRNA to splicing; rather, commitment increases as assembly proceeds. These findings have important implications for the regulation of alternative splicing. This experimental strategy should prove widely useful for mechanistic analysis of other macromolecular machines in environments approaching the complexity of living cells.     

离子交换树脂膜原位提取土壤有效性养分的探讨

连续3年在实验室内和利用盆栽及田间进行了利用离子交换树脂膜对土壤的原位测定,探讨影响树脂膜对土壤中养分离子吸收的因素;制定出一套包括树脂膜埋置方法,时间,洗脱土粒和解吸被树脂膜吸附的养分等的标准方法,称为植物根模拟法(Plant root simulator)。 通过400多个土壤样品测定结果表明,用树脂膜提取的土壤有效氮、磷、钾和硫与用常规化学法提出的量有显著相关。盆栽条件下,树脂膜原位埋置法对预测供试作物油菜吸取土壤有效养分的能力与常规化学法相当,而硫的效果更好。树脂膜在田间直接埋置能合理地表征土壤中硝态氮的分布,并具有同时提取土壤各种有效养分的能力。由于离子树脂交换膜不受地域影响,具有广泛的通用性,因而在土壤养分有效性的研究中应用前景广阔。