Functional responses of the intertidal amphipod Echinogammarus marinus: effects of prey supply, model selection and habitat complexity.

The influence of predation in structuring ecological communities can be informed by examining the shape and magnitude of the functional response of predators towards prey. We derived functional responses of the ubiquitous intertidal amphipod Echinogammarus marinus towards one of its preferred prey species, the isopod Jaera nordmanni. First, we examined the form of the functional response where prey were replaced following consumption, as compared to the usual experimental design where prey density in each replicate is allowed to deplete. E. marinus exhibited Type II functional responses, i.e. inversely density-dependent predation of J. nordmanni that increased linearly with prey availability at low densities, but decreased with further prey supply. In both prey replacement and non-replacement experiments, handling times and maximum feeding rates were similar. The non-replacement design underestimated attack rates compared to when prey were replaced. We then compared the use of Holling’s disc equation (assuming constant prey density) with the more appropriate Rogers’ random predator equation (accounting for prey depletion) using the prey non-replacement data. Rogers’ equation returned significantly greater attack rates but lower maximum feeding rates, indicating that model choice has significant implications for parameter estimates. We then manipulated habitat complexity and found significantly reduced predation by the amphipod in complex as opposed to simple habitat structure. Further, the functional response changed from a Type II in simple habitats to a sigmoidal, density-dependent Type III response in complex habitats, which may impart stability on the predator−prey interaction. Enhanced habitat complexity returned significantly lower attack rates, higher handling times and lower maximum feeding rates. These findings illustrate the sensitivity of the functional response to variations in prey supply, model selection and habitat complexity and, further, that E. marinus could potentially determine the local exclusion and persistence of prey through habitat-mediated changes in its predatory functional responses.

Construction and evaluation of plasmid vectors optimized for constitutive and regulated gene expression in Burkholderia cepacia complex isolates

Genetic studies with Burkholderia cepacia complex isolates are hampered by the limited availability of cloning vectors and by the inherent resistance of these isolates to the most common antibiotics used for genetic selection. Also, some of the promoters widely employed for gene expression in Escherichia coli are inefficient in B. cepacia. In this study, we have utilized the backbone of the vector pME6000, a derivative of the pBBR1 plasmid that was originally isolated from Bordetella bronchiseptica, to construct a set of vectors useful for gene expression in B. cepacia. These vectors contain either the constitutive promoter of the S7 ribosomal protein gene from Burkholderia sp. strain LB400 or the arabinose-inducible P(BAD) promoter from E. coli. Promoter sequences were placed immediately upstream of multiple cloning sites in combination with the minimal sequence of pME6000 required for plasmid maintenance and mobilization. The functionality of both vectors was assessed by cloning the enhanced green fluorescent protein gene (e-gfp) and determining the levels of enhanced green fluorescent protein expression and fluorescence emission for a variety of clinical and environmental isolates of the B. cepacia complex. We also demonstrate that B. cepacia carrying these constructs can readily be detected intracellularly by fluorescence microscopy following the infection of Acanthamoeba polyphaga.

Construction and biochemical characterization of recombinant cytoplasmic forms of the IucD protein (lysine:N6-hydroxylase) encoded by the pColV-K30 aerobactin gene cluster

The aerobactin gene cluster in pColV-K30 consists of five genes (iucABCD iutA); four of these (iucABCD) are involved in aerobactin biosynthesis, whereas the fifth one (iutA) encodes the ferriaerobactin outer membrane receptor. iucD encodes lysine:N6-hydroxylase, which catalyzes the first step in aerobactin biosynthesis. Regardless of the method used for cell rupture, we have consistently found that IucD remains membrane bound, and repeated efforts to achieve a purified and active soluble form of the enzyme have been unsuccessful. To circumvent this problem, we have constructed recombinant IucD proteins with modified amino termini by creating three in-frame gene fusions of IucD to the amino-terminal amino acids of the cytoplasmic enzyme beta-galactosidase. Two of these constructs resulted in the addition to the iucD coding region of a hydrophilic leader sequence of 13 and 30 amino acids. The other construct involved the deletion of the first 47 amino acids of the IucD amino terminus and the addition of 19 amino acids of the amino terminus of beta-galactosidase. Cells expressing any of the three recombinant IucD forms were found to produce soluble N6-hydroxylysine. One of these proteins, IucD439, was purified to homogeneity from the soluble fraction of the cell lysates, and it was capable of participating in the biosynthesis of aerobactin, as determined in vitro by a cell-free system and in vivo by a cross-feeding bioassay. A medium ionic strength of 0.25 (250 mM NaCl) or higher was required to maintain the protein in a catalytically functional, tetrameric state.(ABSTRACT TRUNCATED AT 250 WORDS)

Confined Site Construction and its Impact on Personnel Productivity

Inner city, confined site construction is quickly becoming the norm within the construction sector. The aim of this paper is to identify and document the effect, if any, that a confined construction site environment has on the productivity of on-site personnel. In order to compile the relevant information and attain appropriate results on the matter in question, a qualitative analytical approach is adopted. This process incorporates multiple cases studies from Ireland, Northern Ireland and USA. From the resulting case studies, a minimum of three individual interviews and focus group seminars are conducted to aid in the collection of the data while also assisting in the confirmation of the factors identified from a critique of the relevant literature. From the resulting case studies and discussions, a list of the key issues pertaining to the on-site productivity of personnel emerged and is documented as follows; 1) Overcrowding of personnel at workstations, 2) Lack of space for the effective movement of personnel on-site, 3) Numerous trades working within the one space on-site. Through identifying the issues highlighted and proactively mitigating or eliminating the factors detailed, on-site management professionals can strive to ensure maximum productivity from the industry’s most important resource – people.

Environmental Aspects of Underground Construction and Exploration of Underground Structure – State of the Art

An underground work (such as a tunnel or a cavern) has many, well known, environmental qualities such as: no physical barriers crossing the land, less maintenance costs than an analogous surface structure, less expenses for heating and conditioning; a localized emission of noise, gas, dust during operation and, finally, a better protection against seismic actions.
It cannot be forgotten, anyway, that some negative environmental features are present such as, for example, : perturbation, pollution and drainage of the groundwater; settlements; disposal of waste rock.
In the paper the above mentioned concepts are discussed and analysed to give a global overview of all this aspects.

Lovastatin biosynthesis depends on the carbon-nitrogen proportion: model development and controller design

Lovastatin biosynthesis depends on the relative concentrations of dissolved oxygen and the carbon and nitrogen resources. An elucidation of the underlying relationship would facilitate the derivation of a controller for the improvement of lovastatin yield in bioprocesses. To achieve this goal, batch submerged cultivation experiments of lovastatin production by Aspergillus flavipus BICC 5174, using both lactose and glucose as carbon sources, were performed in a 7 liter bioreactor and the data used to determine how the relative concentrations of lactose, glucose, glutamine and oxygen affected lovastatin yield. A model was developed based on these results and its prediction was validated using an independent set of batch data obtained from a 15-liter bioreactor using five statistical measures, including the Willmott index of agreement. A nonlinear controller was designed considering that dissolved oxygen and lactose concentrations could be measured online, and using the lactose feed rate and airflow rate as process inputs. Simulation experiments were performed to demonstrate that a practical implementation of the nonlinear controller would result in satisfactory outcomes. This is the first model that correlates lovastatin biosynthesis to carbon-nitrogen proportion and possesses a structure suitable for implementing a strategy for controlling lovastatin production.

Multibody modelling of a TB31 and a TB32 crash test with vertical portable concrete barriers: model verification and sensitivity analysis.

In this article the multibody simulation software package MADYMO for analysing and optimizing occupant safety design was used to model crash tests for Normal Containment barriers in accordance with EN 1317. The verification process was carried out by simulating a TB31 and a TB32 crash test performed on vertical portable concrete barriers and by comparing the numerical results to those obtained experimentally. The same modelling approach was applied to both tests to evaluate the predictive capacity of the modelling at two different impact speeds. A sensitivity analysis of the vehicle stiffness was also carried out. The capacity to predict all of the principal EN1317 criteria was assessed for the first time: the acceleration severity index, the theoretical head impact velocity, the barrier working width and the vehicle exit box. Results showed a maximum error of 6% for the acceleration severity index and 21% for theoretical head impact velocity for the numerical simulation in comparison to the recorded data. The exit box position was predicted with a maximum error of 4°. For the working width, a large percentage difference was observed for test TB31 due to the small absolute value of the barrier deflection but the results were well within the limit value from the standard for both tests. The sensitivity analysis showed the robustness of the modelling with respect to contact stiffness increase of ±20% and ±40%. This is the first multibody model of portable concrete barriers that can reproduce not only the acceleration severity index but all the test criteria of EN 1317 and is therefore a valuable tool for new product development and for injury biomechanics research.

Arsenic Speciation in Food and Estimation of the Dietary Intake of Inorganic Arsenic in a Rural Village of West Bengal, India

Arsenic (As) species were quantified by HPLC-HG-AFS in water and vegetables from a rural area of West Bengal (India). Inorganic species predominated in vegetables (including rice) and drinking water; in fact, inorganic arsenic (i-As) represented more than 80% of the total arsenic (t-As) content. To evaluate i-As intake in an arsenic affected rural village, a food survey was carried out on 129 people (69 men and 60 women). The data from the survey showed that the basic diet, of this rural population, was mainly rice and vegetables, representing more than 50% of their total daily food intake. During the periods when nonvegetarian foods (fish and meat) were scarce, the importance of rice increased, and rice alone represented more than 70% of the total daily food intake. The food analysis and the food questionnaires administrated led us to establish a daily intake of i-As of about 170 mu g i-As day(-1), which was above the tolerable daily intake of 150 mu g i-As day(-1), generally admitted. Our results clearly demonstrated that food is a very important source of i-As and that this source should never be forgotten in populations depending heavily on vegetables (mainly rice) for their diet.

Note. Speciation of arsenic in licorice confectionery products and estimation of health risks

Total-arsenic (T-As) and arsenic (As) species were determined by HPLC-HG-AAS in ten different confectionery products: nine throat pearls and an industrial licorice extract. The Spanish legislation sets a maximum total-As content in confectionery products at 0.1 mu g/g. T-As concentrations were above the permitted maximum limit (mean of 0.219 +/- 0.008 mu g/g). All As was present in the form of toxic inorganic species. The daily consumption of licorice-confections in Spain is 1.1 g and leads to a daily intake of inorganic-As of 0.23 mu g (0.2% of the tolerable daily intake of inorganic As for a teenager). These experimental results proved that even though high total-As concentrations were found in licorice throat pearls and that all the As found was present as inorganic species, no significant risks for health are expected just by considering this As source.