Surface Sampling of Spores in Dry-Deposition Aerosols

The ability to reliably and reproducibly sample surfaces contaminated with a biological agent is a critical step in measuring the extent of contamination and determining if decontamination steps have been successful. The recovery operations following the 2001 attacks with Bacillus anthracis spores were complicated by the fact that no standard sample collection format or decontamination procedures were established. Recovery efficiencies traditionally have been calculated based upon biological agents which were applied to test surfaces in a liquid format and then allowed to dry prior to sampling tests, which may not be best suited for a real-world event with aerosolized biological agents. In order to ascertain if differences existed between air-dried liquid deposition and biological spores which were allowed to settle on a surface in a dried format, a study was undertaken to determine if differences existed in surface sampling recovery efficiencies for four representative surfaces. Studies were then undertaken to compare sampling efficiencies between liquid spore deposition and aerosolized spores which were allowed to gradually settle under gravity on four different test coupon types. Tests with both types of deposition compared efficiencies of four unique swabbing materials applied to four surfaces with various surface properties. Our studies demonstrate that recovery of liquid-deposited spores differs significantly from recovery of dry aerosol-deposited spores in most instances. Whether the recovery of liquid-deposited spores is overexaggerated or underrepresented with respect to that of aerosol-deposited spores depends upon the surface material being tested.     

Evaluation of Two Surface Sampling Methods for Detection of Erwinia herbicola on a Variety of Materials by Culture and Quantitative PCR

This research was designed to evaluate surface sampling protocols for use with culture and quantitative PCR (QPCR) amplification assay for detection of the gram-negative bacterial biothreat simulant Erwinia herbicola on a variety of surface materials. Surfaces selected for evaluation were wood laminate, glass and computer monitor screens, metal file cabinets, plastic arena seats, nylon seat cushions, finished concrete flooring, and vinyl tile flooring. Laboratory and test chamber studies were performed to evaluate two sampling methods, a sponge and a macrofoam swab, for detection of E. herbicola on surface materials. In laboratory trials, seven materials were inoculated with a known concentration of E. herbicola cells and samples were collected from the surfaces of the materials to determine sampling efficiencies. Culture analysis was ineffective for assessing E. herbicola collection efficiency because very few culturable cells were obtained from surface samples. QPCR demonstrated that E. herbicola DNA was present in high concentrations on all of the surface samples, and sampling efficiencies ranged from 0.7 to 52.2%, depending on the sampling method and the surface material. The swab was generally more efficient than the sponge for collection of E. herbicola from surfaces. Test chamber trials were also performed in which E. herbicola was aerosolized into the chamber and allowed to settle onto test materials. Surface sampling results supported those obtained in laboratory trials. The results of this study demonstrate the capabilities of QPCR to enhance the detection and enumeration of biocontaminants on surface materials and provide information on the comparability of sampling methods.     

Evaluation of two surface sampling methods for detection of Erwinia herbicola on a variety of materials by culture and quantitative PCR

This research was designed to evaluate surface sampling protocols for use with culture and quantitative PCR (QPCR) amplification assay for detection of the gram-negative bacterial biothreat simulant Erwinia herbicola on a variety of surface materials. Surfaces selected for evaluation were wood laminate, glass and computer monitor screens, metal file cabinets, plastic arena seats, nylon seat cushions, finished concrete flooring, and vinyl tile flooring. Laboratory and test chamber studies were performed to evaluate two sampling methods, a sponge and a macrofoam swab, for detection of E. herbicola on surface materials. In laboratory trials, seven materials were inoculated with a known concentration of E. herbicola cells and samples were collected from the surfaces of the materials to determine sampling efficiencies. Culture analysis was ineffective for assessing E. herbicola collection efficiency because very few culturable cells were obtained from surface samples. QPCR demonstrated that E. herbicola DNA was present in high concentrations on all of the surface samples, and sampling efficiencies ranged from 0.7 to 52.2%, depending on the sampling method and the surface material. The swab was generally more efficient than the sponge for collection of E. herbicola from surfaces. Test chamber trials were also performed in which E. herbicola was aerosolized into the chamber and allowed to settle onto test materials. Surface sampling results supported those obtained in laboratory trials. The results of this study demonstrate the capabilities of QPCR to enhance the detection and enumeration of biocontaminants on surface materials and provide information on the comparability of sampling methods.     

Validation of a Nylon-Flocked-Swab Protocol for Efficient Recovery of Bacterial Spores from Smooth and Rough Surfaces

In order to meet planetary-protection requirements, culturable bacterial spore loads are measured representatively for the total microbial contamination of spacecraft. However, the National Aeronautics and Space Administration''s (NASA''s) cotton swab protocols for spore load determination have not changed for decades. To determine whether a more efficient alternative was available, a novel swab was evaluated for recovery of different Bacillus atrophaeus spore concentrations on stainless steel and other surfaces. Two protocols for the nylon-flocked swab (NFS) were validated and compared to the present NASA standard protocol. The results indicate that the novel swab protocols recover 3- to 4-fold more (45.4% and 49.0% recovery efficiency) B. atrophaeus spores than the NASA standard method (13.2%). Moreover, the nylon-flocked-swab protocols were superior in recovery efficiency for spores of seven different Bacillus species, including Bacillus anthracis Sterne (recovery efficiency, 20%). The recovery efficiencies for B. atrophaeus spores from different surfaces showed a variation from 5.9 to 62.0%, depending on the roughness of the surface analyzed. Direct inoculation of the swab resulted in a recovery rate of about 80%, consistent with the results of scanning electron micrographs that allowed detailed comparisons of the two swab types. The results of this investigation will significantly contribute to the cleanliness control of future life detection missions and will provide significant improvement in detection of B. anthracis contamination for law enforcement and security efforts.The recent discovery of liquid water on Mars has sparked debate about the possibility of extraterrestrial life (37). Consequently, highly sensitive biosensors will be deployed onboard spacecraft like the Mars Science Laboratory (MSL), using technologies such as gas chromatographical analysis to search for the smallest traces of life (http://mars.jpl.nasa.gov/msl/mission/). Contamination of equipment by terrestrial microorganisms resulting from a lack of spacecraft cleanliness could significantly compromise the integrity of life detection missions and result in falsely positive extraterrestrial life signals. The prevention of this so-called “forward contamination” is one major goal of American and European space agencies'' planetary-protection efforts. Regular determination of a spacecraft''s bioload and the mission components throughout assembly are mandatory for detecting unacceptably high contamination that exceeds levels set by the United Nations treaty (Outer Space Treaty [11]).Modern spacecraft hardware is very susceptible to standard heat sterilization protocols, so baking the entire spacecraft, such as the Viking Lander Capsule at 111.7°C ± 1.7°C for 23 to 30 h is no longer feasible (30). Alternative cleaning and sterilization methodologies for spacecraft components prior to assembly (i.e., nonthermal plasma technologies) have been discussed (36). However, after integration, sterile hardware is exposed to a significant risk of contamination during assembly, testing, and launching operations. Because of limited access to integrated spacecraft components, the microbial cleanliness of a spacecraft and its surroundings is meticulously maintained through frequent cleaning and sterilization routines. Therefore, the regular and frequent detection of possible contaminants in the assembly environment is more important than ever.To estimate the severity of microbial contamination, the National Aeronautics and Space Administration''s (NASA''s) standard procedure focuses on aerobic, mesophilic spores (26). Briefly, surface samples are taken from spacecraft using moist cotton swabs or wipes. After an extraction procedure, the samples are subjected to a short heat shock (15 min; 80°C) to kill vegetative cells and then pour plated in Trypticase soy agar (TSA) for the enumeration of CFU. This protocol was originally developed for the Viking mission more than 3 decades ago (30) and has remained, for the most part, unchanged.Recent studies have shown that cotton swabs have acceptable recovery efficiencies for Bacillus spores (41.7%) (32) but, due to their organic nature, may raise residue problems on surfaces. Furthermore, their comparatively high DNA content could lead to false positives or inhibition should NASA one day incorporate molecular technologies into their microbial-detection protocols (7).Based on these observations, researchers are beginning to move away from cotton in favor of alternative swabs made from rayon or macrofoam (6, 18). A recent study reported high recovery efficiencies for various vegetative cells from stainless steel surfaces by applying a novel swab with a bulb-shaped head flocked with nylon fibers (12). Patented in 2004, this design facilitates the release of particulates and microbes, resulting in a significantly higher detection rate. The broad applicability of these nylon-flocked swabs (NFS) has been demonstrated by their use in various clinical studies isolating pathogens from medical environments (1, 10, 20).General studies on surface-sampling tools have clearly shown that the swab material and the extraction method are the dominant factors in spore recovery efficiencies (32). Additionally, the properties of the surface to be sampled affect sample recovery (8). For planetary-protection applications, the broad variety of novel materials used in spacecraft construction must be considered. The Mars Exploration Rover mission craft, for example, was composed of at least five kinds of surface materials (http://marsrovers.jpl.nasa.gov/overview). While the cruise stage was constructed primarily of aluminum and the aeroshell consisted of aluminum honeycomb structures, the lander itself was made of titanium and graphite composite (carbon fiber-reinforced plastic [CFRP]). The airbag and the parachutes were made of Vectran and polyester/nylon fabrics. These different materials are quite challenging for sampling tools. Accurate sampling of materials with various surface textures will require planetary-protection programs to introduce novel swab materials.To our knowledge, no investigations have been performed to compare the recovery of spores from different spacecraft surfaces. Previous studies have compared cotton and synthetic sampling materials, but only on stainless steel surfaces (19), and no studies have compared sampling methods on actual spacecraft materials (7).Recently published protocols for spore detection have been based on one specific Bacillus species and/or on one type of surface. Unfortunately, these protocols provide no insight into the effects of varying these factors (4-6, 8, 9, 14, 18), as requested by USP (United States Pharmacopeia) 1223 for validation of alternative microbial methods (3). Some of the aforementioned studies were conducted in response to B. anthracis terrorism incidents in 2001 and used B. atrophaeus as a surrogate. Consequently, information about the actual sampling efficiency of B. anthracis spores is quite limited and may vary significantly from the B. atrophaeus data.In this comprehensive study, we evaluated the novel nylon-flocked swab and a corresponding protocol to recover Bacillus spores from five different spacecraft-related surfaces. It should be noted that although stainless steel served as the standard test surface, it is not a predominant material in spacecraft; however, since the majority of previous (sampling) studies were performed on stainless steel, it represents a universally recognized carrier and also serves as a conservative proxy for the average roughness of the materials used in space science.Our nylon-flocked-swab protocol was validated with respect to accuracy, precision, limit of detection, linearity, and robustness (3). Moreover, its specificity was determined by applying spores of seven different Bacillus species, including the avirulent, attenuated strain Bacillus anthracis Sterne, and by comparing the resulting recovery efficiencies. The results in this communication will significantly contribute to planetary-protection protocols and could also be of high interest for public health issues.     

Theoretical effects of feed composition, feed conversion and feed spillage on waste discharge in fish culture

Based on feed composition, fish composition and feed conversion, some theoretical effects of feeding on waste discharge are calculated to illustrate the appropriateness of farm effluent monitoring. The effects of feed conversion and feed spillage and feed composition on waste discharge (suspended and dissolved N, P & COD) are demonstrated. Slight variations in feed conversion at the moment of sampling, especially those evoked by feed spillage, have tremendous consequences for discharge values. It appears crucial to determine whether observed conversions are explained by inherent efficiencies of fish growth, or whether higher growth efficiencies are combined with feed spillage. Therefore, thorough knowledge of relationships between nutrient intake and growth should be applied to effluent assessment. A calculation is presented to serve as a preliminary assessment of the impacts of these variables on water quality monitoring. The concept is illustrated using measurements from a recirculation pilot eel culture system.     

Molecular biomass and MetaTaxogenomic assessment of soil microbial communities as influenced by soil DNA extraction procedure

Three soil DNA extraction procedures (homemade protocols and commercial kit) varying in their practicability were applied to contrasting soils to evaluate their efficiency in recovering: (i) soil DNA and (ii) bacterial diversity estimated by 16S rDNA pyrosequencing. Significant differences in DNA yield were systematically observed between tested procedures. For certain soils, 10 times more DNA was recovered with one protocol than with the others. About 15 000 sequences of 16S rDNA were obtained for each sample which were clustered to draw rarefaction curves. These curves, as well as the PCA ordination of community composition based on OTU clustering, did not reveal any significant difference between procedures. Nevertheless, significant differences between procedures were highlighted by the taxonomic identification of sequences obtained at the phylum to genus levels. Depending on the soil, differences in the number of genera detected ranged from 1% to 26% between the most and least efficient procedures, mainly due to a poorer capacity to recover populations belonging to Actinobacteria, Firmicutes or Crenarchaeota. This study enabled us to rank the relative efficiencies of protocols for their recovery of soil molecular microbial biomass and bacterial diversity and to help choosing an appropriate soil DNA extraction procedure adapted to novel sequencing technologies.     

Towards robust and repeatable sampling methods in eDNA‐based studies

DNA‐based techniques are increasingly used for measuring the biodiversity (species presence, identity, abundance and community composition) of terrestrial and aquatic ecosystems. While there are numerous reviews of molecular methods and bioinformatic steps, there has been little consideration of the methods used to collect samples upon which these later steps are based. This represents a critical knowledge gap, as methodologically sound field sampling is the foundation for subsequent analyses. We reviewed field sampling methods used for metabarcoding studies of both terrestrial and freshwater ecosystem biodiversity over a nearly three‐year period (n = 75). We found that 95% (n = 71) of these studies used subjective sampling methods and inappropriate field methods and/or failed to provide critical methodological information. It would be possible for researchers to replicate only 5% of the metabarcoding studies in our sample, a poorer level of reproducibility than for ecological studies in general. Our findings suggest greater attention to field sampling methods, and reporting is necessary in eDNA‐based studies of biodiversity to ensure robust outcomes and future reproducibility. Methods must be fully and accurately reported, and protocols developed that minimize subjectivity. Standardization of sampling protocols would be one way to help to improve reproducibility and have additional benefits in allowing compilation and comparison of data from across studies.     

Influence of sampling protocol on diet determination of gentoo penguins Pygoscelis papua and Antarctic fur seals Arctocephalus gazella

The influence of two sampling protocols on diet determination of two marine predators, the gentoo penguin (Pygoscelis papua) and Antarctic fur seal (Arctocephalus gazella), was investigated. The collection of diet samples on three occasions over a 2-week period was compared with collecting all samples during a single session, as current CCAMLR Ecosystem Monitoring Programme protocols recommend. Some differences in the mass of food recovered from penguins were found but this was attributed to the mass of penguin sampled. There were no differences in diet composition between protocols and although body mass was a significant determinant of the mean length of krill Euphausia superba recovered from penguins, there were no differences between sampling protocols. This study has shown that differences between sampling frequencies are small and a variety of sampling protocols can produce results acceptable for inter-annual monitoring. The mass of sampled individuals can account for significant variation and should be recorded, especially if sampling frequencies and sizes are low. Accepted: 24 March 1999     

A Critical Comparison of Two Long-term Zooplankton Time Series from the Central-west North Sea

Long-term changes in relative and absolute zooplankton abundanceand species composition were compared between the Dove MarineLaboratory and Continuous Plankton Recorder (CPR) time seriesin the central-western North Sea. The two time series employdifferent sampling mechanisms, with the Dove series beng obtainedby net sampling at a fixed station off the Northumberland coast,while the CPR series utilizes a network of fixed towed routes. It was found that there was a significant correlation (r =0.64, P < 0.001) between the relative year-to-year fluctuations of the two series and a significant agreement in the pattern of year-to-year variations in species composition of the dominant taxa (P = 0.01) over the majority of the time period. However, examination of absolute zooplankton abundances found large differences between the two time series. Differences in mesh size and in the sample processing methodologies were not wholly responsible for these. Model-derived catching efficiencies for the two sampling devices suggested that differences in absolute abundances may have been mainly due in some taxa to a greater degree of active avoidance of the CPR sampling device by zooplankton, although it is likely that passive avoidance as a result of hydrodynamic factors has a role.>     

Validation of the human T-lymphocyte cloning assay — ring test report from the EU concerted action on HPRT mutation (EUCAHM)

The T-cell cloning assay, which enables the enumeration and molecular analysis of 6-thioguanine resistant (HPRT-negative) mutant T-cells, has been extensively used for studying human somatic gene mutation in vivo. However, large inter-laboratory variations in the HPRT mutant frequency (MF) call for further investigation of inter-laboratory differences in the experimental methodology, and development of an optimal but easy uniform cloning protocol. As part of the EU Concerted Action on HPRT Mutation (EUCAHM), we have carried out two Ring tests for the T-cell cloning assay. For each test, duplicate and coded samples from three buffy coats were distributed to five laboratories for determination of MF using six different protocols. The results indicated a good agreement between split samples within each laboratory. However, both the cloning efficiencies (CEs) and MFs measured for the same blood donors showed substantial inter-laboratory variations. Also, different medium compositions used in one and the same laboratory resulted in a remarkable difference in the level of MF. A uniform operating protocol (UOP) was proposed and compared with the traditional protocols in the second Ring test. The UOP (preincubation) increased the CE in laboratories traditionally using preincubation, but decreased the CE in laboratories traditionally using priming. Adjusted for donor, use of different protocols contributed significantly to the overall variation in lnCE (P=0.0004) and lnMF (P=0.03), but there was no significant laboratory effect on the lnCE (P=0.38) or lnMF (P=0.14) produced by the UOP alone. Finally, a simplified version of the UOP using the serum-free medium X-Vivo 10 and PMA was tested in one laboratory, and found to produce a considerable increase in CE. This modified UOP needs to be further evaluated in order to be used for future databases on HPRT MFs in various populations.     

Simple,rapid and reliable methods to obtain high quality RNA and genomic DNA from <Emphasis Type="Italic">Quercus ilex</Emphasis> L. leaves suitable for molecular biology studies

Isolation of high-quality RNA and genomic DNA (gDNA) from many samples is a necessary step before accomplishing molecular biology studies. The particular composition of Quercus ilex leaves, specially hard and rich in cell wall material, polyphenolics and secondary metabolites, usually results in preparations contaminated with non-nucleic acid compounds. Although many methods have been developed, each case of study demands a protocol adapted to the specific plant sample and the pursued research objectives. We have evaluated several protocols to establish the methodology that best suited to our current genetic and molecular studies on Q. ilex. Our priority was to select the simplest methods reducing the plant starting material and the time employed, without compromising yield, quality and integrity of the isolated nucleic acids. Our results point to two protocols based on silica-membrane purification, as the most convenient for Q. ilex leaf tissue, and both procedures are greatly improved by adding insoluble polyvinyl polypyrrolidone during the isolation process. The protocols optimized here can be completed at the microfuge scale and allow a researcher to process 48 samples in 1 h, producing high quality preparations suitable for the routinely molecular biology applications with higher efficiency than other more labour and time-consuming protocols.     

Response surface methods for optimizingSaccharopolyspora spinosa,a novel macrolide producer

Summary Strain A83543, recently identified asSaccharopolyspora spinosa, was cultured in a variety of media to optimize macrolide titer. Response surface methodology (RSM) was used to improve the fermentation medium and to characterize the microorganism's response to systematic variations in medium composition. Three sequential RSM studies on wild-type A83543 and two high macrolide-producing mutants showed that each strain produced maximum titers in nearly identical fermentation media. No obvious differences in nutrient requirements were evident in the three strains indicating little interaction between mutational change and medium composition through at least two cycles of mutagenesis. The overall increase in macrolide titer starting from the wild-type organism in the original fermentation medium to the second-generation mutant in the optimized medium was over 25-fold.     

Diversity, biomass and distribution pattern of Sargassum beds in the South West lagoon of New Caledonia (South Pacific)

Sargassum (Phaeophyceae, Fucales) is a genus of worldwide distribution recently recognised to proliferate in several regions of the South Pacific. In New Caledonia, species of this genus naturally structure one of the major lagoon habitats but their extent, composition and biomass remain largely unknown. To fill these gaps in our knowledge over large areas, we applied a combination of remote sensing and in situ methods applied to the Neo Caledonian South West Lagoon. Space borne high resolution Landsat (30-m resolution) and Quickbird (2.4-m resolution) images of the Neo Caledonian South West lagoon were used to estimate the spatial extent of the different beds of interest. Species composition was determined for 11 Sargassum beds and seasonal variations were investigated for four representative beds over an 18 month period using quadrats and transects. Relationships between surface cover and biomass were estimated from seasonal field data sets. Finally, four methods requiring variable levels of sampling effort were designed to estimate the total biomass at the scale of each bed, considering (or not) the specific composition, and spatial and temporal variations. Seven Sargassaceae species were identified. Mean surface cover (24.4–51.6%) and total biomass [3.4–1,461.9 t dry matter (DM)] varied widely between beds. Overall, biomass temporal variations were not significantly different, but species-level variations seemed to be bed-specific. The extent of the 11 beds was 9 km²; their total biomass was estimated and compared using each of the four methods, and the most precise method provided an estimate of 2,900 t DM. This study demonstrates a way of characterising Sargassum beds, efficiently and on a large scale, using a combination of remotely sensed and in situ data. These methods should be useful for possible future biomonitoring of Sargassum beds in New Caledonia, and in other areas worldwide.     

Ontogeny of prey attack behaviour in larvae and juveniles of three European cyprinids

Synopsis The ontogenetic change in time costs of prey attacks as well as the change in capture efficiency for representative cladoceran and cyclopoid prey was investigated in roach, Rutilus rutilus, bleak, Alburnus alburnus, and blue bream, Abramis ballerus. Video recordings were used for measuring the timing of attacks, whereas capture efficiencies were determined by direct observation. Decreases in the time cost of attacks reflect the decreasing importance of prey fixation during growth of the fish. No differences in capture efficiencies were found among the three cyprinid species, indicating that attack behaviour is unlikely to function as a basic mechanism leading to differences in prey selectivity among the investigated species. Increasing capture efficiency during early development may lead to increasing selectivity for cyclopoid prey in the field.     

Impaction onto a Glass Slide or Agar versus Impingement into a Liquid for the Collection and Recovery of Airborne Microorganisms

To study impaction versus impingement for the collection and recovery of viable airborne microorganisms, three new bioaerosol samplers have been designed and built. They differ from each other by the medium onto which the bioaerosol particles are collected (glass, agar, and liquid) but have the same inlet and collection geometries and the same sampling flow rate. The bioaerosol concentrations recorded by three different collection techniques have been compared with each other: impaction onto a glass slide, impaction onto an agar medium, and impingement into a liquid. It was found that the particle collection efficiency of agar slide impaction depends on the concentration of agar in the collection medium and on the sampling time, when samples are collected on a nonmoving agar slide. Impingement into a liquid showed anomalous behavior with respect to the sampling flow rate. Optimal sampling conditions in which all three new samplers exhibit the same overall sampling efficiency for nonbiological particles have been established. Inlet and collection efficiencies of about 100% have been achieved for all three devices at a sampling flow rate of 10 liters/min. The new agar slide impactor and the new impinger were then used to study the biological factors affecting the overall sampling efficiency. Laboratory experiments on the total recovery of a typical environmental microorganism, Pseudomonas fluorescens ATCC 13525, showed that both sampling methods, impaction and impingement, provided essentially the same total recovery when relatively nonstressed microorganisms were sampled under optimal sampling conditions. Comparison tests of the newly developed bioaerosol samplers with those commercially available showed that the incorporation of our research findings into the design of the new samplers yields better performance data than data from currently available samplers.     

Estimating breeding season abundance of golden-cheeked warblers in Texas,USA

Population abundance estimates using predictive models are important for describing habitat use and responses to population-level impacts, evaluating conservation status of a species, and for establishing monitoring programs. The golden-cheeked warbler (Setophaga chrysoparia) is a neotropical migratory bird that was listed as federally endangered in 1990 because of threats related to loss and fragmentation of its woodland habitat. Since listing, abundance estimates for the species have mainly relied on localized population studies on public lands and qualitative-based methods. Our goal was to estimate breeding population size of male warblers using a predictive model based on metrics for patches of woodland habitat throughout the species' breeding range. We first conducted occupancy surveys to determine range-wide distribution. We then conducted standard point-count surveys on a subset of the initial sampling locations to estimate density of males. Mean observed patch-specific density was 0.23 males/ha (95% CI = 0.197–0.252, n = 301). We modeled the relationship between patch-specific density of males and woodland patch characteristics (size and landscape composition) and predicted patch occupancy. The probability of patch occupancy, derived from a model that used patch size and landscape composition as predictor variables while addressing effects of spatial relatedness, best predicted patch-specific density. We predicted patch-specific densities as a function of occupancy probability and estimated abundance of male warblers across 63,616 woodland patches accounting for 1.678 million ha of potential warbler habitat. Using a Monte Carlo simulation, our approach yielded a range-wide male warbler population estimate of 263,339 (95% CI: 223,927–302,620). Our results provide the first abundance estimate using habitat and count data from a sampling design focused on range-wide inference. Managers can use the resulting model as a tool to support conservation planning and guide recovery efforts. © 2012 The Wildlife Society.     

Role of the soil Seed Bank in the restoration of a native pinewood at Glen Garry,Inverness-shire

Summary

The potential recovery of ground vegetation in a pinewood at Glen Garry, from which introduced tree species had been removed, was studied by sampling the upper soil seed bank and comparing the species composition of germinating seedlings with extant vegetation. Mean numbers and species of seedlings emerging differed between a control site (not under-planted) and two formerly under-planted sites cleared at different times. Germinating seedling numbers also differed with depth (0–10 cm). Calluna vulgaris and Juncus spp. dominated fifteen species germinating from the seed bank, while some other pinewood species were not found. Although seedling pine occurred, their survival will be affected by competition from Betula spp. and Deschampsia flexuosa. Other elements of the vegetation will recover from the seed bank or by migration, their distribution being influenced by topographic variations and the nature of the developing tree stand. Timing of clearance of exotics in relation to pine seed production and soil scarification might accelerate recovery of the pinewood flora generally.     

Temporal dynamics of gastropod fauna on subtidal sandy sediments of the Ensenada de Baiona (NW Iberian Peninsula)

The temporal variation of the gastropod fauna inhabiting sandy sediments of the Ensenada de Baiona (Galicia, Spain) was studied at three subtidal sites from February 1996 to February 1997 by means of quantitative sampling. A total of 5,463 individuals representing 51 gastropod species and 22 families were found. The family Pyramidellidae was the most diverse in number of species (11 species), followed by Rissoidae and Trochidae (4 species each). The dogwhelk, Nassarius reticulatus, and the rissoid snail, Rissoa parva, were the numerically dominant species at the three studied sites; those and other abundant species showed their greatest densities by the end of summer and the beginning of autumn. In general, univariate measures of the assemblage (number of species, abundance, diversity and evenness) showed variations through time; greater values were recorded between summer and autumn depending on the site. Multivariate analyses done on abundance data showed certain seasonality in the evolution of the assemblage as expected for shallow subtidal sandy sediments at temperate latitudes; those seasonal changes were mostly related to variations in abundance of numerically dominant species. Although the measured sedimentary variables did not show significant correlations with faunal univariate parameters, sediment heterogeneity due to the presence of mats of Zostera marina L. and shells of dead bivalves might explain the differences in composition of the gastropod assemblage among sampling sites.     

Quantum chemical studies on the inhibition potentials of some Penicillin compounds for the corrosion of mild steel in 0.1 M HCl

Inhibitive and adsorption properties of Penicillin G, Amoxicillin and Penicillin V potassium were studied using gravimetric, gasometric and quantum chemical methods. The results obtained indicate that these compounds are good adsorption inhibitors for the corrosion of mild steel in HCl solution. The adsorption of the inhibitors on mild steel surface is spontaneous, exothermic and supports the mechanism of physical adsorption. From DFT results, the sites for nucleophilic attacks in the inhibitors are the carboxylic acid functional group while the sites for electrophilic attacks are in the phenyl ring. There was a strong correlation between theoretical and experimental inhibition efficiencies.     

Comparison of two methods for detection of Giardia cysts and Cryptosporidium oocysts in water.

The steps of two immunofluorescent-antibody-based detection methods were evaluated for their efficiencies in detecting Giardia cysts and Cryptosporidium oocysts. The two methods evaluated were the American Society for Testing and Materials proposed test method for Giardia cysts and Cryptosporidium oocysts in low-turbidity water and a procedure employing sampling by membrane filtration, Percoll-Percoll step gradient, and immunofluorescent staining. The membrane filter sampling method was characterized by higher recovery rates in all three types of waters tested: raw surface water, partially treated water from a flocculation basin, and filtered water. Cyst and oocyst recovery efficiencies decreased with increasing water turbidity regardless of the method used. Recoveries of seeded Giardia cysts exceeded those of Cryptosporidium oocysts in all types of water sampled. The sampling step in both methods resulted in the highest loss of seeded cysts and oocysts. Furthermore, much higher recovery efficiencies were obtained when the flotation step was avoided. The membrane filter method, using smaller tubes for flotation, was less time-consuming and cheaper. A serious disadvantage of this method was the lack of confirmation of presumptive cysts and oocysts, leaving the potential for false-positive Giardia and Cryptosporidium counts when cross-reacting algae are present in water samples.