Quantifying Change Through Time in Spider Assemblages: Sampling Methods, Indices and Sources of Error

Using a sample dataset from six sites in the Southern Appalachian mountains in the United States, I evaluate the usefulness of diversity indices and similarity/difference indices for monitoring the changes in spider (Order Araneae) assemblages through time. The Shannon index, Brillouin index, Simpson's index and Margalef's index were correlated with each other and were able to detect successional changes in two old clear-cuts as well as disturbance to a Beech gap forest possibly due to Beech Bark Disease. Turnover, Bray-Curtis similarity, and Morisita-Horn similarity also detected successional differences between the forest types and indicated the short-term and long-term changes in a mature forest stand. Seasonal changes (early to late summer) in spider communities were consistently higher than yearly changes. I identify and discuss the implications of various sources of error that will adversely affect the accuracy and efficiency of a monitoring protocol: sensitivity of indices to sample size, sampler bias, inclusion of juveniles, taxonomic uncertainty and stochastic sampling effects. Although gross trends are still apparent in the sample data regardless of these errors, more subtle changes may be obscured. It is these subtle trends which may be more useful to managers attempting to identify disturbance before irreversible damage occurs.     

Field Evaluation of Personal Sampling Methods for Multiple Bioaerosols

Ambient bioaerosols are ubiquitous in the daily environment and can affect health in various ways. However, few studies have been conducted to comprehensively evaluate personal bioaerosol exposure in occupational and indoor environments because of the complex composition of bioaerosols and the lack of standardized sampling/analysis methods. We conducted a study to determine the most efficient collection/analysis method for the personal exposure assessment of multiple bioaerosols. The sampling efficiencies of three filters and four samplers were compared. According to our results, polycarbonate (PC) filters had the highest relative efficiency, particularly for bacteria. Side-by-side sampling was conducted to evaluate the three filter samplers (with PC filters) and the NIOSH Personal Bioaerosol Cyclone Sampler. According to the results, the Button Aerosol Sampler and the IOM Inhalable Dust Sampler had the highest relative efficiencies for fungi and bacteria, followed by the NIOSH sampler. Personal sampling was performed in a pig farm to assess occupational bioaerosol exposure and to evaluate the sampling/analysis methods. The Button and IOM samplers yielded a similar performance for personal bioaerosol sampling at the pig farm. However, the Button sampler is more likely to be clogged at high airborne dust concentrations because of its higher flow rate (4 L/min). Therefore, the IOM sampler is a more appropriate choice for performing personal sampling in environments with high dust levels. In summary, the Button and IOM samplers with PC filters are efficient sampling/analysis methods for the personal exposure assessment of multiple bioaerosols.     

The Robustness of Plant-Pollinator Assemblages: Linking Plant Interaction Patterns and Sensitivity to Pollinator Loss

Most flowering plants depend on pollinators to reproduce. Thus, evaluating the robustness of plant-pollinator assemblages to species loss is a major concern. How species interaction patterns are related to species sensitivity to partner loss may influence the robustness of plant-pollinator assemblages. In plants, both reproductive dependence on pollinators (breeding system) and dispersal ability may modulate plant sensitivity to pollinator loss. For instance, species with strong dependence (e.g. dioecious species) and low dispersal (e.g. seeds dispersed by gravity) may be the most sensitive to pollinator loss. We compared the interaction patterns of plants differing in dependence on pollinators and dispersal ability in a meta-dataset comprising 192 plant species from 13 plant-pollinator networks. In addition, network robustness was compared under different scenarios representing sequences of plant extinctions associated with plant sensitivity to pollinator loss. Species with different dependence on pollinators and dispersal ability showed similar levels of generalization. Although plants with low dispersal ability interacted with more generalized pollinators, low-dispersal plants with strong dependence on pollinators (i.e. the most sensitive to pollinator loss) interacted with more particular sets of pollinators (i.e. shared a low proportion of pollinators with other plants). Only two assemblages showed lower robustness under the scenario considering plant generalization, dependence on pollinators and dispersal ability than under the scenario where extinction sequences only depended on plant generalization (i.e. where higher generalization level was associated with lower probability of extinction). Overall, our results support the idea that species generalization and network topology may be good predictors of assemblage robustness to species loss, independently of plant dispersal ability and breeding system. In contrast, since ecological specialization among partners may increase the probability of disruption of interactions, the fact that the plants most sensitive to pollinator loss interacted with more particular pollinator assemblages suggest that the persistence of these plants and their pollinators might be highly compromised.     

Antimicrobial Histones and DNA Traps in Invertebrate Immunity: EVIDENCES IN CRASSOSTREA GIGAS*

Although antimicrobial histones have been isolated from multiple metazoan species, their role in host defense has long remained unanswered. We found here that the hemocytes of the oyster Crassostrea gigas release antimicrobial H1-like and H5-like histones in response to tissue damage and infection. These antimicrobial histones were shown to be associated with extracellular DNA networks released by hemocytes, the circulating immune cells of invertebrates, in response to immune challenge. The hemocyte-released DNA was found to surround and entangle vibrios. This defense mechanism is reminiscent of the neutrophil extracellular traps (ETs) recently described in vertebrates. Importantly, oyster ETs were evidenced in vivo in hemocyte-infiltrated interstitial tissues surrounding wounds, whereas they were absent from tissues of unchallenged oysters. Consistently, antimicrobial histones were found to accumulate in oyster tissues following injury or infection with vibrios. Finally, oyster ET formation was highly dependent on the production of reactive oxygen species by hemocytes. This shows that ET formation relies on common cellular and molecular mechanisms from vertebrates to invertebrates. Altogether, our data reveal that ET formation is a defense mechanism triggered by infection and tissue damage, which is shared by relatively distant species suggesting either evolutionary conservation or convergent evolution within Bilateria.     

Evaluation of Four Aerobiological Sampling Methods for the Retrieval of Aerosolized Pseudomonas syringae

[This corrects the article on p. 1268 in vol. 57.].     

Evaluation of Four Aerobiological Sampling Methods for the Retrieval of Aerosolized Pseudomonas syringae

The Andersen six-stage impactor, the SAS (Surface Air System) impactor, the AGI-30 impinger, and gravity plates were evaluated for the retrieval of aerosol-released Pseudomonas syringae. The upper limits of the impactor samplers were exceeded at a spray concentration of 10⁷ CFU/ml, indicating that these samplers are not appropriate for monitoring high airborne concentrations. Decreased cell concentrations were retrieved with increased sampling time for the Andersen and AGI samplers, indicating that a minimum sampling time is preferable for monitoring aerosolized vegetative cells.     

Methods for Sampling of Airborne Viruses

Summary: To better understand the underlying mechanisms of aerovirology, accurate sampling of airborne viruses is fundamental. The sampling instruments commonly used in aerobiology have also been used to recover viruses suspended in the air. We reviewed over 100 papers to evaluate the methods currently used for viral aerosol sampling. Differentiating infections caused by direct contact from those caused by airborne dissemination can be a very demanding task given the wide variety of sources of viral aerosols. While epidemiological data can help to determine the source of the contamination, direct data obtained from air samples can provide very useful information for risk assessment purposes. Many types of samplers have been used over the years, including liquid impingers, solid impactors, filters, electrostatic precipitators, and many others. The efficiencies of these samplers depend on a variety of environmental and methodological factors that can affect the integrity of the virus structure. The aerodynamic size distribution of the aerosol also has a direct effect on sampler efficiency. Viral aerosols can be studied under controlled laboratory conditions, using biological or nonbiological tracers and surrogate viruses, which are also discussed in this review. Lastly, general recommendations are made regarding future studies on the sampling of airborne viruses.     

Ecosystem Linkages between Southern Appalachian Headwater Streams and Their Banks: Leaf Litter Breakdown and Invertebrate Assemblages

We examined red maple (Acer rubrum L.) leaf litter breakdown in streams and riparian zones at two sites in the southern Appalachian Mountains to understand how differences in abiotic and biotic factors influence leaf breakdown rates. Litterbags were placed in three riparian habitats differing in litter layer moisture: stream > bank > upland. Invertebrates colonizing litterbags at one site were also examined to determine how variations in community and functional structure affect breakdown rates. Leaves broke down fastest in streams and slowest in upland habitats, whereas bank habitats were intermediate and characterized by high variability. Faster leaf breakdown rates in streams appeared to be a function of greater moisture availability, a more stable thermal regime, and a higher biomass of leaf-shredding invertebrates, especially the stonefly Tallaperla. In addition, patterns of leaf breakdown and invertebrate community structure provided evidence for a stronger than expected ecological connection between the stream and the bank. Overall, detritus processing within this narrow riparian ecosystem varied considerably depending on the availability of moisture. Results from this study show that stream channel–floodplain interactions in riparian ecosystems of steep forested mountains are analogous to ones in larger downstream or low-gradient systems. Riparian zones throughout a river network display a remarkable heterogeneity in their ability to process organic matter, which is ultimately driven by changes in hydrological conditions. Received 6 March 2001; accepted 3 July 2001.     

Marine Invertebrate Conservation: Evaluation of Fisheries Over-exploitation Concerns

The nature of overfishing of marine invertebrates is complex,ranging from the perception of overfishing because of competitionby user groups for a common property resource to extensive overfishingto near extinction because of poaching by either licensed orunlicensed fishers. As a group, marine invertebrates seem particularlyresistant to overfishing, primarily because their relative immobilityand scattered concentrations means refuge populations oftenexist. However, this distribution pattern also means a fisheryis scattered over an often large geographical area with relativelysmall, frequent landings at any location. A minimum legal sizeregulation, enforceable anywhere before consumption, is theprimary regulation applied by managers for many species to ensureagainst overfishing. Overfishing concerns arise primarily forthose species where price is sufficiently high to encourageillegal fishing or where harvest by fishers is not easily monitoredor controlled because of the nature of the fishing activityor because only part of the animal (e.g., the flesh) is harvested.Instances of overfishing by cause are discussed, and examplesare presented to demonstrate how managers are dealing with orhave dealt with different situations.     

Searching for the Optimal Sampling Solution: Variation in Invertebrate Communities,Sample Condition and DNA Quality

There is a great demand for standardising biodiversity assessments in order to allow optimal comparison across research groups. For invertebrates, pitfall or flight-interception traps are commonly used, but sampling solution differs widely between studies, which could influence the communities collected and affect sample processing (morphological or genetic). We assessed arthropod communities with flight-interception traps using three commonly used sampling solutions across two forest types and two vertical strata. We first considered the effect of sampling solution and its interaction with forest type, vertical stratum, and position of sampling jar at the trap on sample condition and community composition. We found that samples collected in copper sulphate were more mouldy and fragmented relative to other solutions which might impair morphological identification, but condition depended on forest type, trap type and the position of the jar. Community composition, based on order-level identification, did not differ across sampling solutions and only varied with forest type and vertical stratum. Species richness and species-level community composition, however, differed greatly among sampling solutions. Renner solution was highly attractant for beetles and repellent for true bugs. Secondly, we tested whether sampling solution affects subsequent molecular analyses and found that DNA barcoding success was species-specific. Samples from copper sulphate produced the fewest successful DNA sequences for genetic identification, and since DNA yield or quality was not particularly reduced in these samples additional interactions between the solution and DNA must also be occurring. Our results show that the choice of sampling solution should be an important consideration in biodiversity studies. Due to the potential bias towards or against certain species by Ethanol-containing sampling solution we suggest ethylene glycol as a suitable sampling solution when genetic analysis tools are to be used and copper sulphate when focusing on morphological species identification and facing financial restrictions in biodiversity studies.     

急性肺栓塞常用检查方法及效果评价

肺动脉栓塞的诊断虽在临床工作中较少见到,但却是一种常见的致死性急症,却易被广大临床医务工作者所忽视。本文综述几种简单有效的检查方法,以提高急性PE的诊断率及便于及早治疗并降该病的病死率,并为进一步防治肺动脉栓塞提供参考依据。     

Synthesis and Evaluation of DMPO-Type Spin Traps

The spin traps substituted with some groups at the 4-position of dimethyl-1-pyrroline N-oxide(DMPO) were compared with DMPO itself regarding their abilities as spin traps and their physical properties. 4,5,5-Trimethyl-l-pyroHine N-oxide (4MDMPO) and 5,5-dimethyl- 4-phenyl-l-pyrolline N-oxide (4PDMPO) were synthesized by the Bonnett method, and 5,5-dimethyl-4-hydroxymethyl-l-pyrolline N-oxide (4HMDMPO) was made by a unique method from 2(5H)-furanone. The melting points of 4MDMPO, 4PDMPO and 4HMDMPO were higher than that of DMPO. The magnitude of hydrophilicity was in the order of 4HMDMPO, DMPO, 4MDMPO, and 4PDMPO based on the partition coefficient experiments in a 1-octanol-water system. Several radicals, O₂, HO-, -CH₃, -CH₂OH, -CH(CH₃)OH, (CH₃)₃ CO and H radicals, were trapped with these DMPO derivatives for comparison with the trapping by DMPO itself. Spin adducts of O J with the three DMPO derivatives showed ESR spectra similar to that of DMPO. In spite of the formation of diastereomers arising from spin trapping, the line-width enlargement was very small. The intensities and the decay rates of the spectra of 4MDMPO-O₂^-, 4PDMPO-O₂^- 4HMDMPO-O₂^- and DMPO-O₂^- were almost equal. In the trapping of the OH radical by 4MDMPO, 4PDMPO and 4HMDMPO, the eight-line ESR spectra observed were different from the well-known four-line spectrum of DMPO-OH.     

Evaluation of Methods for Sampling, Recovery, and Enumeration of Bacteria Applied to the Phylloplane

Determining the fate and survival of genetically engineered microorganisms released into the environment requires the development and application of accurate and practical methods of detection and enumeration. Several experiments were performed to examine quantitative recovery methods that are commonly used or that have potential applications. In these experiments, Erwinia herbicola and Enterobacter cloacae were applied in greenhouses to Blue Lake bush beans (Phaseolus vulgaris) and Cayuse oats (Avena sativa). Sampling indicated that the variance in bacterial counts among leaves increased over time and that this increase caused an overestimation of the mean population size by bulk leaf samples relative to single leaf samples. An increase in the number of leaves in a bulk sample, above a minimum number, did not significantly reduce the variance between samples. Experiments evaluating recovery methods demonstrated that recovery of bacteria from leaves was significantly better with stomacher blending, than with blending, sonication, or washing and that the recovery efficiency was constant over a range of sample inoculum densities. Delayed processing of leaf samples, by storage in a freezer, did not significantly lower survival and recovery of microorganisms when storage was short term and leaves were not stored in buffer. The drop plate technique for enumeration of bacteria did not significantly differ from the spread plate method. Results of these sampling, recovery, and enumeration experiments indicate a need for increased development and standardization of methods used by researchers as there are significant differences among, and also important limitations to, some of the methods used.     

Fish Assemblages Associated with Natural and Anthropogenically-Modified Habitats in a Marine Embayment: Comparison of Baited Videos and Opera-House Traps

Marine embayments and estuaries play an important role in the ecology and life history of many fish species. Cockburn Sound is one of a relative paucity of marine embayments on the west coast of Australia. Its sheltered waters and close proximity to a capital city have resulted in anthropogenic intrusion and extensive seascape modification. This study aimed to compare the sampling efficiencies of baited videos and fish traps in determining the relative abundance and diversity of temperate demersal fish species associated with naturally occurring (seagrass, limestone outcrops and soft sediment) and modified (rockwall and dredge channel) habitats in Cockburn Sound. Baited videos sampled a greater range of species in higher total and mean abundances than fish traps. This larger amount of data collected by baited videos allowed for greater discrimination of fish assemblages between habitats. The markedly higher diversity and abundances of fish associated with seagrass and limestone outcrops, and the fact that these habitats are very limited within Cockburn Sound, suggests they play an important role in the fish ecology of this embayment. Fish assemblages associated with modified habitats comprised a subset of species in lower abundances when compared to natural habitats with similar physical characteristics. This suggests modified habitats may not have provided the necessary resource requirements (e.g. shelter and/or diet) for some species, resulting in alterations to the natural trophic structure and interspecific interactions. Baited videos provided a more efficient and non-extractive method for comparing fish assemblages and habitat associations of smaller bodied species and juveniles in a turbid environment.     

Sampling Plan Optimization: A Data Review and Sampling Frequency Evaluation Process

Lawrence Livermore National Laboratory (LLNL) uses a cost-effective sampling (CES) methodology to evaluate and review ground water contaminant data and optimize the site's ground water monitoring plan. The CES methodology is part of LLNL's regulatory approved compliance monitoring plan (Lamarre et al., 1996). It allows LLNL to adjust the ground water sampling plan every quarter in response to changing conditions at the site. Since the use of the CES methodology has been approved by the appropriate regulatory agencies, such adjustments do not need additional regulatory approval. This permits LLNL to respond more quickly to changing conditions. The CES methodology bases the sampling frequency for each location on trend, variability, and magnitude statistics describing the contaminants at that location, and on the input of the technical staff (hydrologists, chemists, statisticians, and project leaders). After initial setup is complete, each application of CES takes only a few days for as many as 400 wells. Effective use of the CES methodology requires sufficient data, an understanding of contaminant transport at the site, and an adequate number of monitoring wells downgradient of the contamination. The initial implementation of CES at LLNL in 1992 produced a 40% reduction in the required number of annual routine ground water samples at LLNL. This has saved LLNL $390,000 annually in sampling, analysis, and data management costs.     

Evaluation of the Biological Sampling Kit (BiSKit) for Large-Area Surface Sampling

Current surface sampling methods for microbial contaminants are designed to sample small areas and utilize culture analysis. The total number of microbes recovered is low because a small area is sampled, making detection of a potential pathogen more difficult. Furthermore, sampling of small areas requires a greater number of samples to be collected, which delays the reporting of results, taxes laboratory resources and staffing, and increases analysis costs. A new biological surface sampling method, the Biological Sampling Kit (BiSKit), designed to sample large areas and to be compatible with testing with a variety of technologies, including PCR and immunoassay, was evaluated and compared to other surface sampling strategies. In experimental room trials, wood laminate and metal surfaces were contaminated by aerosolization of Bacillus atrophaeus spores, a simulant for Bacillus anthracis, into the room, followed by settling of the spores onto the test surfaces. The surfaces were sampled with the BiSKit, a cotton-based swab, and a foam-based swab. Samples were analyzed by culturing, quantitative PCR, and immunological assays. The results showed that the large surface area (1 m²) sampled with the BiSKit resulted in concentrations of B. atrophaeus in samples that were up to 10-fold higher than the concentrations obtained with the other methods tested. A comparison of wet and dry sampling with the BiSKit indicated that dry sampling was more efficient (efficiency, 18.4%) than wet sampling (efficiency, 11.3%). The sensitivities of detection of B. atrophaeus on metal surfaces were 42 ± 5.8 CFU/m² for wet sampling and 100.5 ± 10.2 CFU/m² for dry sampling. These results demonstrate that the use of a sampling device capable of sampling larger areas results in higher sensitivity than that obtained with currently available methods and has the advantage of sampling larger areas, thus requiring collection of fewer samples per site.