Microplate fecal coliform method to monitor stream water pollution

A study has been carried out on the Moselle River by means of a microtechnique based on the most-probable-number method for fecal coliform enumeration. This microtechnique, in which each serial dilution of a sample is inoculated into all 96 wells of a microplate, was compared with the standard membrane filter method. It showed a marked overestimation of about 14% due, probably, to the lack of absolute specificity of the method. The high precision of the microtechnique (13%, in terms of the coefficient of variation for log most probable number) and its relative independence from the influence of bacterial density allowed the use of analysis of variance to investigate the effects of spatial and temporal bacterial heterogeneity on the estimation of coliforms. Variability among replicate samples, subsamples, handling, and analytical errors were considered as the major sources of variation in bacterial titration. Variances associated with individual components of the sampling procedure were isolated, and optimal replications of each step were determined. Temporal variation was shown to be more influential than the other three components (most probable number, subsample, sample to sample), which were approximately equal in effect. However, the incidence of sample-to-sample variability (16%, in terms of the coefficient of variation for log most probable number) caused by spatial heterogeneity of bacterial populations in the Moselle River is shown and emphasized. Consequently, we recommend that replicate samples be taken on each occasion when conducting a sampling program for a stream pollution survey.     

Enumeration of cellulolytic anaerobic bacteria from the bovine rumen: Comparison of three methods

Three methods—the most probable number technique, a cellulose agar overlay on basal carbohydrate plates, and carboxymethylcellulose in basal carbohydrate plates—were compared for ease of preparation, interpretation of results, and agreement in estimation of size of the cellulolytic bacterial population in digesta samples from the rumen. The most probable number method yielded consistent detection of cellulose hydrolysis in liquid medium but required at least a 10-day incubation, and its mean was associated with wide 95% confidence intervals. The cellulose overlay method was the least consistent, and zones of hydrolysis often were difficult to see. The carboxymethylcellulose method was the easiest method for preparation and required only a 2-day incubation. The three methods estimated the same population size (all within one-half log unit of each other), but the carboxymethylcellulose method had the lowest coefficient of variation.     

Zooplankton distribution in flowing waters and its implications for sampling: case studies in the River Meuse (Belgium) and the River Moselle (France, Luxembourg)

Zooplankton spatial distribution was studied on four occasions by transect sampling on the rivers Moselle and Meuse in July 1996. Sampling sites were selected for their variety in morphology. A pseudoreplicate-based sampling scheme was adopted that allowed small-scale longitudinal variations in density to be tested along with transversal position and depth. In the Moselle, zooplankton were unevenly distributed transversally, an observation tentatively linked with the complexity of the river channel and the influence of tributaries. In the Meuse, spatial heterogeneity was stronger, and depth also played a key role. Its effect was different for rotifers and microcrustaceans. The occurrence of zooplankton patches, similar to those commonly reported from lakes, was noted. The factors leading to the establishment of such distribution patterns, and their relevance to routine sampling, are briefly discussed; a short list of recommendations for sampling in large rivers is proposed.      

Zooplankton sampling strategies for environmental studies

This study characterizes sources of variation in total zooplankton abundance estimates at seven stations within the 5–10 m depth contour of southeastern Lake Michigan which were sampled monthly, April through October, for the 1975 to 1979 period. Month, year, and station were statistically significant factors affecting abundance estimates as were all interactions. Month was the largest source of variance either as a main effect or interaction. Smallest coefficients of variation were associated with subsampling (mean 6.1%) and replicate sampling (mean 15.1%). The between-station coefficient of variation averaged 39.0% and tended to be highest during the summer. For a given station and month, the between-year coefficient of variation averaged 73.4% while the between-month coefficient of variation for a single station in a given year averaged 95.1%. A table shows the estimated number of replications necessary to detect a true difference in two population means as a function of coefficient of variation. Environmental studies designed to detect spatial alterations should conduct such analyses on a cruise-by-cruise basis. Cruises should consist of a large number of stations and be conducted at least once during each season. Studies designed to detect temporal alterations require more frequent sampling because of the greater variability associated with temporal data sets. Because spatial variability adds little to the overall variability of such data sets, only a few representative stations need be sampled during each cruise.     

Identifying the scales of variability in stream macroinvertebrate abundance, functional composition and assemblage structure

1. Many natural ecosystems are heterogeneous at scales ranging from microhabitats to landscapes. Running waters are no exception in this regard, and their environmental heterogeneity is reflected in the distribution and abundance of stream organisms across multiple spatial scales. 2. We studied patchiness in benthic macroinvertebrate abundance and functional feeding group (FFG) composition at three spatial scales in a boreal river system. Our sampling design incorporated a set of fully nested scales, with three tributaries, two stream sections (orders) within each tributary, three riffles within each section and ten benthic samples in each riffle. 3. According to nested anova s, most of the variation in total macroinvertebrate abundance, abundances of FFGs, and number of taxa was accounted for by the among‐riffle and among‐sample scales. Such small‐scale variability reflected similar patterns of variation in in‐stream variables (moss cover, particle size, current velocity and depth). Scraper abundance, however, varied most at the scale of stream sections, probably mirroring variation in canopy cover. 4. Tributaries and stream sections within tributaries differed significantly in the structure and FFG composition of the macroinvertebrate assemblages. Furthermore, riffles in headwater (second order) sections were more variable than those in higher order (third order) sections. 5. Stream biomonitoring programs should consider this kind of scale‐dependent variability in assemblage characteristics because: (i) small‐scale variability in abundance suggests that a few replicate samples are not enough to capture macroinvertebrate assemblage variability present at a site, and (ii) riffles from the same stream may support widely differing benthic assemblages.     

Distance decay of similarity,effects of environmental noise and ecological heterogeneity among species in the spatio‐temporal dynamics of a dispersal‐limited community

The joint spatial and temporal fluctuations in community structure may be due to dispersal, variation in environmental conditions, ecological heterogeneity among species and demographic stochasticity. These factors are not mutually exclusive, and their relative contribution towards shaping species abundance distributions and in causing species fluctuations have been hard to disentangle. To better understand community dynamics when the exchange of individuals between localities is very low, we studied the dynamics of the freshwater zooplankton communities in 17 lakes located in independent catchment areas, sampled at end of summer from 2002 to 2008 in Norway. We analysed the joint spatial and temporal fluctuations in the community structure by fitting the two‐dimensional Poisson lognormal model under a two‐stage sampling scheme. We partitioned the variance of the distribution of log abundance for a random species at a random time and location into components of demographic stochasticity, ecological heterogeneity among species, and independent environmental noise components for the different species. Non‐neutral mechanisms such as ecological heterogeneity among species (20%) and spatiotemporal variation in the environment (75%) explained the majority of the variance in log abundances. Overdispersion relative to Poisson sampling and demographic stochasticity had a small contribution to the variance (5%). Among a set of environmental variables, lake acidity was the environmental variable that was most strongly related to decay of community similarity in space and time.     

Effect of field sampling design on variation partitioning in a dendritic stream network

Variation partitioning is one of the most frequently used method to infer the importance of environmental (niche based) and spatial (dispersal) processes in metacommunity structuring. However, the reliability of the method in predicting the role of the major structuring forces is less known. We studied the effect of field sampling design on the result of variation partitioning of fish assemblages in a stream network. Along with four different sample sizes, a simple random sampling from a total of 115 stream segments (sampling objects) was applied in 400 iterations, and community variation of each random sample was partitioned into four fractions: pure environmentally (landscape variables) explained, pure spatially (MEM eigenvectors) explained, jointly explained by environment and space, and unexplained variance. Results were highly sensitive to sample size. Even at a given sample size, estimated variance fractions had remarkable random fluctuation, which can lead to inconsistent results on the relative importance of environmental and spatial variables on the structuring of metacommunities. Interestingly, all the four variance fractions correlated better with the number of the selected spatial variables than with any design properties. Sampling interval proved to be a fundamentally influential sampling design property because it affected the number of the selected spatial variables. Our findings suggest that the effect of sampling design on variation partitioning is related to the ability of the eigenvectors to model complex spatial patterns. Hence, properties of the sampling design should be more intensively considered in metacommunity studies.     

Spatial and temporal distribution patterns of drifting pupal exuviae of Chironomidae (Diptera) in streams of tropical northern Australia

1. Periodic collecting of floating cast pupal cuticles of chironomids (exuviae) in two tropical northern Australian streams demonstrates (i) spatial heterogeneity in species composition across a wide stream, (ii) temporal heterogeneity in the maximum abundance of each species, and that (iii) species accumulate as a function of sample size and duration of sampling, 2. Spatial heterogeneity is ascribed to variation in larval microhabitat across the wide stream, combined with short exuvial drift duration and restricted upstream mixing. 3. Temporal heterogeneity is ascribed to diel periodicity in adult emergence and, as with spatial heterogeneity, to the short floating life. 4. The consequences of spatial and temporal variation for the sampling of exuvial drift are discussed in relation to the objectives of particular programmes. Thus, if the objective is assessment of chironomid species composition for inventory purposes such as faunistics or conservation, the large sample sizes attained by 24-h sampling are necessary and appropriate. However, for rapid assessment that requires comparable samples at different sites, species numbers may be optimized by temporally and spatially restricted sampling of the maximal emergence period, which in this study is at dusk, or by subsampling from a 24-h sample.     

Effects of sampling and sub-sampling variation using the STAR-AQEM sampling protocol on the precision of macroinvertebrate metrics

As part of the extensive field sampling programme within the European Union STAR project, replicate macroinvertebrate samples were taken using the STAR-AQEM sampling method at each of 2–13 sites of varying ecological quality within each of 15 stream types spread over 12 countries throughout Europe. The STAR-AQEM method requires the sub-sampling and taxonomic identification of at least one-sixth of the sample and at least 700 individuals. Replicate sub-samples were also taken at most of these sites. Sub-sampling effects caused more than 50% of the overall variance between replicate samples values for 12 of the 27 macroinvertebrate metrics analysed and was generally greatest for metrics that depend on the number of taxa present. The sampling precision of each metric was estimated by the overall replicate sampling variance as a percentage P_samp of the total variance in metric values within a stream type. Average over all stream types, the three Saprobic indices had the lowest percentage sampling variances with median values of only 3–6%. Most of the metrics had typical replicate sampling variances of 8–18% of the total variability within a stream type; this gives rise to estimated rates of mis-classifying sites to ecological status class of between 22 and 55% with an average of about 40%. This suggests that the precision of such metrics based on the STAR-AQEM method is only sufficient to indicate gross changes in the ecological status of sites, but there will be considerable uncertainty in the assignment of sites to adjacent status classes. These estimates can be used to provide information on the effects of STAR-AQEM sampling variation on the expected uncertainty in multi-metric assessments of the ecological status of sites in the same or similar stream types, where only one sample has been taken at a point in time and thus there is no replication.     

Insect herbivores associated with Baccharis dracunculifolia (Asteraceae): responses of gall-forming and free-feeding insects to latitudinal variation

The spatial heterogeneity hypothesis has been invoked to explain the increase in species diversity from the poles to the tropics: the tropics may be more diverse because they contain more habitats and micro-habitats. In this paper, the spatial heterogeneity hypothesis prediction was tested by evaluating the variation in richness of two guilds of insect herbivores (gall-formers and free-feeders) associated with Baccharis dracunculifolia (Asteraceae) along a latitudinal variation in Brazil. The seventeen populations of B. dracunculifolia selected for insect herbivores sampling were within structurally similar habitats, along the N-S distributional limit of the host plant, near the Brazilian sea coast. Thirty shrubs were surveyed in each host plant population. A total of 8 201 galls and 864 free-feeding insect herbivores belonging to 28 families and 88 species were sampled. The majority of the insects found on B. dracunculifolia were restricted to a specific site rather than having a geographic distribution mirroring that of the host plant. Species richness of free-feeding insects was not affected by latitudinal variation corroborating the spatial heterogeneity hypothesis. Species richness of gall-forming insects was positively correlated with latitude, probably because galling insect associated with Baccharris genus radiated in Southern Brazil. Other diversity indices and evenness estimated for both gall-forming and free feeding insect herbivores, did not change with latitude, suggesting a general structure for different assemblages of herbivores associated with the host plant B. dracunculifolia. Thus it is probable that, insect fauna sample in each site resulted of large scale events, as speciation, migration and coevolution, while at local level, the population of these insects is regulated by ecological forces which operate in the system.     

Macroinvertebrate community structure along gradients of hydraulic and sedimentary conditions in a large gravel-bed river

1. The spatial distribution of macroinvertebrate species was examined in relation to hydraulic and sedimentary conditions in a large gravel‐bed river, the Fraser River, Canada. Mean annual discharge in the Fraser River is 2900 m3 s?1 and annual flood discharge, due to snowmelt in May and June, averages 8760 m3 s?1.
2. Invertebrates were sampled from four water depths (0.2, 0.5, 1.5, 3.0 m) at various levels of discharge that together captured the spatial and temporal variability of the physical habitat. Several hydraulic (near‐bed shear velocity, Boundary Reynolds number, turbulence intensity, depth‐averaged velocity, Froude number, Reynolds number) and substratum variables (mean grain size, Trask's sorting coefficient, Nikuradse's roughness, percentage of fine sediment, and Shields entrainment function) were measured for each sample of macroinvertebrates. Concentrations of fine and coarse particulate organic matter were also assessed.
3. The physical habitat was characterized by a major gradient of hydraulic conditions that corresponded positively with increasing water depth and accounted for 52% of the total variation in the habitat data. Substratum conditions and the concentration of organic matter explained 24% of the total variation in the habitat data.
4. The distribution of invertebrates was correlated significantly with hydraulic variables and suggests that hydraulic conditions represent a major physical gradient along which the benthic community is organized. The distribution of organic matter and substratum texture were also important for some species. The spatial distribution of most species reflected morphological and trophic suitability to particular habitat conditions.
5. Hydraulic stress associated with foraging and maintaining position, as well as organic matter retention in coarse substrata, are probable mechanisms affecting the spatial distribution of macroinvertebrates.     

Apolipoprotein B signal peptide polymorphism distribution among south Amerindian populations

We report the distribution of the APOB signal peptide polymorphism in 5 native populations of South America: 2 samples of Mataco and 1 sample each of Pilagá and Toba from the Argentinian Chaco and 1 sample of Ache from the Paraguay forest. A randomly selected subsample of a previously studied sample from the Cayapa of Ecuador (Scacchi et al. 1997) was reanalyzed to investigate probable differences attributable to sampling, laboratory techniques, or interobserver error. The polymorphism observed in the signal peptide region of the APOB gene among native populations of South America exhibits the same range of variation found among geographic continental populations, confirming the high genetic heterogeneity of South Amerindians. Extremes in the allele prevalences were found among the Mataco and Ache, populations not far apart geographically. The small differences in genotype and allele frequencies between the subsample of the Cayapa analyzed here and the original Cayapa sample and between the 2 Mataco samples were not statistically significant and most likely were due to sampling error.     

Fine-scale spatial patterns in bacterial community composition and function within freshwater ponds

The extent to which non-host-associated bacterial communities exhibit small-scale biogeographic patterns in their distribution remains unclear. Our investigation of biogeography in bacterial community composition and function compared samples collected across a smaller spatial scale than most previous studies conducted in freshwater. Using a grid-based sampling design, we abstracted 100+ samples located between 3.5 and 60 m apart within each of three alpine ponds. For every sample, variability in bacterial community composition was monitored using a DNA-fingerprinting methodology (automated ribosomal intergenic spacer analysis) whereas differences in bacterial community function (that is, carbon substrate utilisation patterns) were recorded from Biolog Ecoplates. The exact spatial position and dominant physicochemical conditions (for example, pH and temperature) were simultaneously recorded for each sample location. We assessed spatial differences in bacterial community composition and function within each pond and found that, on average, community composition or function differed significantly when comparing samples located >20 m apart within any pond. Variance partitioning revealed that purely spatial variation accounted for more of the observed variability in both bacterial community composition and function (range: 24–38% and 17–39%) than the combination of purely environmental variation and spatially structured environmental variation (range: 17–32% and 15–20%). Clear spatial patterns in bacterial community composition, but not function were observed within ponds. We therefore suggest that some of the observed variation in bacterial community composition is functionally ‘redundant''. We confirm that distinct bacterial communities are present across unexpectedly small spatial scales suggesting that populations separated by distances of >20 m may be dispersal limited, even within the highly continuous environment of lentic water.     

Pulmonary blood flow remains fractal down to the level of gas exchange.

The spatial distribution of pulmonary blood flow is increasingly heterogeneous as progressively smaller lung regions are examined. To determine the extent of perfusion heterogeneity at the level of gas exchange, we studied blood flow distributions in rat lungs by using an imaging cryomicrotome. Approximately 150,000 fluorescent 15-microm-diameter microspheres were injected into tail veins of five awake rats. The rats were heavily anesthetized; the lungs were removed, filled with an optimal cutting tissue compound, and frozen; and the spatial location of every microsphere was determined. The data were mathematically dissected with the use of an unbiased random sampling method. The coefficients of variation of microsphere distributions were determined at varying sampling volumes. Perfusion heterogeneity increased linearly on a log-log plot of coefficient of variation vs. volume, down to the smallest sampling size of 0.53 mm(3). The average fractal dimension, a scale-independent measure of perfusion distribution, was 1.2. This value is similar to that of other larger species such as dogs, pigs, and horses. Pulmonary perfusion heterogeneity increases continuously and remains fractal down to the acinar level. Despite the large degree of perfusion heterogeneity at the acinar level, gases are efficiently exchanged.     

两个林分水曲柳土壤种子库空间格局的定量比较

依据地统计理论和方法,采取规则网格数据结构的空间取样设计,在树种组成、林分密度及林下植被等均不相同的两个林分中设置样地,对水曲柳土壤种子库的空间格局进行了定量比较研究．结果表明,在水曲柳杨树等混生的样地1,种子库密度较大(158．1±96．7个·m^-2),异质性程度较高(C0+C＝0．549),格局尺度相对较大(4．52m),空间自相关因素引起的变异高达89．4％,随机变异仅占10．6％,格局变异呈现较高的自相似性(分数维值D＝1．728-1．865)．在水曲柳纯林中的样地2,种子库密度较小(57．4±44．7个·m^-2),异质性程度较低(C0+C＝0．125),且主要体现在相对较小的尺度范围内(2．59m),空间自相关变异占56．8％,随机因素对空间格局的影响较大(D＝1．906-1．960)．两样地之间种子库空间格局表现的尺度范围、强度及空间结构等有显著差异,林分空间异质性是导致这种差异的主要原因．     

The quantification of local substrate heterogeneity in streams and its significance for macroinvertebrate assemblages

The effect of substrate heterogeneity on the structure of stream macroinvertebrate assemblages (total abundance, taxon richness, and evenness) is still not clear, but this could be due to the lack of standard methods for quantifying substrate heterogeneity. An accurate quantification of substrate heterogeneity was obtained from photographs of sampled areas (each 225 cm²), which were used to create maps that were subsequently digitized and analyzed using image analysis software. These maps allowed the calculation of multiple metrics quantifying two aspects of substrate heterogeneity: composition and spatial configuration of substrate patches. The diversity of substrate types (calculated as the Shannon diversity index), and the heterogeneity of patch compactness (calculated as the coefficient of variation of the relationship between patch dimensions) were the metrics explaining more biotic variance at the sample scale, but at higher scales there were no relationships between assemblage structure and substrate heterogeneity. Most variation in substrate heterogeneity occurred at the sample scale, while some metrics varied significantly at riffle or segment scales; these patterns of variation match those of macroinvertebrate assemblages, which had been previously studied. The importance of quantifying substrate heterogeneity and considering the spatial scales of its study are discussed.     

Assessment of species diversity from species abundance distributions at different localities

We show how the spatial structure of species diversity can be analyzed using the correlation between the log abundances of the species in the communities, assuming that two communities at different localities can be described by a bivariate lognormal species abundance distribution. A useful property of this approach is that the log abundances of the species at two localities can be considered as samples from a bivariate normal distribution defined by only five parameters. The variances and the correlation can be estimated by maximum likelihood methods even if there is no information about the sampling intensity and the number of unobserved species. This method also enables estimation of over-dispersion in the sampling relative to a Poisson distribution that allows sampling adjustment of the estimate of β-diversity. Furthermore, we also obtain a partitioning of species diversity into additive components of α-, β- and γ-diversity. For instance, if the correlation between the log abundances of the species is close to one, the same species will be common and rare in the two communities and the β-diversity will be low. We illustrate this approach by analysing similarities of communities of rare and endangered species of oak-living beetles in south-eastern Norway. The number of recorded species was estimated to be only 48.1% of the total number of species actually present in these communities. The correlations among communities dropped rather quickly with distance with a scaling of order 200 km. This illustrates large spatial heterogeneity in species composition, which should be accounted for in the design of schemes of such devices for assessing species diversity in these habitat-types.     

Power, precaution, Type II error and sampling design in assessment of environmental impacts

Increasingly, environmental managers attempt to incorporate precautionary principles into decision making. In any quantitative analysis of impacts, precaution is closely related to the power of the analysis to detect an impact. Designs of sampling to detect impacts are, however, complex because of natural spatial and temporal variability and the intrinsic nature of the statistical interactions which define impacts. Here, pulse and press responses and impacts that affect time courses (temporal variance) were modelled to determine the influences of increasing temporal replication—sampling more times in each of several longer periods before and again after an impact.Increasing the number of control or reference locations and number of replicate sample units at each time and place of sampling investigated the influence of spatial replication on power. From numerous scenarios of impacts, with or without natural spatial and temporal interactions (i.e. not caused by an impact), general recommendations are possible. Detecting press impacts requires maximal numbers of control locations. Shorter-term pulse impacts are best detected when the number of periods sampled is maximized. Impacts causing changes in temporal variance are most likely to be detected by sampling with the greatest possible number of periods or times within periods.To allow precautionary decision making, the type of predicted impact should be specified with its magnitude and duration. Only then can sampling be designed to be powerful, thereby allowing precautionary concepts to be invoked.