Influence of geographical extent on typology- and model-based assessments of taxonomic completeness of river macroinvertebrates

1. River InVertebrate Prediction and Classification System (RIVPACS)‐type predictive models are increasingly used to assess the biological condition of freshwaters, but management schemes may also be based on a priori groupings of similar water bodies (typologies) to control for natural variation in biota. The two approaches may lead to disagreements of the biological status of a site, depending on, for example, the spatial scale at which assessments are conducted. 2. We used data from 96 reference and 134 potentially impacted sites from Western and Central Finland to compare RIVPACS‐type models and a simple size‐based typology of rivers for the assessment of taxonomic completeness (the quotient of the Observed‐to‐Expected number of predicted taxa, O / E) of riffle macroinvertebrates. We specifically examined how geographical extent influences bioassessment performance (accuracy, precision and sensitivity to detect impact) of the two approaches. To fully examine the behaviour of the O / E‐index with the two approaches at differing spatial scales, we performed all assessments with a full range of thresholds for predicted taxa occurrence probabilities (p_t from 0+ to 0.9). 3. Both approaches performed consistently better than the corresponding null models. At the larger extent (i.e. assessment encompassing the whole study area), the RIVPACS‐approach performed in all aspects better than the typology‐approach. However, at the smaller extent (i.e. regional assessments) the RIVPACS‐type models and the typologies showed similar accuracy to predict the actual fauna (mean E), similar precision (SD) of cross‐validated O / E and similar sensitivity to detect sites with human impairment. 4. SD(O / E) decreased (i.e. precision increased) consistently with increasing p_t. However, both approaches were most sensitive at intermediate p_t:s (c. 0.2–0.6) when taxa with low predicted occurrence probabilities were excluded. 5. Our results show that RIVPACS‐type predictive models are less susceptible to variations in spatial scale, whereas the performance of a priori typologies increases with decreasing spatial extent. Thus, RIVPACS‐type models are more efficient for large‐scale bioassessments, but at restricted spatial scales, or with an otherwise biologically meaningful stratification, simple a priori classifications can be equally useful for the assessment of taxonomic completeness of river macroinvertebrates.     

ANNA: A new prediction method for bioassessment programs

1. Cluster analysis of reference sites with similar biota is the initial step in creating River Invertebrate Prediction and Classification System (RIVPACS) and similar river bioassessment models such as Australian River Assessment System (AUSRIVAS). This paper describes and tests an alternative prediction method, Assessment by Nearest Neighbour Analysis (ANNA), based on the same philosophy as RIVPACS and AUSRIVAS but without the grouping step that some people view as artificial. 2. The steps in creating ANNA models are: (i) weighting the predictor variables using a multivariate approach analogous to principal axis correlations, (ii) calculating the weighted Euclidian distance from a test site to the reference sites based on the environmental predictors, (iii) predicting the faunal composition based on the nearest reference sites and (iv) calculating an observed/expected (O/E) analogous to RIVPACS/AUSRIVAS. 3. The paper compares AUSRIVAS and ANNA models on 17 datasets representing a variety of habitats and seasons. First, it examines each model's regressions for Observed versus Expected number of taxa, including the r², intercept and slope. Second, the two models’ assessments of 79 test sites in New Zealand are compared. Third, the models are compared on test and presumed reference sites along a known trace metal gradient. Fourth, ANNA models are evaluated for western Australia, a geographically distinct region of Australia. The comparisons demonstrate that ANNA and AUSRIVAS are generally equivalent in performance, although ANNA turns out to be potentially more robust for the O versus E regressions and is potentially more accurate on the trace metal gradient sites. 4. The ANNA method is recommended for use in bioassessment of rivers, at least for corroborating the results of the well established AUSRIVAS‐ and RIVPACS‐type models, if not to replace them.     

Evaluation of approaches for measuring taxonomic completeness of lake profundal macroinvertebrate assemblages

1. Benthic macroinvertebrates (MI) are commonly used to assess freshwater ecosystems with the reference condition approach. Such assessments necessitate control for natural community variation, either by categorical typologies or by predictive models that have been widely and successfully developed for running water biota but not previously for lake profundal invertebrates. 2. We evaluated four modelling techniques [multivariate regression tree (MRT), limiting environmental differences, nonparametric multiplicative regression (NPMR) and River Invertebrate Prediction And Classification System (RIVPACS) and the operative Finnish lake typology for assessing taxonomic completeness (observed‐to‐expected number of taxa, O/E) of profundal MI assemblages. We used data from 74 and 33 minimally disturbed reference lake basins for calibration and validation of the approaches, respectively, and 72 test basins subject to various anthropogenic pressures to evaluate sensitivity to detect impact. Either all predicted taxa (threshold probability of capture P_t = 0+) or only those predicted to be captured with ≥0.25 probability were used to calculate O/E. 3. With P_t = 0.25, all four modelling approaches were accurate (mean O/E = 0.966–1.053) but imprecise (SD of O/E = 0.279–0.304) in predicting the fauna actually observed in validation sites. All models were subtly more precise than a null model (mean 1.038, SD 0.343) or the typology (1.046, 0.327). The taxon‐specific NPMR model was slightly more precise than the other three models based on site groupings. 4. The O/E values correlated relatively weakly (r = 0.55–0.86) among the approaches, which thus produced contrasting lake‐specific assessments, despite their seemingly comparable performances. Indeed, typology, suggesting that MI assemblages were impaired in 56% of test sites, was more sensitive than the other approaches (26–46%) as an indicator of human‐induced deterioration. However, this greater ostensible sensitivity seemed to be biased, as lake morphometry, a main driver of natural community variation, remained uncontrolled by the typology. 5. Generally, our exercise illustrates the inconclusiveness of the common validation criteria for the assessment methods. The apparent poor predictability of the profundal fauna, irrespective of the method, may partly stem from large observation error, which could be alleviated by more intensive sampling. However, instead of an O/E‐taxa index, some other metric encompassing quantitative aspects might be preferable for assessing these species‐poor communities.     

Effects of locally rare taxa on the precision and sensitivity of RIVPACS bioassessment of freshwaters

1. An overall aim in freshwater bioassessment is to use biological methods, metrics and forms of indices which are precise, in that they give repeatable results between replicate samples, but which are also sensitive to changes in environmental impacts and stresses. Here we studied the effects of excluding taxa with site‐specific River Invertebrate Prediction and Classification System (RIVPACS)‐type model expected probabilities less than (or equal to) a threshold P_t (0.0, 0.1, 0.2,…,0.9) on the value, precision and power to detect biological effects of environmental stress using the observed to expected ratios (O/E) of biotic indices used to assess the ecological status of U.K. river sites. 2. Amongst the 614 high quality GB RIVPACS reference sites, excluding taxa with low expected probabilities of occurrence gave less total variation (i.e. lower SD) in the estimates O/E for number of taxa (O/E_TAXA) and the average score per taxon (O/E_ASPT). 3. A separate analysis of a replicated sampling study of sites from a wide range of physical types and qualities revealed that sampling variances in O/E for reference condition sites decreased as more locally rare taxa were excluded (but only up to P_t = 0.5 for O/E_ASPT). However, for moderately impacted and poor quality sites, estimates of both O/E_TAXA and O/E_ASPT based on all (P_t = 0.0) or most taxa (i.e. P_t ≤ 0.3) had lower sampling variances and were more precise. 4. Within a very large independent set of test sites with a wide range of perceived levels of environmental stress, increasing the threshold P_t led to systematic compression of the realised O/E scale towards unity. Specifically, with increasing threshold, O/E values >1 are on average reduced, while O/E values <1 have a tendency to be higher and closer to unity (with the exception of O/E_ASPT for the most severely stressed sites). 5. Accuracy and statistical power to detect environmental stress (measured by the percentage of stressed sites with O/E below the lower 10‐percentile value for reference sites) was very similar using O/E_TAXA for P_t up to 0.7. Using O/E_ASPT, power to detect overall general stress decreased slowly as P_t was increased; the rate of fall in power was slightly faster when restricted to sites subject to moderate or severe stress from organic inputs. 6. Taxa which are more sensitive to (organic) stresses [i.e. have high Biological Monitoring Working Party (BMWP) scores] tend to be naturally less widespread (i.e. amongst reference sites) and thus were found to have considerably lower average site‐specific expected probabilities; this may explain why the use of higher thresholds P_t can exclude more such sensitive taxa and lead to underestimation of the extent of impacts. 7. The standard U.K. RIVPACS sampling and sample processing procedures aim to identify all taxa within a sample. This may lead to a longer distribution tail of rarer (low probability) taxa than sampling methods based on a fixed count subsample and influence the practical effects of excluding rare taxa with low expected probabilities from bioassessments.     

Seasonal and geographic climate variabilities during the Last Glacial Maximum in North America: Applying isotopic analysis and macrophysical climate models

Climate models provide estimates of climatic change over periods of time in the ancient past. Macrophysical climate models (MCM) differ from the more widely used general circulation models (GCM), in that MCMs provide temporally high-resolution (~ 100 years) and site-specific estimations of monthly values of climate variables such as temperature and precipitation. In this paper, seasonal changes in climate variables are modeled for six ¹⁴C-dated fossil localities in North America. Five of these localities represent the time of maximum extent of ice during the most recent glacial episode, the Full Glacial (25 + –15 ka), including one at the peak of the Last Glacial Maximum (17–15 ka). The other locality represents the time as the ice began to recede, the Late Glacial (15–11 ka). Seasonal variations in temperature and precipitation modeled by MCM are herein compared with interpretations of seasonal variation based upon oxygen isotopes from serially sampled hypsodont teeth (mostly Equus and Bison) collected from each of these localities. Additionally, the MCM-modeled seasonal variations are used to predict the expected abundances of different plant functional groups (PFG) during those times, especially C3 and C4 functional groups, using modern relationships. These predictions are compared with carbon isotopic values from the same teeth. The importance of atmospheric pCO² for the relative abundance of plants utilizing the C4 metabolic pathway is discussed, given that glacial episodes are known to have been times of lower atmospheric pCO². Interpretations of seasonal variability and the relative abundance of C3 versus C4 vegetation based upon isotopes from tooth enamel are in broad agreement with predictions using the MCM and the modern distribution of PFGs with climate variables. The influence of pCO² on the distribution of C4 vegetation during glacial times appears to be negligible.     

Independent, Interactive, and Species-Specific Responses of Leaf Litter Decomposition to Elevated CO2 and O3 in a Northern Hardwood Forest

The future capacity of forest ecosystems to sequester atmospheric carbon is likely to be influenced by CO₂-mediated shifts in nutrient cycling through changes in litter chemistry, and by interactions with pollutants like O₃. We evaluated the independent and interactive effects of elevated CO₂ (560 μl l⁻¹) and O₃ (55 nl l l⁻¹) on leaf litter decomposition in trembling aspen (Populus tremuloides) and paper birch (Betula papyrifera) at the Aspen free air CO₂ enrichment (FACE) site (Wisconsin, USA). Fumigation treatments consisted of replicated ambient, +CO₂, +O₃, and +CO₂ + O₃ FACE rings. We followed mass loss and litter chemistry over 23 months, using reciprocally transplanted litterbags to separate substrate quality from environment effects. Aspen decayed more slowly than birch across all treatment conditions, and changes in decomposition dynamics of both species were driven by shifts in substrate quality rather than by fumigation environment. Aspen litter produced under elevated CO₂ decayed more slowly than litter produced under ambient CO₂, and this effect was exacerbated by elevated O₃. Similarly, birch litter produced under elevated CO₂ also decayed more slowly than litter produced under ambient CO₂. In contrast to results for aspen, however, elevated O₃ accelerated birch decay under ambient CO₂, but decelerated decay under enriched CO₂. Changes in decomposition rates (k-values) were due to CO₂- and O₃-mediated shifts in litter quality, particularly levels of carbohydrates, nitrogen, and tannins. These results suggest that in early-successional forests of the future, elevated concentrations of CO₂ will likely reduce leaf litter decomposition, although the magnitude of effect will vary among species and in response to interactions with tropospheric O₃.     

Ecosystem respiration depends strongly on photosynthesis in a temperate heath

We measured net ecosystem CO₂ flux (F _n) and ecosystem respiration (R _E), and estimated gross ecosystem photosynthesis (P _g) by difference, for two years in a temperate heath ecosystem using a chamber method. The exchange rates of carbon were high and of similar magnitude as for productive forest ecosystems with a net ecosystem carbon gain during the second year of 293 ± 11 g C m⁻² year⁻¹ showing that the carbon sink strength of heather-dominated ecosystems may be considerable when C. vulgaris is in the building phase of its life cycle. The estimated gross ecosystem photosynthesis and ecosystem respiration from October to March was 22% and 30% of annual flux, respectively, suggesting that both cold-season carbon gain and loss were important in the annual carbon cycle of the ecosystem. Model fit of R _E of a classic, first-order exponential equation related to temperature (second year; R ² = 0.65) was improved when the P _g rate was incorporated into the model (second year; R ² = 0.79), suggesting that daytime R _E increased with increasing photosynthesis. Furthermore, the temperature sensitivity of R _E decreased from apparent Q ₁₀ values of 3.3 to 3.9 by the classic equation to a more realistic Q ₁₀ of 2.5 by the modified model. The model introduces R _photo, which describes the part of respiration being tightly coupled to the photosynthetic rate. It makes up 5% of the assimilated carbon dioxide flux at 0°C and 35% at 20°C implying a high sensitivity of respiration to photosynthesis during summer. The simple model provides an easily applied, non-intrusive tool for investigating seasonal trends in the relationship between ecosystem carbon sequestration and respiration.     

Genetic structure of refugial populations of the temperate plant Shortia rotundifolia (Diapensiaceae) on a subtropical island

Continental island systems harbour relict biota and populations that might have migrated during glacial periods due to the formation of landbridges. Here we analysed the genetic structure of relict populations of the temperate plant Shortia rotundifolia on the subtropical island of Iriomotejima, Japan. This plant, which inhabits riparian environments, is designated “near threatened”. Only five extant populations have been found on the island. Our analyses of 10 nuclear microsatellite loci detected genetic diversity of H _E = 0.488 and H _O = 0.358 for all populations of S. rotundifolia on the island. A high inbreeding coefficient for all populations together (F _IS = 0.316) and each population separately (F _IS = 0.258–0.497) might be attributable to crossing among closely related descendants within a population, an idea that is supported by the relatedness coefficient. These results and an examination of the populations’ demographic histories suggest that the extant populations on Iriomotejima have not experienced a recent population bottleneck. The five extant populations were genetically differentiated (F _ST = 0.283; P < 0.001), suggesting low seed dispersal by gravity and/or low pollen flow via pollinators in the riparian environment. In addition, population differentiation was not related to genetic distance, implying that at one time, ancestral populations might have been distributed over a wider area of the island. However, population fragmentation and range contraction might have occurred at random during the postglacial period.     

Phenology of first and peak emergence of Aphthona lacertosa and A. nigriscutis: Two flea beetles introduced for biological control of leafy spurge, Euphorbia esula L.

Nonlinear models were used to estimate first emergence and peak abundance dates for Aphthona lacertosa Rosenhauer and A. nigriscutis Foudras, two flea beetles introduced to control leafy spurge, Euphorbia esula L., in North America. For model development, 26 field sites were sampled for flea beetle abundance at weekly intervals for eight weeks in three western Minnesota counties in 2000, 2001, and 2002. A three-parameter Weibull function, fit to observed cumulative probability distributions, were used to predict accumulated degree-days (ADD) to first emergence. Bias testing indicated the Weibull function provided a useful estimate of first emergence for A. lacertosa (304 ADD, lower developmental threshold 7.5 °C), but failed to produce a useful estimate for A. nigriscutis. A third-order polynomial was used to approximate seasonal abundance and predict peak abundance for each species. Estimated ADD to peak abundance of A. lacertosa was 594 ± 24 (DD > 7.5 °C) and 670 ± 15 (DD > 9.3 °C) for A. nigriscutis. Models were validated with additional data sets from Minnesota, Montana, and North Dakota. Estimated date of peak emergence provided useful predictions of peak emergence for Minnesota and North Dakota, but failed to predict peak emergence in Montana. We speculate that variation in climate and environmental conditions between Midwestern states and Montana were responsible for differing emergence patterns. We conclude that phenology models should be developed regionally to provide useful predictions of peak emergence for land managers. Maps were developed for Minnesota to spatially display predicted dates of peak abundance for A. lacertosa and A. nigriscutis.     

Plant-available P for Maize and Cowpea in P-deficient Soils from the Nigerian Northern Guinea Savanna – Comparison of E- and L-values

There are several indications that legumes are capable of accessing sparingly soluble phosphorus (P) in the soil through root-induced processes. We hypothesize that this plant-induced mobilization of P can be demonstrated if the plant accessible P assessed by isotopic dilution (‘L-value’) exceeds the corresponding values assessed in soil extracts (‘E-values’). A greenhouse experiment was set up to assess if L/E ratios are affected by P supply and by crop type. The L- and E-values were determined in three P-deficient soils of the Nigerian Northern Guinea savanna (NGS), applied with various rates of TSP, for two cowpea varieties (Vigna unguiculata L., cv Dan-Ila and cv IT-82D-716) and maize (Zea mays L., cv oba super I) as a reference. Plants grown in control soils were severely P-deficient. Plant growth and shoot P uptake significantly increased with increasing P application in all three soils and for all crops, but relative yield and shoot-P responses to P application were similar between maize and cowpea. Both L- and E-values increased with increasing P application. Average L/E ratios for maize were 1.4±0.3 and were unaffected by the P application. For cowpea in contrast, L/E ratios were 3.1±0.2 (significantly larger than one) in one of the three control soils and significantly decreased to 1.3±0.1 at largest P supply. Elevated L/E ratios in cowpea were not associated with an increase in P uptake compared to the other two control soils in which no increase in L/E ratio was observed. It is concluded that cowpea is able to access non-labile P under P-deficient conditions. However, this process cannot overcome P deficiency in these soils, probably because P uptake is limited by the small P concentration in the soil solution (1–2 μg P L⁻¹) and this limitation is not overcome by an increase in the accessible soil P quantity (L-value).     

Tandem repeat markers for population genetic studies of the protected California tiger salamander (<Emphasis Type="Italic">Ambystoma californiense</Emphasis>)

Habitat loss is the single greatest threat to persistence of the critically threatened California tiger salamander (Ambystoma californiense). To aid management plans that designate critical habitat for this species, I developed and characterized 21 tetranucleotide microsatellite markers using two native populations in Santa Barbara and Alameda Counties. Allelic variation and average heterozygosities were lower in the endangered Santa Barbara population (allele range 1–4, mean 2.4; H _O = 0.308 H _E = 0.288) compared with the threatened Alameda population (allele range 2–10, mean 6.7; H _O = 0.712, H _E = 0.722). In-depth population studies using these markers will provide vital information for plans to assign critical habitat that optimize gene flow among breeding populations, as well as for identifying non-native hybrid genotypes that threaten native A. californiense stocks. Beyond the conservation goals for A. californiense, the close phylogenetic relationships within the tiger salamander complex also suggest a broad utility for population studies using these markers.     

Biocontrol of tomato wilt by Penicillium oxalicum formulations in different crop conditions

Eight formulations of Penicillium oxalicum (FOR1 to FOR8) were obtained by the addition of various ingredients, in two separate steps of the production and drying of P. oxalicum conidia. These formulations were then evaluated against tomato wilt in three glasshouse (G1 to G3) and two field (F1 and F2) experiments. All formulations were applied to seedlings in seedbeds 7 days before transplanting at a rate of 10⁷ spores g⁻¹ seedbed substrate. The conidial viability of each formulation was estimated by measuring germination just after fluid bed-drying, before seedbed application and after 1 and 2 years of storage at 4 °C under vacuum. The densities of P. oxalicum were estimated in the seedbed substrate and in the rhizosphere of three plants per treatment just before transplanting. Initial conidial viability of formulations just after fluid bed-drying was approx. 80%, except for FOR1, FOR4, and FOR7 which were 60%. The initial viability was maintained up to 40–50% for 2 years of storage at 4 °C under vacuum, except for FOR1. All formulations had 50% viability at application time. Populations of P. oxalicum in the seedbed substrate just before transplanting were >10⁶ cfu g⁻¹ soil in G3 and F2; populations in rhizosphere were also >10⁶ cfu g⁻¹ fresh root, except for FOR3, FOR5, and FOR6 in G2. A range of 22–64% of disease reduction was observed with all formulations, although these reductions were not significant (p = 0.05) for FOR1, FOR4, and FOR5 in any experiment. Contrast analysis showed significant differences between biological treatments and untreated control (p = 0.05) in all experiments, but no significant differences between biological and chemical treatments. Initial conidial viability of P. oxalicum in formulations and populations of P. oxalicum in the seedbed substrate explained 78.26% of the variability in P. oxalicum populations in tomato rhizosphere before transplanting. Disease incidence in untreated plants was negatively correlated (r = −0.54) with the percentage of disease control. The relationship between the viability of formulations, the populations of P. oxalicum in seedbed and rhizosphere, and the control of tomato wilt is discussed.     

Low levels of quantitative and molecular genetic differentiation among natural populations of Medicago ciliaris Kroch. (Fabaceae) of different Tunisian eco-geographical origin

In this paper, we analyze the genetic variability in four Tunisian natural populations of Medicago ciliaris using 19 quantitative traits and six polymorphic microsatellite loci. We investigated the amplification transferability of 30 microsatellites developed in the model legume M. truncatula to M. ciliaris. Results revealed that about 56.66% of analyzed markers are valuable genetic markers for M. ciliaris. The most genetic diversity at quantitative traits and microsatellite loci was found to occur within populations (>80%). Low differentiations among populations at quantitative traits Q _ST  = 0.146 and molecular markers F _ST  = 0.18 were found. The majority of measured traits exhibited no significant difference in the level of Q _ST and F _ST . Furthermore, significant correlations established between these traits and eco-geographical factors suggested that natural selection should be invoked to explain the level of phenotypic divergence among populations rather than drift. There was no significant correlation between population differentiation at quantitative traits and molecular markers. Significant spatial genetic structure consistent with models of isolation by distance was detected within all studied populations. The site-of-origin environmental factors explain about 9.07% of total phenotypic genetic variation among populations. The eco-geographical factors that influence more the variation of measured traits among populations are the soil texture and altitude. Nevertheless, there were no consistent pattern of associations between gene diversity (He) and environmental factors.     

Development of 15 polymorphic microsatellite loci from <Emphasis Type="Italic">Garrulax morrisonianus</Emphasis> (Timaliidae), an endemic avian species of Taiwan

Garrulax morrisonianus, an endemic avian species of Taiwan, inhabits evergreen forests at high elevations of 2,000–3,952 m. In this study, we developed 15 microsatellite primer pairs for genetic study. These markers were screened for 52 samples collected from wild populations of different geographical regions. The number of alleles ranged from 4 to 13. The expected (H _E) and observed (H _O) heterozygosities were 0.430–0.725 and 0.000–0.500, respectively. All loci were significantly deviated from Hardy-Weinberg equilibrium due to the heterozygote deficiency. The authors Tzen-Yuh Chiang, Sao-Cheng Lao, Ya-Fu Lee contributed to the study equally.     

Eight SSR loci from Oyster Crassostrea gigas EST database and cross-species amplification in C. plicatula

Oyster (Crassostrea plicatula) is widely distributed in coastal areas of China. We developed and evaluated simple sequence repeat (SSR) markers from expressed sequence tags (ESTs) of Crassostrea gigas and to amplify EST-SSR in C. plicatula. Characteristics of eight EST-SSR loci were investigated using 37 wild-type C. plicatula individuals. The number of alleles per locus ranged from four to six. The observed heterozygosity (H _O) ranged from 0.1892 to 0.7027 (0.3919 on average) and the expected heterozygosity (H _E) ranged from 0.6068 to 0.7656 (0.7039 on average) (P < 0.05). The average observed heterozygosity was much lower than the average expected heterozygosity. All loci except C15 departed from Hardy-Weinberg equilibrium (P < 0.05) significantly. Contribution of the eight EST-SSR primers presented here will provide necessary and powerful molecular tools for management and conservation studies on the species of oysters in the future.     

Comparative analysis of <Emphasis Type="Italic">Pasteurella pneumotropica</Emphasis> isolates from laboratory mice and rats

Selected biochemical and genetic characteristics of the wild-type strains of Pasteurella pneumotropica isolated from mice and rats were investigated and compared in order to determine the significant differences among the isolates. The isolates were divided into six groups on the basis of the patterns of carbon source utilization in the host rodents. The genome sizes were determined by electrophoretic analysis, and the mean genome size of the isolates from mice was larger than that of the isolates from rats (P < 0.05). Cluster analysis of the rpoB sequences discriminated five clusters; the differences might have correlated with the host associations. Principal component analysis (PCA) based on both the biochemical and genetic characteristics revealed total 44 strains discriminated into three groups comprising the host-dependent and host-independent groups. Although the P. pneumotropica isolates were mainly classified on the basis of the host rodents by the examinations, the existence of isolates that could not be discriminated on the basis of the host rodents alone was confirmed by the PCA. These results indicated that the P. pneumotropica isolates could be further classified by taxonomic analysis and also suggested the existence of a host-independent group in addition to the host-dependent groups.     

Using molecular genetics to identify immature specimens of the weevil Ceratapion basicorne (Coleoptera: Apionidae)

Molecular analyses can play a primary role in the process of host specificity evaluation at species and population levels. Here we present an example of their application with a promising candidate biological control agent for yellow starthistle, Centaurea solstitialis L. Although it is highly host specific, Ceratapion basicorne (Coleoptera: Apionidae) can develop on safflower in laboratory tests. A field experiment was conducted to further evaluate host plant specificity; however, it was not possible to rear all larvae to the adult stage, which was necessary for species determination. Therefore molecular genetic methods were used to identify immature specimens. A 731 bp fragment of mtDNA cytochrome C oxidase I gene (COI) was sequenced from 41 individuals of C. basicorne and four congeners: Ceratapion orientale, Ceratapion onopordi, Ceratapion penetrans and Ceratapion scalptum. Intraspecific variability ranged from 0.0% to 0.2%, and interspecific divergences ranged from 1.7% to 17.6%. All larvae that were sequenced from the field study, clearly matched one of the five species, enabling us to unambiguously identify them. Use of molecular genetics to identify larvae should also help the process of foreign exploration, enabling the identification of field-collected larvae, which often provide more reliable host plant associations than field collected adults.     

Predictive Models for Freshwater Biological Assessment: Statistical Approaches,Biological Elements and the Iberian Peninsula Experience: A Review

In the last three decades, predictive models have been developed and applied worldwide for freshwater bioassessment. They consist of statistical tools that follow the concept of the Reference Condition Approach. Composed of several sequential steps, these assessment tools assess the deviation of given site assemblages from the expected biological condition in the absence of human disturbance. The most common approaches (RIVPACS/AUSRIVAS and BEAST) are based on a posteriori classifications that use the biological composition of a community to classify reference sites in groups, and afterwards to establish which environmental features best discriminate the biological groups obtained. Here, we review the predictive modeling procedures used in freshwaters bioassessment (RIVPACS/AUSRIVAS, BEAST, ANNA, Artificial Neural Networks, Bayesian Belief Networks and others) as well as the biological elements to which they have been applied. We also review the Spanish and Portuguese experiences in the development and application of predictive models, with particular attention to regional environmental conditions, the different modeling approaches, and the available implementation tools. Moreover, and considering the natural continuity within the Iberian Peninsula (which include several transnational rivers), we discuss the possibilities of the development of common predictive models across the region, considering all factors that may influence their performance, such as the target scale used to develop the models (regional or peninsular); common reference criteria; sampling and sorting procedures; the taxonomic resolution used in the models; the temporal variability (mainly in the Iberian Mediterranean region); and the biological elements to consider. We concluded that there are good technical conditions for the implementations of a common predictive approach throughout the Iberian Peninsula, which should allow a global biological assessment of streams with different biological elements and seasons that could be used by water managers in the context of the Water Framework Directive. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

DIATMOD: diatom predictive model for quality assessment of Portuguese running waters

A predictive model for diatoms based on an adaptation of the River Invertebrate Prediction and Classification System/Australian River Assessment System approaches was evaluated as an effective tool for measuring stream ecological quality. This type of model was originally developed in UK and later in Australia and is extensively used to obtain ecological quality assessments with macroinvertebrates. The first step for the model construction was the definition of six consistent reference biological groups (ANOSIM: Global R = 0.77; P < 0.001) after classification (UPGMA) and ordination (nMDS) of 120 reference sites containing 254 different diatom taxa (species and infra-specific rank). A set of five environmental variables (slope, hydrological regime, mean annual temperature, mean annual precipitation and alkalinity) correctly discriminated 67% of reference sites (stepwise forward discriminant analysis, Jackknifed classification). The model was statistically accurate (slope = 1.07, intercept = −0.68, R ² = 0.65) and was validated by an independent set of reference data (13 reference sites; 70% correct answers). In addition, the model was tested by running data from 113 potentially disturbed sites. The model (DIATMOD) was well correlated with a general abiotic degradation gradient (Spearman correlations, R ² = 0.53, P < 0.001; and PCA analysis) and also with several specific pressure variables such as nitrates, phosphates, urban area, connectivity and land use (P < 0.001). Most diatom indices assess chemical contamination and we showed here that through predictive modelling the potential of diatoms as bioindicators increases as they also responded to hydromorphological changes. Further investigation on model potential consists in: testing different probability levels for taxa inclusion (here it was >0.5 as the most common models); comparing with alternative classification systems; assessing the influence of substrate type and seasonal variation in assessments.     

Multiple host use by a sap-sucking membracid: population consequences of nymphal development on primary and secondary host plant species

Aconophora compressa is a gregarious, sap-sucking insect that uses multiple host plant species. Nymphal host plant species (and variety) significantly affected nymphal survival, nymphal development rate and the subsequent size and fecundity of adults, with fiddlewood (Citharexylum spinosum) being significantly best in all respects. Nymphs that developed on a relatively poor host (Duranta erecta var “geisha girl”) and which were moved to fiddlewood as adults laid significantly fewer eggs (mean ± SE = 836 ± 130) than those that developed solely on fiddlewood (1,329 ± 105). Adults on geisha girl, regardless of having been reared as nymphs on fiddlewood or geisha girl, laid significantly fewer eggs (342 ± 83 and 317 ± 74, respectively) than adults on fiddlewood. A simple model that incorporates host plant related survival, development rate and fecundity suggests that the population dynamics of A. compressa are governed mainly by fiddlewood, the primary host. The results have general implications for understanding the population dynamics of herbivores that use multiple host plant species, and also for the way in which weed biological control host testing methods should be conducted. Handling Editor: Robert Glinwood