Evaluation of an internal positive control for Cryptosporidium and Giardia testing in water samples

AIMS: An internal positive control for Cryptosporidium and Giardia monitoring was evaluated for use in routine water monitoring quality control. The control, known as ColorSeed C&G (BTF Pty Ltd, Sydney, Australia), is a suspension containing exactly 100 Cryptosporidium oocysts and 100 Giardia cysts that have been modified by attachment of Texas Red to the cell wall, allowing them to be differentiated from unmodified oocysts and cysts. The control enables recovery efficiencies to be determined for every water sample analysed. METHODS AND RESULTS: A total of 494 water samples were seeded with ColorSeed C&G and with unlabelled Cryptosporidium and Giardia and then analysed. Additionally, the robustness of the ColorSeed labelling was challenged with various chemical treatments. Recoveries were significantly lower for the ColorSeed Texas Red labelled Cryptosporidium and Giardia than recoveries of unlabelled Cryptosporidium and Giardia. However, the differences in recoveries were small. On average ColorSeed Cryptosporidium recoveries were 3.3% lower than unlabelled Cryptosporidium, and ColorSeed Giardia recoveries were 4% lower than unlabelled Giardia. CONCLUSIONS: ColorSeed C&G is suitable for use as an internal positive control for routine monitoring of both treated and raw water samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The small differences in recoveries are unlikely to limit the usefulness of ColorSeed C&G as an internal positive control. The ColorSeed labelling was found to be robust after different treatments.     

Profiling bacterial survival through a water treatment process and subsequent distribution system

AIMS: To profile fractions of active bacteria and of bacteria culturable with routine heterotrophic plate count (HPC) methods through a typical water treatment process and subsequent distribution system. In doing so, investigate how water treatment affects both bacterial abundance and diversity, and reveal the identities of active bacteria not detected by traditional HPC culture. METHODS AND RESULTS: Profiling active fractions was performed by flow cytometric cell sorting of either membrane-intact (BacLight kit) or enzymatically active (carboxyfluorescein diacetate, CFDA) bacteria, followed by eubacterial 16S rDNA-directed PCR and denaturing gradient gel electrophoresis (DGGE). Water treatment significantly reduced active bacterial numbers detected by the BacLight kit and CFDA assay by 2.89 and 2.81 log respectively. Bacterial diversity was also reduced from > 20 DGGE bands in the active fractions of reservoir water to only two bands in the active fractions of finished water. These two bands represented Stenotrophomonas maltophila, initially culturable by HPC, and a Burkholderia-related species. Both species maintained measurable traits of physiological activity in distribution system bulk water but were undetected by HPC. CONCLUSIONS: Flow cytometric cell sorting with PCR-DGGE, to assess water treatment efficacy, identified active bacteria from a variety of major phylogenetic groups undetected by routine HPC. Following treatment S. maltophila and a Burkholderia-related species retained activity and entered distribution undetected by HPC. SIGNIFICANCE AND IMPACT OF THE STUDY: Methods used here demonstrate how water treatment operators can better monitor water treatment plant efficacy and assess distribution system instability by the detection and identification of active bacteria recalcitrant to routine HPC culture.     

Canopy height variation and environmental heterogeneity in the tropical dry forests of coastal Oaxaca,Mexico

Despite its importance for carbon storage and other ecosystem functions, the variation in vegetation canopy height is not yet well understood. We examined the relationship between this community attribute and environmental heterogeneity in a tropical dry forest of southern Mexico. We sampled vegetation in 15 sites along a 100‐km coastal stretch of Oaxaca State, and measured the heights of all woody plants (excluding lianas). The majority of the ca. 4000 individuals recorded concentrated in the 4–8 m height range. We defined three plant sets to describe overall community canopy height at each site: a set including all plants, a set made up by the tallest plants representing 10 percent of all individuals, and a set comprising the 10 tallest plants. For each site we computed maximum height and the mean and median heights of the three sets. Significant collinearity was observed between the seven resulting height variables, but null distributions constructed through bootstrap revealed their different behaviors as functions of species richness and density of individuals. Through linear modeling and a model selection procedure, we identified 21 models that best described the variation in canopy height variables. These models pointed out to soil (measured as PC1 of a principal component analysis performed on 10 soil variables), water stress, and elevation as the main drivers of canopy height variation in the region. In the event of increasing water stress resulting from global climate change, the studied tropical dry forests could become shorter and thus decrease their carbon storage potential.     

Alleviation of water stress effects in cowpea by Bradyrhizobium spp. inoculation

Experiments were carried out to investigate the effects of different degrees of water stress on cowpea in the presence and absence of Bradyrhizobium spp. inoculation and to evaluate physiological responses to stress. The soil used was Yellow Latosol, pH 6.3 and the crop used was cowpea (Vigna unguiculata (L.) Walp.) cv. ‘IPA 204’. Stress was applied continuously by the control of matric potential (ψ _m ) through a porous cup. The lowered soil ψ _m had a direct effect on the N2 fixation, but the strains Bradyrhizobium introduced by inoculation in the cowpea plants were superior to the indigenous strain demonstrating the importance of inoculation in the stressed plants. At the more negative ψ _m plants inoculated with the strains EI 6 formed associations of greater symbiotic efficiency which helped the cowpea plants to withstand drought stress better than the strain BR 2001 and the uninoculated control. The leghaemoglobin concentration was not inhibited in the drought-stressed plants at ψ _m -70 kPa when inoculated with the strain EI 6, which confered a differential degree of drought resistance in plants. The ψ _w declined in the stressed plants reaching values of -1.0 MPa which was sufficient to cause disturbance in nodulation and biomass production. This revised version was published online in June 2006 with corrections to the Cover Date.     

Characterization of photosynthetic pathway of plant species growing in the eastern Tibetan plateau using stable carbon isotope composition

The photosynthetic pathway of plant species collected at Menyuan, Henan, and Maduo sites, east of Tibetan Plateau, China, during the growing season were studied using stable carbon isotopes in leaves. The 232 samples leaves analyzed belonged to 161 species, 30 families, and 94 genera. The δ¹³C values (from −24.6 to − 29.2 ‰) indicated that all the considered species had a photosynthetic C₃ pathway. The absence of plant species with C₄ photosynthetic pathway might be due to the extremely low air temperature characterizing the Tibetan Plateau. The average δ¹³C value was significantly (p<0.05) different between annuals and perennials at the three considered study sites. Hence the longer-lived species had greater water-use efficiency (WUE) than shorter-lived species, that is, longer-lived species are better adapted to the extreme environmental conditions of the Tibetan Plateau.     

Contributions of water transfer energy to protein‐ligand association and dissociation barriers: Watermap analysis of a series of p38α MAP kinase inhibitors

In our previous work, we proposed that desolvation and resolvation of the binding sites of proteins can serve as the slowest steps during ligand association and dissociation, respectively, and tested this hypothesis on two protein‐ligand systems with known binding kinetics behavior. In the present work, we test this hypothesis on another kinetically‐determined protein‐ligand system—that of p38α and eight Type II BIRB 796 inhibitor analogs. The k_on values among the inhibitor analogs are narrowly distributed (10⁴ ≤ k_on ≤ 10⁵ M^?1 s^?1), suggesting a common rate‐determining step, whereas the k_off values are widely distributed (10^?1 ≤ k_off ≤ 10^?6 s^?1), suggesting a spectrum of rate‐determining steps. We calculated the solvation properties of the DFG‐out protein conformation using an explicit solvent molecular dynamics simulation and thermodynamic analysis method implemented in WaterMap to predict the enthalpic and entropic costs of water transfer to and from bulk solvent incurred upon association and dissociation of each inhibitor. The results suggest that the rate‐determining step for association consists of the transfer of a common set of enthalpically favorable solvating water molecules from the binding site to bulk solvent. The rate‐determining step for inhibitor dissociation consists of the transfer of water from bulk solvent to specific binding site positions that are unfavorably solvated in the apo protein, and evacuated during ligand association. Different sets of unfavorable solvation are evacuated by each ligand, and the observed dissociation barriers are qualitatively consistent with the calculated solvation free energies of those sets.     

Ecological determinants of extrapair fertilizations and egg dumping in Alpine water pipits (Anthus spinoletta)

Behavioral ecology has successfully explained the diversityin social mating systems through differences in environmentalconditions, but diversity in genetic mating systems is poorlyunderstood. The difference is important in situations whereparents care for extrapair young (EPY) originating from extrapairpaternity (EPP), extrapair maternity (EPM), and intraspecificbrood parasitism (IBP). In birds, IBP and EPM are rare, butEPP is widespread and highly variable among species and populations.Explanations for this variability are controversial, mainlybecause detailed ecological information is usually lacking inpaternity studies. Here we present results of the first studyto identify the ecological determinants of extrapair activitiesfor both sexes of the same species, the water pipit (Anthusspinoletta). DNA fingerprints of 1052 young from 258 nests revealedEPP in 5.2% of the young from 12.4% of the nests. EPM and IBP,both involving egg dumping (EDP), each occurred in 0.5% of theyoung from 1.9% of the nests. Nests with and without EPY couldnot be distinguished by traits of the breeders and by reproductivesucccess, but they differed with respect to ecology: nests withEPP young were characterized by asynchronous clutch initiation,nests with EPM and IBP young were characterized by higher overlapwith neighboring territories and closer proximity to communalfeeding sites. We suggest that chance events, resulting fromthe temporal and spatial distribution of broods, offer a betterexplanation for the occurence of extrapair activities than femalesearch for genetic or phenotypic benefits. This possibilityof "accidental" extrapair reproduction as an "ecological epiphenomenon"with low potential for selection should also be considered forspecies other than the water pipit.     

The impact of the River Ntahangwa, the most polluted Burundian affluent of Lake Tanganyika, on the water quality of the lake

Sources of pollution at the north end of Lake Tanganyika are concentrated around Bujumbura, Burundi. The impact of River Ntahangwa on the water quality of this part of Lake Tanganyika was compared with that of an unpolluted river, the Mugere. A shallow bay, not influenced by a river, was used as a reference station. Chemical analyses were carried out at four week intervals during nearly a year. Samples were taken at different distances from the shore and at different depths. River impact was detected up to 100 m from both river mouths. Even at 50 m from the mouth of the polluted River Ntahangwa, no important decrease of dissolved oxygen was detected. Chlorophyll-a maxima were found 100 m from the Ntahangwa mouth, but this was also the case 100 m from shore in the unpolluted bay. Lake water at 60 m depth was saturated with oxygen during the dry season, as it used to be in 1956. The rivers studied descent initially within the lake due to greater sediment load and lower water temperature. Nevertheless, it appears that these rivers mix completely in the lake within 100–300 m from the river mouths. The direct effects from organic pollution of the Ntahangwa on the lake seem to be rather negligible. Appropriate anti-erosion measures should reduce massive sediment discharge into the lake.     

Hydraulic architecture and water relations of Spartium junceum branches affected by a mycoplasm disease

Plant water relations, xylem anatomy and the hydraulic architecture of 1‐year‐old twigs of Spartium junceum, both healthy and affected by a phytoplasm disease, were studied. The disease causes twigs to be either shortened (witches broom disease, WBD) or flat (fasciate disease, FD). WBD twigs show a sevenfold increase in total leaf area, smaller and shorter xylem conduits, a higher stomatal conductance (g_l) and a decline of minimum leaf water potentials ( Ψ _l) below the turgor loss point. FD twigs had nearly twice the leaf area of the healthy controls as well as high g_l values and Ψ _l values below the turgor loss point. Moreover, significant differences between healthy and affected twigs in stem stomatal conductance (g_s) and in the total stem area were recorded. Affected twigs die back under drought stress, which is explained by a pronounced loss of hydraulic conductivity of the infected stems (40 and 60%) in FD and WBD as well as by the unfavourable ratio of weighted conduit radius ( Σ r⁴) to total surface area (A_t), so that the efficiency of the stem in supplying the whole transpiring area with water is strongly reduced.