Ciliate communities in shallow groundwater: seasonal and spatial characteristics

1. Ciliated protozoans (Phylum Ciliophora) were collected from five sites in a shallow groundwater system in southern Ontario, Canada over a 13‐month period: one at the spring source, two along the channel banks, and two in the stream channel. Ciliates and environmental data were collected from surface water and at five depths into the sediment, located at 20, 40, 60, 80 and 100 cm. 2. Species richness was high (170 ciliate species belonging to 89 genera were identified) and variable, both spatially and temporally. Highest species richness (86) occurred between 20 and 60 cm, and typically decreased below 60 cm. 3. Ciliate densities were also seasonally and spatially variable. Densities peaked in March between 40 cm (as high as 69 900 cells L⁻¹) and 60 cm, and again in May and June at 80 and 100 cm. Densities were lowest in winter. The surface‐water ciliate community had a different species composition and lower population densities. 4. At all depths, small (<50 μm) bacterivorous ciliates typically dominated, but omnivorous and predatory species were also present (combined, up to 30% of the average density). 5. Several ciliate genera, traditionally considered planktonic, occurred at low densities from 40 cm down to 100 cm. 6. Ordination analysis indicated that the main factors influencing the shallow groundwater ciliate communities were depth and temperature. 7. Dissolved oxygen also appeared to influence these communities in that they typically comprised genera that preferred either low‐oxygen or anaerobic conditions.     

Drivers of host plant shifts in the leaf beetle Chrysomela lapponica: natural enemies or competition?

1. The leaf beetle, Chrysomela lapponica, originally uses the salicyl glucosides (SGs) of its host plants to sequester salicylaldehyde, which serves as a defence against generalist enemies but attracts specialist enemies. However, some populations of C. lapponica have shifted to SG‐poor hosts, and their secretions do not contain salicylaldehyde. 2. In was suggested that beetles shift to SG‐poor hosts to escape from specialist enemies. To test this hypothesis, we compared field mortality between two populations of C. lapponica that were associated with SG‐rich willow, Salix myrsinifolia (Kola Peninsula and Finland) and two populations that fed on SG‐poor willows, S. glauca (Ural) and S. caprea (Belarus). 3. Mortality from generalist enemies was significantly higher in Belarus than in three other populations, whereas mortality from specialists did not differ among populations. A specialist predator (syrphid fly larvae, Parasyrphus nigritarsis) and specialist parasitoids (phorid flies, Megaselia spp.) were attracted to the secretions of larvae reared on both SG‐rich and SG‐poor hosts. 4. Feeding on leaves of S. caprea and S. myrsinifolia both previously damaged by leaf puncturing and by the larvae of potentially competing species Chrysomela vigintipunctata, decreased the weight and prolonged the development of C. lapponica. 5. Thus, populations of C. lapponica that have shifted to SG‐poor willow species did not obtain enemy‐free space because specialist enemies have developed adaptations to herbivores that switched to a novel host plant. We suggest that in some populations host plant shift was favoured by interspecific competition with the early season SG‐using specialist, C. vigintipunctata.     

Climate change-predicted shifts in the temperature regime of shallow groundwater produce rapid responses in ciliate communities

The ciliates living in a shallow groundwater system in southern Ontario, Canada were subjected to an in situ temperature manipulation over 14 months. Ciliates were collected from the bed surface of a small springbrook and from interstitial water collected at five depths beneath its surface. Mean temperature elevations established at each depth (?20, ?40, ?60, ?80, and ?100 cm) between the experiment's control and treatment blocks were 1.9, 3.5, 3.9, 3.8, and 3.6 °C, respectively, and were based on global warming projections for the region. In total, 160 species of ciliate belonging to 85 genera were identified. Overall, the treatment block had a higher density (6510±342 cells L^?1; ±1 SE) than the control (5797±237 cells L^?1), but densities were both vertically and longitudinally variable. Control densities decreased with depth, whereas treatment densities were more equal among depths. Total species richness showed no significant difference between blocks when combining all sampling dates and depths, although species composition changed. The ciliate community was dominated by small (15–50 μm), followed by medium (50–200 μm), and only a few large‐sized (>200 μm) species. Small ciliates contributed 82–97% of the total density. Small ciliates also contributed more to the treatment (94%) than the control block (88%). The most common ciliate feeding groups were bacterivores, omnivores, predators, and algae‐diatom feeders, with bacterivores being most dominant (83–99% of the total numbers collected). Ordination analyses revealed that ciliate distribution was strongly correlated with groundwater temperature, although dissolved oxygen level, concentrations of ammonia and nitrate, and depth also appeared to be influential. Peak densities of many species occurred in either the control or treatment blocks, but not in both. The benefits of using ciliates as a proxy for higher, much longer‐lived, eukaryotes in climate change studies are discussed.     

Identification of mutants in phosphorus metabolism

Phosphorus availability is often limiting for plant growth. However, little is known of the pathways and mechanisms that regulate phosphorus (P) uptake and distribution in plants. We have developed a screen based on the induction of secreted root acid phosphatase activity by low‐P stress to identify mutants of Arabidopsis thaliana with defects in P metabolism. Acid phosphatase activity was detected visually in the roots of A. thaliana seedlings grown in vitro on low‐P medium, using the chromogenic substrate, 5‐bromo‐4‐chloro‐3‐indolyl‐phosphate (BCIP). In low‐P stress conditions the roots of wild‐type plants stained blue, as the induced root acid phosphatase cleaved BCIP to release the coloured product. Potential mutants were identified as having white, or pale blue, roots under these conditions. Out of approximately 79 000 T‐DNA mutagenised seedlings screened, two mutants with reduced acid phosphatase staining were further characterised. Both exhibited reduced growth and differences in their P contents when compared to wild‐type A. thaliana. The mutant with the most severe phenotype, pho3, accumulated high levels of anthocyanins and starch in a distinctive visual pattern within the leaves. The phenotypes of these mutants are distinct from two previously identified phosphorus mutants (phol and pho2) and from an acid phosphatase deficient mutant (pupl) of A. thaliana. This suggested that the screening method was robust and might lead to the identification of further mutants with the potential for increasing our understanding of P nutrition.     

Application of PCR for Specific Diagnosis of Leptospirosis in Humans in Ukraine

Leptospirosis remains one of the most widespread zoonotic diseases in the world and Ukraine, in particular. Ukrainian clinicians have been faced with early detection of the disease due to the availability of only a serological method for routine diagnostics in Ukraine, namely the microscopic agglutination test (MAT). This paper demonstrates the first results of the complex application of MAT and polymerase chain reaction (PCR) for routine verification of leptospirosis, which were first applied simultaneously in Lviv Oblast of Ukraine in 2016. We examined the sera of 150 patients clinically suspected of leptospirosis, 31 of whom were treated at the Lviv Oblast Clinical Hospital for Infectious Diseases (LOCHID). The application of PCR during the first seven days of the disease allowed increasing the share of confirmed leptospirosis cases by 16,1% in patients that were treated in LOCHID during 2016–2017.Key words: Leptospirosis, diagnostics, microagglutination test, polymerase chain reaction     

Proton cellular influx as a probable mechanism of variation potential influence on photosynthesis in pea

Electrical signals (action potential and variation potential, VP) caused by environmental stimuli are known to induce various physiological responses in plants, including changes in photosynthesis; however, their functional mechanisms remain unclear. In this study, the influence of VP on photosynthesis in pea (Pisum sativum L.) was investigated and the proton participation in this process analysed. VP, induced by local heating, inactivated photosynthesis and activated respiration, with the initiation of the photosynthetic response connected with inactivation of the photosynthetic dark stage; however, direct VP influence on the light stage was also probable. VP generation was accompanied with pH increases in apoplasts (0.17–0.30 pH unit) and decreases in cytoplasm (0.18–0.60 pH unit), which probably reflected H⁺‐ATPase inactivation and H⁺ influx during this electrical event. Imitation of H⁺ influx using the protonophore carbonyl cyanide m‐chlorophenylhydrazone (CCCP) induced a photosynthetic response that was similar with a VP‐induced response. Experiments on chloroplast suspensions showed that decreased external pH also induced an analogous response and that its magnitude depended on the magnitude of pH change. Thus, the present results showed that proton cellular influx was the probable mechanism of VP's influence on photosynthesis in pea. Potential means of action for this influence are discussed.