Performance of pine looper Bupalus piniarius larvae under population build-up conditions

In the first year of an outbreak, Bupalus piniarius larvae, encounter intense crowding. In the later stages of larval development, they are forced to feed on the non-preferred current-year needles of Scots pine or even on alternative hosts. It was hypothesized that larval feeding on a non-preferred resource (current-year needles) will negatively affect B. piniarius performance. It was also hypothesized that larval mutual interference (crowding without competition for food) will have negative additive effects. These hypotheses were tested in laboratory and field experiments. Fourth instar larvae were reared singly and under crowded conditions in cohorts of ten. Larvae in both situations were reared on control branches (containing both mature and current-year shoots) and branches containing only current-year shoots. Crowded larvae were reared also on Norway spruce, an alternative host. Crowding and feeding on a non-preferred resource had opposite effects on B. piniarius larval performance. Crowding in the late larval instars enhanced larval performance while absence of the preferred resource had negative effects. Larval growth rate was higher and development time was shorter for larvae exposed to crowded conditions than for solitary larvae. There was, however, no difference between the groups in final pupal weights or survival. Survival was 25% lower for larvae feeding on non-preferred current-year needles and pupal weights 9% lower, compared with results for larvae feeding on mature needles. Larvae feeding on Norway spruce suffered greatly extended development time, 82% lower survival, and resulted in 60% lighter pupae compared with conspecifics on Scots pine. It was concluded that not only quantity but also quality of the available food resource is critical for B. piniarius development.     

Imaging chlorophyll a fluorescence reveals specific spatial distributions under different stress conditions

Chlorophyll a fluorescence has been adopted as a fast, non-invasive, and cheap method to detect stress effects in plants. The majority of these chl-fluorescence measurements have been carried out with ‘clamping’ fluorometers recording punctual chlorophyll a fluorescence at isolated parts of the leaf. However, this method is inherently limited in providing information on the homogeneity of responses to stresses at the leaf or whole plant level. Therefore the purpose of this study was to measure imaging chlorophyll a fluorescence and to compare the temporal and spatial distribution of this emission under allelochemical (2-3H-benzoxazolinone and 3,4-dihydroxybenzaldehyde), thermal and salt, and heavy metal (cadmium, copper and zinc) treatment in the model plant Arabidopsis thaliana (L.) Heynh. The results suggested different spatial distributions for each condition: the two allelochemicals showed inhibition spots at the edges of the oldest leaves and both did not affect the photosynthetic activity of young leaves; treatment with the three heavy metals revealed highly homogenous effects over the whole plant with a quite uniform decrease of maximum PSII efficiency (also in youngest leaves). On the contrary, temperature (heat and cold) and salt stress showed an initial decrease of fluorescence in the tissues around the vascular bundles that lasted between 2 and 3 h depending on the treatment. These irregularities in chlorophyll fluorescence make it difficult to correlate punctual measures (typical for clamping fluorometers) with the effect on the whole plant, ignoring effects that are evident when imaging is used. Therefore these results show that monitoring chlorophyll a fluorescence by imaging improves the measurement of stress effects on treated plants, suggesting that punctual fluorescence measurements do not always reveal the heterogeneity of the stress-related effects in treated plants.     

Efficiency evaluation of two competing foraging modes under different conditions

Various foraging modes are employed by predators in nature, ranging from ambush to active predation. Although the foraging mode may be limited by physiological constraints, other factors, such as prey behavior and distribution, may come into play. Using a simulation model, we tested to what extent the relative success of an ambush and an active predator changes as a function of the relative velocity and movement directionality of prey and active predator. In accordance with previous studies, we found that when both active predator and prey use nondirectional movement, the active mode is advantageous. However, as movement becomes more directional, this advantage diminishes gradually to 0. Previous theoretical studies assumed that animal movement is nondirectional; however, recent field observations show that in fact animal movement usually has some component of directionality. We therefore suggest that our simulation is a better predictor of encounter rates than previous studies. Furthermore, we show that as long as the active predator cannot move faster than its prey, it has little or no advantage over the ambush predator. However, as the active predator's velocity increases, its advantage increases sharply.     

Sea urchin spermatozoa generate at least two reactive oxygen species; the type of reactive oxygen species changes under different conditions

Reactive oxygen species (ROS) cause oxidative stress and act as signal transduction molecules in many cells. Spermatozoa from several mammals generate ROS, which are involved in male infertility and signaling during capacitation. In the present study, we investigated ROS generation by sea urchin spermatozoa at the initiation of motility, during dilution with seawater, and following egg jelly treatment. In seawater containing an ROS indicator, 5-(and 6-)chloromethyl-2',7'-dichlorodihydrofluorescein diacetate (CM-H(2)DCFDA), fluorescence increased after the addition of spermatozoa. The ROS generation rate was dependent upon the dilution ratio and respiratory rate of the spermatozoa. Spermatozoa in sodium-free seawater did not increase fluorescence, but fluorescence did increase with the addition of NaCl. Sodium chloride also led to the initiation of sperm motility and respiration. Using the indicator MitoSOX Red, ROS generation was detected from spermatozoa exposed to egg jelly dissolved in seawater, but not in normal seawater. Moreover, the respiratory inhibitor antimycin A prevented CM-H(2)DCFDA-detectable ROS and increased MitoSox-detectable ROS at a higher concentration. These findings revealed that the ROS generated were of different species, possibly hydrogen peroxide (H(2)O(2)) and superoxide anion (O(-)(2)), and their detected levels were altered by egg jelly. We concluded that sea urchin spermatozoa generate at least two species of ROS depending on the physiological conditions to which they are exposed. It is possible that the major ROS from sea urchin spermatozoa changes during the course of fertilization.     

Removal of Prymnesium parvum (Haptophyceae) cells under different nutrient conditions by clay

In this study, we have shown that the ichthyotoxic Prymnesium parvum (haptophyte) can be successfully removed by spraying the surface with a phosphatic clay/polyaluminium chloride (PAC) mix (final concentration, 4 g l⁻¹ and 5 ppm, respectively). The alga was grown in non-axenic batch cultures with nitrogen deficient, phosphorus deficient and nutrient sufficient media. Sub-samples of the nutrient sufficient culture were diluted to obtain cell abundances equal to those in the nutrient deficient cultures. Clay/PAC removed up to 100% of the cells in the low cell nutrient sufficient treatment after 72 h, but removal in the other treatments was lower (up to 84%). The nitrogen deficient cells were found to be the most toxic, measured as haemolytic activity of the cells (HE₅₀), just prior to the start of the experiment. However, the toxicity of the cells in all treatments was found to fluctuate during the incubation time with a general increase in toxicity towards the end, suggesting that the cells became stressed during sedimentation and/or when trapped in the clay. But the amount of released toxins was always below the detection limit of the haemolytic assay, and the abundance of free-living bacteria, derived from flow cytometer counts, increased throughout the experiment. This suggests that released toxins were either trapped by the clay particles or effectively degraded by the bacterial community. The study shows that the clay method can be efficient in mitigation of P. parvum blooms, but further studies have to be conducted where optimum clay concentrations are determined to ensure that the efficiency is high against nutrient deficient cells and at high cell abundances, i.e. conditions found during blooms in eutrophic coastal waters, and to determine the fate of the cells and their toxins during and after sedimentation.     

Survival capacity in water of Arcobacter species under different temperature conditions

Aims:  To assess the survival capacity in vitro of arcobacters in water at temperatures applied in the food industry.
Methods and Results:  Four strains of each Arcobacter species were inoculated in potable water and water with 1% organic material and stored at 4, 7, 20, 52, 56 and 60°C. Samples were taken at known time points and the numbers of bacteria were determined on Arcobacter -selective medium. All Arcobacter species remained viable for a temperature-dependent period of time, although Arcobacter butzleri displayed a significant longer survival and heat resistance . No significant intraspecies differences were detected, resulting in no definite identification of origin or strain dependency. The survival period for all species was prolonged in the presence of the organic material only for the low temperatures.
Conclusions:  The present study demonstrates that water can act as a reservoir and as a potential source of Arcobacter contamination to humans and animals.
Significance and Impact of the Study:  This study assessed for the first time the survival of all human-related Arcobacter species in water. Particularly A. butzleri showed to be the most robust species with regard to temperature which is interesting as that species is often found in human clinical specimens.     

Reduction of nitroaromatic compounds by different Pseudomonas species under aerobic conditions

Summary Pseudomonas sp. CBS3 converted various nitro-aromatic compounds under aerobic resting-cell conditions to the corresponding amino compounds. Mononitro-compounds were reduced to anilines. 1-Chloro-2,4-dinitrobenzene was reduced via the two possible chloronitroanilines to 4-chloro-1,3-diaminobenzene. In the case of 2,4,6-trinitrotoluene, two monoaminodinitrotoluenes and one diaminomononitrotoluene were obtained. In addition to the reduction, in most cases the amines were partially acetylated. In experiments under an argon atmosphere conversion of the nitro-compounds was as fast as under aerobic conditions. Cells of Pseudomonas sp. CBS3 cultivated on complex medium showed higher nitro-reducing activity than those cultivated on mineral salts medium with 4-chlorobenzoate as substrate, which is normally used as medium for this strain. Several other Pseudomonas species (ATCC 4359, ATCC 23937, ATCC 15005, ATCC 17933) also showed nitro-reducing activities. In crude cell-free extracts of Pseudomonas sp. CBS3 an enzyme catalysing the reduction of nitro-aromatics was detected. The enzyme was inactivated by dialysis and was reactivated by the addition of NADH or NADPH. NADPH was the more efficient co-substrate.Offprint requests to: R. Müller     

Oviposition in Culicoides impunctatus under laboratory conditions

We present a laboratory-based examination of oviposition preference in the Scottish biting midge, Culicoides impunctatus (Diptera: Ceratopogonidae). A variety of oviposition substrates were screened in no-choice and choice bioassays for efficacy in eliciting an egg-laying response. Both upper-layer photosynthetic Sphagnum spp. moss and Juncus articulatus infusions were identified as producing increased oviposition under no-choice conditions. During choice trials against a control of damp cottonwool, upper-layer Sphagnum spp. moss produced a significantly greater egg-laying response. These conclusions are interpreted in terms of possible cues involved in oviposition site selection and assessed for future use in colonisation of this troublesome species.     

Delayed luminescence of Prorocentrum minimum under controlled conditions

Delayed luminescence (DL), also termed delayed fluorescence or delayed light emission, is the phenomenon of long-lived light emission by plants and cyanobacteria after being illuminated with light and put into darkness. Culture growth of three Prorocentrum minimum strains was studied with DL measurements. DL decay kinetics was measured from 1–60 s after a pulse of white light. The strains used were from the Adriatic Sea (PmK), from Chesapeake Bay, USA (D5), and from the Baltic Sea (BAL), cultured at salinity of 32, 16, and 8 (practical salinity scale), respectively. The strains differed in cell size and chlorophyll a content (PmK > D5 > BAL), as well as in DL parameters. The DL results were compared to standard measurements of culture density and carbon content (calculated from biovolumes). DL decay curves had a specific peak, which changed with culture growth and showed more similarities between the strains PmK and D5. The DL intensity increased with cell density and carbon content in a two-stage process, corresponding to the lag and exponential phases of growth. DL intensity was best correlated with carbon content irrespective of strain and is proposed as an estimate of biomass and for differentiating between lag and exponential phases of growth.     

Copulation behaviour of Lygocoris pabulinus under laboratory conditions

As a prerequisite to the elucidation of the sex pheromone of the green capsid bug Lygocoris pabulinus (L.), we studied the bug's reproductive development and behaviour. When kept under long-day conditions (L18:D6) at 25 °C, both males and females start to copulate 4 days after the final moult. Second matings occur in some females, even on consecutive days. The premating period is 10 min on average and copulation lasts 1 to 2 min. At long range, when males are attracted to traps baited with live females, we did not observe a specific calling position of the females. At short range, a characteristic courtship behaviour of the females was not observed either. Males showed a characteristic vibration of the abdomen, which was repeated several times before copulation. This behaviour can be used as a bioassay to test potential sex pheromone compounds.     

Photosynthetic response of two tropical liana species grown under different irradiances

We investigated the characteristics of gaseous exchanges and chlorophyll a fluorescence under different irradiances in two liana species Canavalia parviflora Benth. (Fabaceae) and Gouania virgata Reissk (Rhamnaceae), both of a semi-deciduous tropical forest of Southeast Brazil. We used cultivated plants growing under irradiances of 100, 40, 10, and 1.5 % of the photosynthetic photon flux density (PPFD). Higher net photosynthetic rates (P _N) were observed during early morning under full sunlight. After this, reduced P _N values were recorded due to pronounced stomatal closure. In Canavalia, the gas exchange responses diminished concomitant with reduced irradiance. Gouania exhibited a narrower range of response, with high P _N values even at 10 % PPFD. Marked reduction of the effective photochemical yield (ΔF/F_m’) near midday was observed, followed by increases in the non-photochemical quenching for both species under full sunlight. Despite the common occurrence of these species in open areas of the forest, both were able to maintain relatively high P _N in shaded environments. We suggest that lianas present an intermediate physiological behaviour between shade and non-shade tolerant species.     

Susceptibility of Cydia pomonella to Bacillus thuringiensis under laboratory and field conditions

The delta-endotoxin of 12 strains in 10 subspecies of Bacillus thuringiensis Berliner was highly active against Cydia pomonella (L.) when assayed under laboratory conditions on artificial diet. These results could not be confirmed in the field.The disappointing results obtained under field conditions are due to the behaviour of the target insect. C. pomonella larvae do not ingest food during penetration of the fruit. The larva bites pieces of the epidermis and deposits them without ingestion on top of the entry hole.

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Zusammenfassung Das delta-Endotoxin von B.t. war in Laborversuch auf Kunstmedium gegenüber den Larven des Apfelwicklers, C. pomonella, sehr aktiv. Die hohe Aktivität konnte aber unter Feldbedingungen nicht mehr bestätigt werden.Es wurde nachgewiesen, dass die unbefriedigenden Resultate von B. thuringiensis unter Feldbedingungen auf das Verhalten der Junglarven zurückzuführen sind: Die Larven nehmen während dem Eindringen in den Apfel keine Nahrung auf, sondern deponieren die herausgebissenen Epidermistücke über der Einbohrstelle.

     

Infectivity of Echinostoma friedi miracidia to different snail species under experimental conditions

The infectivity of Echinostoma friedi (Trematoda: Echinostomatidae) miracidia was studied experimentally in a range of laboratory-reared snails that coexist in the same natural locality, namely Radix peregra, Lymnaea fuscus, L. truncatula (Lymnaeidae), Gyraulus chinensis, Helisoma duryi (Planorbidae) and Physella acuta (Physidae), and snails from different geographical origins acting naturally or experimentally as intermediate hosts of Schistosoma spp., namely Planorbarius metidjensis (from Málaga, Spain), Biomphalaria glabrata (Guadeloupe), B. alexandrina (Egypt) (Planorbidae), Bulinus cernicus (Mauritius), B. globosus (Zambia), B. natalensis (South Africa) and B. truncatus (Niger) (Bulinidae). Six species of snails were found to be susceptible, with the rate of infection ranging from 0 to 36.7%. The highest infection was detected in R. peregra. The low host specificity of E. friedi might have an epidemiological significance as a requisite for a recent establishment in a new geographical area.     

Nitrogen cycling in two riparian forest soils under different geomorphic conditions

At the floodplain scale, spatial pattern and successional development of riparian vegetation are under the control of geomorphic processes. The geomorphic and hydraulic characteristics of stream channels affect the sorting of organic material and inorganic sediment through erosion/sedimentation during floods. In turn, the proportion of fine sediments fractions differs by location within a given community of riparian forest succession. In this paper we tested the effect of geomorphic features of floodplains, through soil grain size sorting, on the nitrogen cycling in riparian forest soils. Two typical riparian forests formed by vertical accretion deposits from repeated addition of sediments from overbank flow have been chosen along the River Garonne, southwest France. These riparian forests had equivalent vegetation, flood frequency and duration, differing only in soil grain size composition: one riparian forest had sandy soils and the other had loamy soils. The evolution of the main soil physical and chemical parameters as well as denitrification (DNT), N uptake (N_U) and mineralization (N_M) rates were measured monthly over a period of 13 months in the two study sites. The loamy riparian forest presented a better physical retention of suspended matter during floods. Moreover,in situ denitrification rates (DNT) and N uptake by plants (N_U) measured in the loamy riparian forest soils were significantly greater than in the sandy soils. Although DNT and N_U could be in competition for available nitrogen, the peak rates of these two processes did not occur at the same period of the year, N_U being more important during the dry season when DNT was minimum, while DNT rates were maximum following the spring floods. N retention by uptake (N_U) and loss by DNT represented together the equivalent of 32% of total organic nitrogen deposited during floods on the sandy riparian forest soils and 70% on the loamy ones. These significant differences between the two sites show that, at the landscape level, one should not estimate the rates of N_U and DNT, in riparian forests soils only on the basis of vegetation, but should take also into account the geomorphic features of the floodplain.     

Flowcell culture of Porphyromonas gingivalis biofilms under anaerobic conditions

We have developed an anaerobic biofilm culture system. The system is inexpensive, simple to use and, unlike an anaerobic glovebox, requires no dedicated space. As a test of the system, Porphyromonas gingivalis was cultured under low oxygen (1–2 ppm) and under anaerobic conditions (≤0.1 ppm O₂). In the presence of small amounts of oxygen, the organism attached and formed an initial biofilm over the course of 4 h, but the biofilm was unable to maintain its growth and had lost biomass after 18 h. Also, ambiguous results were obtained when the biofilm was stained with a viability stain. Under anaerobic conditions, the biofilm was able to continue growth — biomass was greater after 18 h than after 4 h, and the anaerobic biofilm had a less ambiguous staining pattern than did the low-O₂-grown biofilm.