DNA-based confirmation that the parasitic wasp <Emphasis Type="Italic">Cotesia glomerata</Emphasis> (Braconidae,Hymenoptera) is a new threat to endemic butterflies of the Canary Islands

Island-endemic species can be particularly vulnerable to alien invasion. There are many examples of introduced insect parasitoids having a serious impact on endemic butterflies and moths. In 2006, a population of parasitic wasps was reared from larvae of the Canary Island Large White butterfly (Pieris cheiranthi), an endemic inhabitant of laurel forests unique to the Canary Islands of Macaronesia. Parasitoids were tentatively identified as Cotesia glomerata (Braconidae, Hymenoptera), a widely introduced agricultural bioagent. To corroborate this finding we sequenced 632 bp of mitochondrial cox1 from parasitoids and hosts from La Palma and from the native range of C. glomerata in continental Europe. These were combined with GenBank sequences and a character-based, phylogenetic approach was used to assess the species status of parasites and hosts. The La Palma parasitoid could unambiguously be assigned to C. glomerata under the criterion of diagnosibility with corroboration from multiple lines of evidence (DNA, morphology). We suggest that this opportunistic, non-native parasitoid wasp will be a threat to P. cheiranthi and other endemic Canarian butterflies. Parasitoid populations were recorded from P. cheiranthi in marginal forest habitats but not in central forest areas, suggesting that comprehensive habitat conservation of the Canarian laurel forests could prevent penetration of the alien parasitoid wasps and subsequent mortality of endemic butterfly populations.     

Carotenoid S(1) state in a recombinant light-harvesting complex of Photosystem II.

The carotenoid species lutein, violaxanthin, and zeaxanthin are crucial in the xanthophyll-dependent nonphotochemical quenching occurring in photosynthetic systems of higher plants, since they are involved in dissipation of excess energy and thus protect the photosynthetic machinery from irreversible inhibition. Nonetheless, important properties of the xanthophyll cycle carotenoids, such as the energy of their S(1) electronic states, are difficult to study and were only recently determined in organic solvents [Polívka, T. (1999) Proc. Natl. Acad. Sci. U.S.A. 96, 4914. Frank, H. A. (2000) Biochemistry 39, 2831]. In the present study, we have determined the S(1) energies of three carotenoid species, violaxanthin, lutein, and zeaxanthin, in their LHCII (peripheral light-harvesting complex of photosystem II) protein environment by constructing recombinant Lhcb1 (Lhc = light-harvesting complex) proteins containing single carotenoid species. Within experimental error the S(1) energy is the same for all three carotenoids in the monomeric LHCII, 13,900 +/- 300 cm(-1) (720 +/- 15 nm), thus well below the Q(y)() transitions of chlorophylls. In addition, we have found that, although the S(1) lifetimes of violaxanthin, lutein, and zeaxanthin differ substantially in solution, when incorporated into the LHCII protein, their S(1) states have in fact the same lifetime of about 11 ps. Despite the similar spectroscopic properties of the carotenoids bound to the LHCII, we observed a maximal fluorescence quenching when zeaxanthin was present in the LHCII complex. On the basis of these observations, we suggest that, rather than different photochemical properties of individual carotenoid species, changes in the protein conformation induced by binding of carotenoids with distinct molecular structures are involved in the quenching phenomena associated with Lhc proteins.     

Absence of co-phylogeny indicates repeated diatom capture in dinophytes hosting a tertiary endosymbiont

Tertiary endosymbiosis is proven through dinophytes, some of which (i.e. Kryptoperidiniaceae) have engulfed diatom algae containing a secondary plastid. Chloroplasts are usually inherited together permanently with the host cell, leading to co-phylogeny. We compiled a diatom sequence data matrix of two nuclear and two chloroplast loci. Almost all endosymbionts of Kryptoperidiniaceae found their closest relatives in free-living diatoms and not in other harboured algae, rejecting co-phylogeny and indicating that resident diatoms were taken up by dinophytes multiple times independently. Almost intact ultrastructure and insignificant genome reduction are supportive for young, if not recent events of diatom capture. With their selective specificity on the one hand and extraordinary degree of endosymbiotic flexibility on the other hand, dinophytes hosting diatoms share more traits with lichens or facultatively phototrophic ciliates than with green algae and land plants. Time estimates indicate the dinophyte lineages as consistently older than the hosted diatom lineages, thus also favouring a repeated uptake of endosymbionts. The complex ecological role of dinophytes employing a variety of organismic interactions may explain their high potential and plasticity in acquiring a great diversity of plastids.     

Time matters. Locomotor behavior of <Emphasis Type="Italic">Lacerta viridis</Emphasis> and <Emphasis Type="Italic">Lacerta agilis</Emphasis> in an open field maze

Locomotor performance provides one of the key pieces of information regarding whole-organism function. Experiments encompassing behavioral data commonly endeavor to measure parameters such as burst speed, latency time, distance traveled, and other aspects of locomotion. Behavioral experiments can uncover an immense range of information, from the individual, interspecific, and intraspecific levels up to correlations with ecological factors and parameters from the ecosystem. Here, we explored the locomotor behavior of two lizard species, Lacerta viridis and Lacerta agilis, in an open field test (OFT). The main aim was to reveal changes in locomotion over time. Although we observed no time-related variation in L. agilis, we discovered significant changes in locomotor behavior over the course of the experiment in Lacerta viridis. Measured behavioral traits (resting time, total distance traveled, mean speed) showed significant changes across time in L. viridis, thus indicating the importance of time as a factor when conducting behavioral experiments. Moreover, we observed that in 10-min experimental session, the individuals have undergone different stages from freezing behavior, exploration, to habituation.     

Relation to enterocins of variable <Emphasis Type="Italic">Aeromonas</Emphasis> species isolated from trouts of Slovakian aquatic sources and detected by MALDI-TOF mass spectrometry

Aeromonads represent bacteria thought to be primarily mostly autochthonous to aquatic environments. This study was focused on the relation with antibiotics and enterocins of identified Aeromonas species isolated from the intestine of trouts living in Slovakian aquatic sources. Intestinal samples from 50 trouts (3 Salmo trutta and 47 Salmo gairdnerii) were collected in April of years 2007, 2010, and 2015 from trouts of different water sources in Slovakia (pond Bukovec near Ko?ice, river ?ierny Váh). Due to the MALDI-TOF mass spectrometry evaluation, 25 strains were proposed to the genus Aeromonas involving nine different species (Aeromonas bestiarum—nine strains, Aer. salmonicida—four strains, Aer. encheleia, Aer. eucrenophila, Aer. molluscorum, Aer. media, Aer. sobria, Aer. popoffii, Aer. veronii). Phenotypic evaluation of individual strains confirmed their species identification. Twenty-five strains of different Aeromonas species were sensitive to azithromycin, amikacin, mecillinam, mezlocillin, piperacillin, gentamicin, chloramphenicol, and tetracycline. On the other side, they were resistant to carbenicillin and ticarcillin. The growth of Aer. bestiarum R41/1 was inhibited by treatment with Ent M and Ent 2019 (inhibition activity 100 AU/mL). Aer. bestiarum R47/3 was inhibited by eight enterocins (100 AU/mL). It is the first study testing enterocins to inhibit the growth of Aeromonas species from trouts.     

Aphids (Hemiptera: Aphidoidea) on cultivated and self-sown plants in Greece

The present work covers original information on aphids (Hemiptera: Aphidoidea) from Greece during 1995–2005. The individual aphid taxa were collected from their host plants and identified. The plant taxa were classified in two major categories: cultivated and self-sown. The cultivated ones were classified as: herbaceous, climbing and trees. The selfsown ones were classified as: annual herbaceous, annual or biennial herbaceous, biennial or perennial herbaceous, perennial herbaceous, shrubby and trees Two hundred twelve plant taxa were identified on which 105 aphid taxa were recorded. We report 391 plant taxa-aphid taxa associations from which, 190 are referred to cultivated and 201 to self-shown plant taxa. Aphis gossypii was found to infest the widest spectrum of cultivated plant taxa whereas Brachycaudus helichrysi was found to infest the widest spectrum of self-sown plant taxa. Myzus persicae was found to infest the widest spectrum of the total number of plant taxa. Information for several economically unimportant aphid taxa in the area are also presented.     

Effectiveness of mowing for the flora diversity preservation: a case study of steppe-like grassland in Croatia (NATURA 2000 site)

Intensive human activities have resulted in a critical reduction of grasslands in Croatia, one of the richest Europian countries in terms of biodiversity. Sub-Pannonic steppic grasslands are very rare, only few occur in the continental part of Croatia and most of them are protected as NATURA 2000 sites. We studied vascular flora of the Sub-Pannonic stepic grassland with the aim to examine the changes in plant communities 15 years from the mowing application. Results showed that hand mowing once per year at the end of vegetation season increased the species diversity as well as the number of medium and low growth taxa. The critically endangered, nearly threatened and vulnerable species remained preserved. The occurrence of trees, shrubs and invasive species as a consequence of succession and anthropogenic influences could be important threat for steppe-like grassland flora. From a conservation point of view infrequent mowing regime could be an effective management tool, although it needs to be adapted to regional and local circumstances for maintaining high biodiversity of steppe-like grasslands and valuable plant species.     

Optical microscopy in photosynthesis

Emerging as well as the most frequently used optical microscopy techniques are reviewed and image contrast generation methods in a microscope are presented, focusing on the nonlinear contrasts such as harmonic generation and multiphoton excitation fluorescence. Nonlinear microscopy presents numerous advantages over linear microscopy techniques including improved deep tissue imaging, optical sectioning, and imaging of live unstained samples. Nonetheless, with the exception of multiphoton excitation fluorescence, nonlinear microscopy is in its infancy, lacking protocols, users and applications; hence, this review focuses on the potential of nonlinear microscopy for studying photosynthetic organisms. Examples of nonlinear microscopic imaging are presented including isolated light-harvesting antenna complexes from higher plants, starch granules, chloroplasts, unicellular alga Chlamydomonas reinhardtii, and cyanobacteria Leptolyngbya sp. and Anabaena sp. While focusing on nonlinear microscopy techniques, second and third harmonic generation and multiphoton excitation fluorescence microscopy, other emerging nonlinear imaging modalities are described and several linear optical microscopy techniques are reviewed in order to clearly describe their capabilities and to highlight the advantages of nonlinear microscopy.     

Immunomodulatory strategies prevent the development of autoimmune emphysema

Background

The presence of anti-endothelial cell antibodies and pathogenic T cells may reflect an autoimmune component in the pathogenesis of emphysema. Whether immune modulatory strategies can protect against the development of emphysema is not known.

Methods

Sprague Dawley rats were immunized with human umbilical vein endothelial cells (HUVEC) to induce autoimmune emphysema and treated with intrathymic HUVEC-injection and pristane. Measurements of alveolar airspace enlargement, cytokine levels, immuno histochemical, western blot analysis, and T cell repertoire of the lung tissue were performed.

Results

The immunomodulatory strategies protected lungs against cell death as demonstrated by reduced numbers of TUNEL and active caspase-3 positive cells and reduced levels of active caspase-3, when compared with lungs from HUVEC-immunized rats. Immunomodulatory strategies also suppressed anti-endothelial antibody production and preserved CNTF, IL-1alpha and VEGF levels. The immune deviation effects of the intrathymic HUVEC-injection were associated with an expansion of CD4+CD25+Foxp3+ regulatory T cells. Pristane treatment decreased the proportion of T cells expressing receptor beta-chain, Vβ16.1 in the lung tissue.

Conclusions

Our data demonstrate that interventions classically employed to induce central T cell tolerance (thymic inoculation of antigen) or to activate innate immune responses (pristane treatment) can prevent the development of autoimmune emphysema.