Comparison of UV Inactivation of Spores of Three Encephalitozoon Species with That of Spores of Two DNA Repair-Deficient Bacillus subtilis Biodosimetry Strains

When exposed to 254-nm UV, spores of Encephalitozoon intestinalis, Encephalitozoon cuniculi, and Encephalitozoon hellem exhibited 3.2-log reductions in viability at UV fluences of 60, 140, and 190 J/m², respectively, and demonstrated UV inactivation kinetics similar to those observed for endospores of DNA repair-defective mutant Bacillus subtilis strains used as biodosimetry surrogates. The results indicate that spores of Encephalitozoon spp. are readily inactivated at low UV fluences and that spores of UV-sensitive B. subtilis strains can be useful surrogates in evaluating UV reactor performance.     

Genetic and enzymatic analysis of a gene controlling UV sensitivity in Neurospora crassa

The upr-I gene controlling UV sensitivity in Neurospora crassa has been transferred from a microconidial strain into a macroconidial strain. The properties of the strain with respect to UV sensitivity and photoreactivation (PR) have been modified in the macroconidial background leading to an enhancement of resistance to UV by a factor of about 2 and some suggestion of enzymatic PR. The upr-I gene confers sensitivity to nitrous acid to about the same degree as that for UV. However, inactivation of strains carrying the upr-I gene by white light in the presence of photosensitizing dyes is equivalent to that of strains exhibiting normal sensitivity to UV.     

Male attractiveness is influenced by UV wavelengths in a newt species but not in its close relative

Background

Functional communication in the UV range has been reported in Invertebrates and all major groups of Vertebrates but Amphibians. Although perception in this wavelength range has been shown in a few species, UV signalling has not been demonstrated in this group. One reason may be that in lentic freshwater habitats, litter decomposition generates dissolved organic carbon that absorbs UV radiation and thus hinders its use for visual signalling. We tested the effect of male UV characteristics on female sexual preference in two newt species that experience contrasting levels of UV water transmission when breeding.

Methodology/Principal Findings

We analysed water spectral characteristics of a sample of breeding ponds in both species. We quantified male ventral coloration and measured male attractiveness under two lighting conditions (UV present, UV absent) using a no-choice female preference design. UV transmission was higher in Lissotriton vulgaris breeding sites. Male UV patterns also differed between experimental males of the two species. We observed a first common peak around 333 nm, higher in L. vulgaris, and a second peak around 397 nm, more frequent and higher in L. helveticus. Male attractiveness was significantly reduced in L. vulgaris when UV was not available but not in L. helveticus. Male attractiveness depended on the hue of the first UV peak in L. vulgaris.

Conclusion/Significance

Our study is the first report of functional UV-based communication in Amphibians. Interestingly, male spectral characteristics and female preferences were consistent with the differences in habitat observed between the two species as L. helveticus often breeds in ponds containing more UV blocking compounds. We discuss the three hypotheses proposed so far for UV signalling in animals (enhanced signal detectability, private communication channel, indicator of individual quality).     

UV Inactivation of Cryptosporidium hominis as Measured in Cell Culture

The Cryptosporidium spp. UV disinfection studies conducted to date have used Cryptosporidium parvum oocysts. However, Cryptosporidium hominis predominates in human cryptosporidiosis infections, so there is a critical need to assess the efficacy of UV disinfection of C. hominis. This study utilized cell culture-based methods to demonstrate that C. hominis oocysts displayed similar levels of infectivity and had the same sensitivity to UV light as C. parvum. Therefore, the water industry can be confident about extrapolating C. parvum UV disinfection data to C. hominis oocysts.     

Immunohistochemical evidence of cone-based ultraviolet vision in divergent bat species and implications for its evolution

We characterized Fos-like expression patterns in the primary visual cortex (V1) by binocular flicking stimulation with UV light to investigate cone-based UV vision in four bat species representing four lineages: Hipposideros armiger and Scotophilus kuhlii, insectivores using constant frequency (CF) or frequency modulation (FM) echolocation, respectively, and Rousettus leschenaultii and Cynopterus sphinx, cave-roosting and tree-roosting fruit bats, respectively. The optic centre processing the visual image, V1, appears more distinctly immunostaining in S. kuhlii and C. sphinx after 1 h of UV light stimuli while in H. armiger and R. leschenaultii, staining was no more distinct than in corresponding controls. Our immunohistochemical evidence supports differences in the distribution of cone-based UV vision in the order Chiroptera and supports our earlier postulate that due to possible sensory tradeoffs and roosting ecology, defects in the short wavelength opsin genes have resulted in loss of UV vision in CF but not in FM bats. In addition, fruit bats roosting in caves have lost UV vision but not those roosting in trees. Our results thus confirm that bats are a further mammalian taxon that has retained cone-based UV sensitivity in some species.     

Partial suppression of the LexA phenotype by mutations (rnm) which restore ultraviolet resistance but not ultraviolet mulability to Escherichia coli B/r uvrA lexA

In Escherichia coli, lexA mutations eliminate expression of UV-inducible functions, causing pleiotropic effects which include sensitivity to ultraviolet (UV) light and loss of UV mutability. Selection for UV resistance, after 5-bromouracil (BU) treatment of E. coli B/r uvr A lexA-102, has yielded derivatives more resistant than lexA but still refractory to UV mutagenesis. The mutation responsible for the UV-resistant UV-nonmutable phenotype (rnm) is cotransducible with malB to about the same extent as is lexA-102 and is tightly linked to lexA-102 in at least one strain. The rnm mutation may therefore be an intragenic partial suppressor of the LexA phenotype. In addition to increased UV resistance and lack of UV mutability, rnm strains show improved ability to perform postreplication repair and to control postirradiation DNA degration compared to the lexA parent. We ascribe the properties of rnm mutants to their having reacquired control of Exonuclease V activity without having reacquired UV-inducible error-prone postreplication repair. We relate our results to current interpretations of UV mutagenesis and to models of coordinate regulation of UV-inducible functions.     

A New UV-A/B Protecting Pigment in the Terrestrial Cyanobacterium Nostoc commune

A new ultraviolet (UV)-A/B absorbing pigment with maxima at 312 and 330 nanometers from the cosmopolitan terrestrial cyanobacterium Nostoc commune is described. The pigment is found in high amounts (up to 10% of dry weight) in colonies grown under solar UV radiation but only in low concentrations in laboratory cultures illuminated by artificial light without UV. Its experimental induction by UV as well as its capacity to efficiently protect Nostoc against UV radiation is reported.     

Deinococcus radiodurans UV endonuclease β DNA incisions do not generate photoreversible thymine residues

The ability of UV endonuclease β of Deinococcus radiodurans to act as a pyrimidine dimer DNA glycosylase was investigated. Cell-free extracts of D. radiodurans exhibiting UV endonuclease β activity failed to generate incisions in irradiated DNA that liberated free-thymine residues upon photoreversal with 254-nm light. This is in marked contrast to the pyrimidine dimer UV glycosylase of Micrococcus luteus that does liberate such residues. The result suggests that UV endonuclease β incises DNA by true endonuclease action.     

The synergism between hydrocarbon pollutants and UV radiation: a potential link between coastal pollution and larval mortality

Coastal, benthic invertebrates with complex life history strategies are exposed to stage- and habitat-specific selective forces. In the coastal environment, benthic adults are exposed to polycyclic aromatic hydrocarbon pollutants (PAHs) due to their proximity to human activities (shipping, urbanization, and industrialization). Benthic invertebrates produce lipid-rich eggs or larvae that absorb PAHs from polluted estuaries and coastal waters. The larvae of many coastal invertebrates move offshore following release from benthic adults. During development in offshore waters, larvae of some species are exposed to relatively high levels of ultraviolet (UV) radiation. Marine organisms vary in their tolerance to PAHs and UV radiation. The purpose of this study was to examine the effects of the sequential exposure of the larvae of marine crabs to PAHs and UV radiation.Using laboratory experiments, the larvae of four crab species were exposed to PAHs and UV radiation. There was a significant synergistic effect of exposure to PAH (fluoranthene or pyrene) and UV radiation on larvae of the spider crab (Libinia dubia), the stone crab (Menippe adina) and the mud crab (Panopeus herbstii). Larvae of blue crabs (Callinectes sapidus) were exposed to PAHs and UV radiation in both laboratory and solar UV experiments. Significantly higher mortality occurred for C. sapidus larvae using either type of UV-artificial or solar.Larvae of coastal invertebrates with complex life history strategies are susceptible to the combined effects of PAHs and UV radiation. In this study, the exposure of crab larvae to PAHs and UV radiation resulted in mortality to crab larvae using laboratory and solar UV experiments. There were no effects on larval crab mortality due to PAH or UV radiation independently but mortality was as high as 100% when both factors were present.     

Induction of anthocyanin formation and of enzymes related to its biosynthesis by UV light in cell cultures of Haplopappus gracilis

Only UV light below 345 nm stimulates anthocyanin formation in dark grown cell suspension cultures of Haplopappus gracilis. A linear relationship between UV dose and flavonoid accumulation, as found previously with parsley cell cultures, was not observed with the H. gracilis cells. Only continuous irradiation with high doses of UV was effective. Drastic increases in the activities of the enzymes phenylalanine ammonia-lyase, chalcone isomerase and flavanone synthase were observed under continuous UV light. The increase in enzyme activities paralleled anthocyanin formation.     

UV-A/B radiation rapidly activates photoprotective mechanisms in Chlamydomonas reinhardtii

Conversion of light energy into chemical energy through photosynthesis in the chloroplasts of photosynthetic organisms is essential for photoautotrophic growth, and non-photochemical quenching (NPQ) of excess light energy prevents the generation of reactive oxygen species and maintains efficient photosynthesis under high light. In the unicellular green alga Chlamydomonas reinhardtii, NPQ is activated as a photoprotective mechanism through wavelength-specific light signaling pathways mediated by the phototropin (blue light) and ultra-violet (UV) light photoreceptors, but the biological significance of photoprotection activation by light with different qualities remains poorly understood. Here, we demonstrate that NPQ-dependent photoprotection is activated more rapidly by UV than by visible light. We found that induction of gene expression and protein accumulation related to photoprotection was significantly faster and greater in magnitude under UV treatment compared with that under blue- or red-light treatment. Furthermore, the action spectrum of UV-dependent induction of photoprotective factors implied that C. reinhardtii senses relatively long-wavelength UV (including UV-A/B), whereas the model dicot plant Arabidopsis (Arabidopsis thaliana) preferentially senses relatively short-wavelength UV (mainly UV-B/C) for induction of photoprotective responses. Therefore, we hypothesize that C. reinhardtii developed a UV response distinct from that of land plants.

In contrast to land plants, which sense short-wave UV light, the unicellular green alga Chlamydomonas reinhardtii senses long-wavelength UV light for photoprotective responses.     

Studies on the ultraviolet light sensitivity of Bloom's syndrome fibroblasts

The sensitivity of Bloom's syndrome (bl/bl) fibroblasts to ultraviolet light (254 nm) has been estimated by 4 criteria: sister-chromatid exchange (SCE) formation, micronucleus production, cell survival, and host-cell reactivation of UV-irradiated adenovirus 2. In general, bl/bl strains did not differ significantly from the normal (+/+) strains in their response to UV treatment by any of the 4 criteria. One bl/bl strain, GM1492, was exceptional: It was abnormally sensitive to UV light in the SCE, micronucleus, and host-cell reactivation assays, but was not sensitive to UV as estimated by colony-forming ability. Thus, although one of the bl/bl strains studied in the experiments was sensitive to UV light as judged by some criteria, UV sensitivity is not a universal characteristic of Bloom's syndrome cells. It is nuclear whether the UV sensitivity of the GM1492 strain reflects genetic diversity within the syndrome or some unrelated property of this strain.     

Inactivation of Mycobacterium avium Complex by UV Irradiation

The effectiveness of two major UV technologies against a highly prevalent species of Mycobacterium avium complex was investigated. Our study indicates that M. avium is much more resistant to UV irradiation than most waterborne pathogens and that it is one of the rare microorganisms that are highly resistant to both chemical and UV disinfection in water.     

Phosphoinositide-specific phospholipase C forms a complex with 14-3-3 proteins and is involved in expression of UV resistance in fission yeast

The fission yeast plc1 ⁺ gene encodes phosphoinositide-specific phospholipase C. The two- hybrid interaction assay with plexA-plc1 ⁺ as a bait revealed that Plc1p interacted with the 14-3-3 proteins Rad24p and Rad25p. Formation of a complex containing Plc1p and Rad24p in vivo was confirmed by an immunological method. As predicted from the fact that rad24 null mutant cells are hypersensitive to UV irradiation, plc1 null mutant cells were almost as sensitive to UV irradiation as rad24 null mutant cells. In addition, deletion of rad24 in the plc1 null mutant cells did not enhance the UV sensitivity, indicating that plc1 ⁺ and rad24 ⁺ belong to the same epistasis group with respect to UV sensitivity. Whereas Rad24p has been reported to be involved in the DNA damage checkpoint pathway, the delay to mitosis after UV irradiation was not defective either in rad24 null mutant cells or in plc1 null mutant cells in our analysis. Thus, Plc1p is responsible for resistance to UV irradiation, but not for the DNA damage checkpoint pathway, in cooperation with 14-3-3 proteins.     

Radiation-sensitive pyrimidine auxotrophs of Ustilago Maydis II. A study of repair mechanisms and UV recovery in pyr 1

Two mutants at the pyr 1 locus have been used to study the radiation sensitivity of pyrimidine auxotrophs of U. maydis. The mutant pyr 1-1 has a reduced level of thymidine nucleotides, and this is a likely basis of the sensitivity. This strain is able to excise pyrimidine dimers from its DNA and is cross-sensitive to γ-rays and nitrosoguanidine (NG) as well as to UV. A diploid heteroallelic at the pyr 1 locus was UV-sensitive but not deficient in UV-induced mitotic recombination. The results suggest that the UV sensitivity may be due to the failure of a repair DNA polymerase to fill post-excision single-strand gaps in the DNA.The mutant pyr 1-1 exhibits the property of UV recovery, and this is shown to be dependent on the presence of dimers in the DNA. A mechanism for UV recovery is proposed in which a repair system, possibly involving recombination, is induced by the UV irradiation.     

Study of the Response of a Biofilm Bacterial Community to UV Radiation

We have developed a bioluminescent whole-cell biosensor that can be incorporated into biofilm ecosystems. RM4440 is a Pseudomonas aeruginosa FRD1 derivative that carries a plasmid-based recA-luxCDABE fusion. We immobilized RM4440 in an alginate matrix to simulate a biofilm, and we studied its response to UV radiation damage. The biofilm showed a protective property by physical shielding against UV C, UV B, and UV A. Absorption of UV light by the alginate matrix translated into a higher survival rate than observed with planktonic cells at similar input fluences. UV A was shown to be effectively blocked by the biofilm matrix and to have no detectable effects on cells contained in the biofilm. However, in the presence of photosensitizers (i.e., psoralen), UV A was effective in inducing light production and cell death. RM4440 has proved to be a useful tool to study microbial communities in a noninvasive manner.     

UV irradiation of poplar cells of Drosophila melanogaster embryos. V. A study of the meiotic recombination in females with chromosomes of different structure

Polar cells of female embryos heterozygous for the genetic markers st in ri were UV irradiated. The effect of UV irradiation on the meiotic recombination within the st-ri segment was analysed. In individuals that contained this chromosome segment in its normal position, UV increased the rate of meiotic recombination. If this segment were separated from the centromeric heterochromatin and translocated on the left arm of chromosome II, UV did not influence meiotic recombination to a significant extent. On the other hand, in non-irradiated controls the individuals containing the translocated segment showed an increased rate of recombination within the segment compared with individuals having the segment in its normal position.These results can be explained by Suzuki's hypothesis according to which a heterocatalytic activity of heterochromatin shows an antagonism to meiotic recombination. UV seems to interfere with this antagonism indirectly via induction of physiological damage.     

UV-induced DNA damage promotes resistance to the biotrophic pathogen Hyaloperonospora parasitica in Arabidopsis

Plant innate immunity to pathogenic microorganisms is activated in response to recognition of extracellular or intracellular pathogen molecules by transmembrane receptors or resistance proteins, respectively. The defense signaling pathways share components with those involved in plant responses to UV radiation, which can induce expression of plant genes important for pathogen resistance. Such intriguing links suggest that UV treatment might activate resistance to pathogens in normally susceptible host plants. Here, we demonstrate that pre-inoculative UV (254 nm) irradiation of Arabidopsis (Arabidopsis thaliana) susceptible to infection by the biotrophic oomycete Hyaloperonospora parasitica, the causative agent of downy mildew, induces dose- and time-dependent resistance to the pathogen detectable up to 7 d after UV exposure. Limiting repair of UV photoproducts by postirradiation incubation in the dark, or mutational inactivation of cyclobutane pyrimidine dimer photolyase, (6-4) photoproduct photolyase, or nucleotide excision repair increased the magnitude of UV-induced pathogen resistance. In the absence of treatment with 254-nm UV, plant nucleotide excision repair mutants also defective for cyclobutane pyrimidine dimer or (6-4) photoproduct photolyase displayed resistance to H. parasitica, partially attributable to short wavelength UV-B (280–320 nm) radiation emitted by incubator lights. These results indicate UV irradiation can initiate the development of resistance to H. parasitica in plants normally susceptible to the pathogen and point to a key role for UV-induced DNA damage. They also suggest UV treatment can circumvent the requirement for recognition of H. parasitica molecules by Arabidopsis proteins to activate an immune response.     

Changes in Phenolic Compounds and Cellular Ultrastructure of Arctic and Antarctic Strains of Zygnema (Zygnematophyceae, Streptophyta) after Exposure to Experimentally Enhanced UV to PAR Ratio

Ultraviolet (UV) radiation has become an important stress factor in polar regions due to anthropogenically induced ozone depletion. Although extensive research has been conducted on adaptations of polar organisms to this stress factor, few studies have focused on semi-terrestrial algae so far, in spite of their apparent vulnerability. This study investigates the effect of UV on two semi-terrestrial arctic strains (B, G) and one Antarctic strain (E) of the green alga Zygnema, isolated from Arctic and Antarctic habitats. Isolates of Zygnema were exposed to experimentally enhanced UV A and B (predominant UV A) to photosynthetic active radiation (PAR) ratio. The pigment content, photosynthetic performance and ultrastructure were studied by means of high-performance liquid chromatography (HPLC), chlorophyll a fluorescence and transmission electron microscopy (TEM). In addition, phylogenetic relationships of the investigated strains were characterised using rbcL sequences, which determined that the Antarctic isolate (E) and one of the Arctic isolates (B) were closely related, while G is a distinct lineage. The production of protective phenolic compounds was confirmed in all of the tested strains by HPLC analysis for both controls and UV-exposed samples. Moreover, in strain E, the content of phenolics increased significantly (p?=?0.001) after UV treatment. Simultaneously, the maximum quantum yield of photosystem II photochemistry significantly decreased in UV-exposed strains E and G (p?<?0.001), showing a clear stress response. The phenolics were most probably stored at the cell periphery in vacuoles and cytoplasmic bodies that appear as electron-dense particles when observed by TEM after high-pressure freeze fixation. While two strains reacted moderately on UV exposure in their ultrastructure, in strain G, damage was found in chloroplasts and mitochondria. Plastidal pigments and xanthophyll cycle pigments were investigated by HPLC analysis; UV A- and UV B-exposed samples had a higher deepoxidation state as controls, particularly evident in strain B. The results indicate that phenolics are involved in UV protection of Zygnema and also revealed different responses to UV stress across the three strains, suggesting that other protection mechanisms may be involved in these organisms.     

Inactivation of Salmonella enterica by UV-C Light Alone and in Combination with Mild Temperatures

The aim of this investigation was to study the efficacy of the combined processes of UV light and mild temperatures for the inactivation of Salmonella enterica subsp. enterica and to explore the mechanism of inactivation. The doses to inactivate the 99.99% (4D) of the initial population ranged from 18.03 (Salmonella enterica serovar Typhimurium STCC 878) to 12.75 J ml⁻¹ (Salmonella enterica serovar Enteritidis ATCC 13076). The pH and water activity of the treatment medium did not change the UV tolerance, but it decreased exponentially by increasing the absorption coefficient. An inactivating synergistic effect was observed by applying simultaneous UV light and heat treatment (UV-H). A less synergistic effect was observed by applying UV light first and heat subsequently. UV did not damage cell envelopes, but the number of injured cells was higher after a UV-H treatment than after heating. The synergistic effect observed by combining simultaneous UV and heat treatment opens the possibility to design combined treatments for pasteurization of liquid food with high UV absorptivity, such as fruit juices.