Effectiveness of an ultraviolet-C disinfection system for reduction of healthcare-associated pathogens.

BACKGROUND: Healthcare-associated infections caused by multidrug-resistant (MDR) pathogens are significantly associated with increased mortality and morbidity. Environmental cleaning can reduce transmission of these pathogens but is often inadequate. Adjunctive methods are warranted to enhance the effectiveness of disinfection particularly in hospital settings where healthcare-associated infections are of major concern. METHODS: We conducted a study to examine the effectiveness of a mobile, automatic device, Hyper Light Disinfection Robot (model: Hyper Light P3), which utilized ultraviolet-C (UV-C) to kill MDR-Pseudomonas aeruginosa, MDR- Acinetobacter baumannii, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecium (VRE), Mycobacterium abscessus and Aspergillus fumigatus. The performance of this device in disinfecting hospital rooms previously admitted by patients harboring MRSA and VRE was also assessed. RESULTS: Except for VRE and M. abscessus, more than 3 log10 reduction of vegetative bacteria colonies was observed after UV-C irradiation of 5 min at a distance of 3 m from the device. At the distance of 1 m, substantial and comparable reduction of colonies was observed across all tested microorganisms regardless of exposure time. The killing effect was less pronounced for A. fumigatus particularly at the distance of 2-3 m. In uncleaned hospital rooms, there was significant reduction in the number of bacteria colonies sampled from different surfaces after UV-C irradiation for 15 min. CONCLUSIONS: UV-C disinfection system was effective in killing MDR pathogens. Further study is warranted to confirm its effectiveness as an adjunctive method in disinfecting hospital environment.

Gamma radiation-induced in vitro hormetic apogamy in the fern Pityrogramma calomelanos (L.) link.

Pityrogramma calomelanos (L.) Link, popularly known as "Silver fern" has significant importance as a medicinal plant used traditionally for its astringent, analgesic, anti-haemorrhagic, anti-hypertensive, anti-pyretic and anthelminthic properties. This fern demonstrates an increased morphogenetic potential towards sporophyte formation, upon exposure to low doses of gamma radiation. Young sporophytic leaf crosier cultures were established in vitro on agar based Knop's media with and without 20 g/l sucrose. The cultures were subjected to 60Co radiations in the range of 2.5-100 Gy. Apospory (production of gametophytes on sporophytic tissue without spores) was observed on leaf tissue cultured on Knops media with and without sucrose in P. calomelanos, at the end of 60 days. 5 Gy treated explants showed high number of aposporous gametophytes and was comparable to the control. Other tested doses reduced the aposporous gametophyte production significantly. In the second phase of the experimentation, the cultures were retained on the gametophyte induction media for a period of 4 weeks. Aposporous gametophytes were observed to proliferate with occasional development of antheridia. At the end of 4 weeks, morphogenetic development on the gametophytic tissue resulted in a significantly higher number of apogamous sporophytes (production of sporophytes without fusion of gametes) were obtained on 5 Gy treated tissue as compared to control and all the other treated explants. Apogamous sporophytes thus produced were successfully grown in the greenhouse and transferred to the field. Thus the use of gamma radiation in vitro not only reduced the need for sucrose for induction of apospory in P.calomelanos, it also exhibited hormesis at 5 Gy for improved sporophyte production.

Spores potentially dispersed to longer distances are more tolerant to ultraviolet radiation: A case study in the moss genus Orthotrichum.

PREMISE OF THE STUDY: Ultraviolet (UV) radiation influences the viability of algal spores and seed-plant pollen depending on the species, the dose, and the wavelength. In bryophytes, one of the dominant groups of plants in many habitats, UV radiation could determine their spore dispersal strategy, and such data are critical for reconstructing the ancestral state in plants and for determining the distribution range and persistence of bryophyte species. METHODS: Spores of four bryophyte species of the moss genus Orthotrichum that were either hygrochastic or xerochastic (spores dispersed under wet or dry conditions, respectively) were exposed to realistic doses of UV radiation under laboratory conditions. Spore viability was evaluated through germination experiments and, for the first time in bryophytes, ultrastructural observations. Given that the UV-B doses used were relatively higher than the UV-A doses, the UV effect was probably due more to UV-B than UV-A wavelengths. KEY RESULTS: All four species reduced their spore germination capacity in a UV dose-dependent manner, concomitantly increasing spore ultrastructural damage (cytoplasmic and plastid alterations). Most spores eventually died when exposed to the highest UV dose. Interestingly, spores of hygrochastic species were much more UV-sensitive than those of xerochastic species. CONCLUSIONS: UV tolerance determines moss spore viability, as indicated by germination capacity and ultrastructural damage, and differs between spores of species with different dispersal strategies. Specifically, the higher UV tolerance of xerochastic spores may enable them to be dispersed to longer distances than hygrochastic spores, thus extending more efficiently the distribution range of the corresponding species.

Drinking water treatment with ultraviolet light for travelers -- Evaluation of a mobile lightweight system.

BACKGROUND: The SteriPEN(®) is a handheld device for disinfecting water with ultraviolet (UV) radiation. The manufacturer claims a reduction of at least 99.9% of bacteria, viruses, and protozoa. The present study intends to verify the general effectiveness of the device. Furthermore, the influence of bottle geometry and water movement is examined and the issue of user safety with regard to UV-C radiation is addressed. METHODS: The device was applied on water containing a known number of microorganisms (Escherichia coli, Staphylococcus aureus, and the spore of Geobacillusstearothermophilus) and the survival rate was examined. Three different types of bottles commonly used among travelers served as test containers. All tests were conducted with and without agitating the water during irradiation. Furthermore, a spectral analysis was performed on the light of the device. RESULTS: The SteriPEN(®) reached a mean reduction of more than 99.99% of bacteria and 99.57% of the spores when applied correctly. However, the results of the trials without agitating the water only yielded a 94.98% germ reduction. The device's maximal radiation intensity lies at 254 nm which is the wavelength most efficient in inactivating bacteria. The UV-C fraction is filtered out completely by common bottle materials. However, when applied in larger containers a portion of the UV-C rays exits the water surface. CONCLUSIONS: If applied according to the instructions the device manages a satisfactory inactivation of bacteria. However, it bears the danger of user errors relevant to health. Therefore, education on the risks of incorrect application should be included in the travel medical consultation. Also there are still aspects that need to be subject to further independent research.

The molecular and physiological responses of Physcomitrella patens to ultraviolet-B radiation.

Ultraviolet-B (UV-B) radiation present in sunlight is an important trigger of photomorphogenic acclimation and stress responses in sessile land plants. Although numerous moss species grow in unshaded habitats, our understanding of their UV-B responses is very limited. The genome of the model moss Physcomitrella patens, which grows in sun-exposed open areas, encodes signaling and metabolic components that are implicated in the UV-B response in flowering plants. In this study, we describe the response of P. patens to UV-B radiation at the morphological and molecular levels. We find that P. patens is more capable of surviving UV-B stress than Arabidopsis (Arabidopsis thaliana) and describe the differential expression of approximately 400 moss genes in response to UV-B radiation. A comparative analysis of the UV-B response in P. patens and Arabidopsis reveals both distinct and conserved pathways.

Effects of ultraviolet radiation and temperature on the ultrastructure of zoospores of the brown macroalga Laminaria hyperborea.

The interactive effects of an 8 h exposure to UV radiation and altered temperatures on the ultrastructure and germination of zoospores of the sublittoral brown alga Laminaria hyperborea (Gunn.) Foslie were investigated for the first time. Spores were exposed to four temperatures (2, 7, 12 and 17 degrees C) and three light regimes (PAR, PAR + UV-A, PAR + UV-A+UV-B). Freshly-released spores of L. hyperborea lack a cell wall and contain a nucleus with fine granular nucleoplasm and a nucleolus, one chloroplast, several mitochondria, dictyosomes and an endoplasmatic reticulum. Further, several kinds of so-called adhesive vesicles, lipid globuli and physodes containing UV-absorbing phlorotannins are embedded in the cytoplasm. No eye-spot is present. Physodes were found but they were rare and small. After an 8 h exposure to UV-B, the nucleoplasm had a mottled structure, chloroplasts contained plastoglobuli, the structure of the mitochondria changed from crista- to sacculus-type and germination was strongly inhibited at all temperatures. UV-A only had an impact on the ultrastructure at the highest temperature tested. The strongest effects were found at 17 degrees C, where germination was reduced to 35%, 32% and 9% after exposure to PAR, PAR+UV-A and PAR + UV-A + UV-B, respectively. This study indicates that UV-B radiation has strong damaging effects on the physiology and ultrastructure of zoospores of L. hyperborea. The results are important for developing scenarios for the effect of enhanced UV radiation and increasing temperatures caused by global climate changes.

Efficiency of Artemia cysts removal as a model invasive spore using a continuous microwave system with heat recovery.

A continuous microwave system to treat ballast water inoculated with Artemia salina cysts as a model invasive spore was tested for its efficacy in inactivating the cysts present. The system was tested at two different flow rates (1 and 2 L x min(-1)) and two different power levels (2.5 and 4.5 kW). Temperature profiles indicate that the system could deliver heating loads in excess of 100 degrees C in a uniform and near-instantaneous manner when using a heat recovery system. Except for a power and flow rate combination of 2.5 kW and 2 L x min(-1), complete inactivation of the cysts was observed at all combinations at holding times below 100 s. The microwave treatment was better or equal to the control treatment in inactivating the cysts. Use of heat exchangers increased the power conversion efficiency and the overall efficiency of the treatment system. Cost economics analysis indicates that in the present form of development microwave treatment costs are higher than the existing ballast water treatment methods. Overall, tests results indicated that microwave treatment of ballast water is a promising method that can be used in conjunction with other methods to form an efficient treatment system that can prevent introduction of potentially invasive spore forming species in non-native waters.

Rapid determination of spore chemistry using thermochemolysis gas chromatography-mass spectrometry and micro-Fourier transform infrared spectroscopy.

Spore chemistry is at the centre of investigations aimed at producing a proxy record of harmful ultraviolet radiation (UV-B) through time. A biochemical proxy is essential owing to an absence of long-term (century or more) instrumental records. Spore cell material contains UV-B absorbing compounds that appear to be synthesised in variable amounts dependent on the ambient UV-B flux. To facilitate these investigations we have developed a rapid method for detecting variations in spore chemistry using combined thermochemolysis gas chromatography-mass spectrometry and micro-Fourier transform infrared spectroscopy. Our method was tested using spores obtained from five populations of the tropical lycopsid Lycopodium cernuum growing across an altitudinal gradient (650-1981 m a.s.l.) in S.E. Asia with the assumption that they experienced a range of UV-B radiation doses. Thermochemolysis and subsequent pyrolysis liberated UV-B pigments (ferulic and para-coumaric acid) from the spores. All of the aromatic compounds liberated from spores by thermochemolysis and pyrolysis were active in UV-B protection. The various functional groups associated with UV-B protecting pigments were rapidly detected by micro-FTIR and included the aromatic C[double bond, length as m-dash]C absorption band which was exclusive to the pigments. We show increases in micro-FTIR aromatic absorption (1510 cm(-1)) with altitude that may reflect a chemical response to higher UV-B flux. Our results indicate that rapid chemical analyses of historical spore samples could provide a record ideally suited to investigations of a proxy for stratospheric O3 layer variability and UV-B flux over historical (century to millennia) timescales.

UV effects on photosynthesis and DNA in propagules of three Antarctic seaweeds (Adenocystis utricularis, Monostroma hariotii and Porphyra endiviifolium).

Ozone depletion is highest during spring and summer in Antarctica, coinciding with the seasonal reproduction of most macroalgae. Propagules are the life-stage of an alga most susceptible to environmental perturbations therefore, reproductive cells of three intertidal macroalgal species Adenocystis utricularis (Bory) Skottsberg, Monostroma hariotii Gain, and Porphyra endiviifolium (A and E Gepp) Chamberlain were exposed to photosynthetically active radiation (PAR), PAR + UV-A and PAR + UV-A + UV-B radiation in the laboratory. During 1, 2, 4, and 8 h of exposure and after 48 h of recovery, photosynthetic efficiency, and DNA damage were determined. Saturation irradiance of freshly released propagules varied between 33 and 83 mumol photons m(-2) s(-1) with lowest values in P. endiviifolium and highest values in M. hariotii. Exposure to 22 mumol photons m(-2) s(-1 )PAR significantly reduced photosynthetic efficiency in P. endiviifolium and M. hariotii, but not in A. utricularis. UV radiation (UVR) further decreased the photosynthetic efficiency in all species but all propagules recovered completely after 48 h. DNA damage was minimal or not existing. Repeated exposure of A. utricularis spores to 4 h of UVR daily did not show any acclimation of photosynthesis to UVR but fully recovered after 20 h. UVR effects on photosynthesis are shown to be species-specific. Among the tested species, A. utricularis propagules were the most light adapted. Propagules obviously possess good repair and protective mechanisms. Our study indicates that the applied UV dose has no long-lasting negative effects on the propagules, a precondition for the ecological success of macroalgal species in the intertidal.

Exposure to ultraviolet radiation delays photosynthetic recovery in Arctic kelp zoospores.

Seasonal reproduction in some Arctic Laminariales coincides with increased UV-B radiation due to stratospheric ozone depletion and relatively high water temperatures during polar spring. To find out the capacity to cope with different spectral irradiance, the kinetics of photosynthetic recovery was investigated in zoospores of four Arctic species of the order Laminariales, the kelps Saccorhiza dermatodea, Alaria esculenta, Laminaria digitata, and Laminaria saccharina. The physiology of light harvesting, changes in photosynthetic efficiency and kinetics of photosynthetic recovery were measured by in vivo fluorescence changes of Photosystem II (PSII). Saturation irradiance of freshly released spores showed minimal I ( k ) values (photon fluence rate where initial slope intersects horizontal asymptote of the curve) values ranging from 13 to 18 micromol photons m(-2) s(-1) among species collected at different depths, confirming that spores are low-light adapted. Exposure to different radiation spectra consisting of photosynthetically active radiation (PAR; 400-700 nm), PAR+UV-A radiation (UV-A; 320-400 nm), and PAR+ UV-A+UV-B radiation (UV-B; 280-320 nm) showed that the cumulative effects of increasing PAR fluence and the additional effect of UV-A and UV-B radiations on photoinhibition of photosynthesis are species specific. After long exposures, Laminaria saccharina was more sensitive to the different light treatments than the other three species investigated. Kinetics of recovery in zoospores showed a fast phase in S. dermatodea, which indicates a reduction of the photoprotective process while a slow phase in L. saccharina indicates recovery from severe photodamage. This first attempt to study photoinhibition and kinetics of recovery in zoospores showed that zoospores are the stage in the life history of seaweeds most susceptible to light stress and that ultraviolet radiation (UVR) effectively delays photosynthetic recovery. The viability of spores is important on the recruitment of the gametophytic and sporophytic life stages. The impact of UVR on the zoospores is related to the vertical depth distribution of the large sporophytes in the field.

Monitoring of solar-UV exposure among schoolchildren in five Japanese cities using spore dosimeter and UV-coloring labels.

To monitor personal exposure to biologically effective solar-UV radiation, Bacillus subtilis spores on a membrane filter and UV-coloring labels were incorporated into a monitoring badge. The samples were covered with one of three types of filter sheet, dependent on the season, to reduce the amounts of exposure to measurable levels. Five fifth- or sixth-grade classes of primary schools, each consisting of 30-40 children, were chosen in northern (Sapporo), central (Tsukuba and Tokyo), and southern (Miyazaki and Naha) cities in Japan. In all four season, each child wore a badge on an upper arm for the entire waking hours, changing it daily, for a week. Upon collection of the badges, the survival of spores and the extent of coloration of the label were determined. The results were used to estimate the amount of daily exposure to biologically effective UV radiation, expressed as the value of spore inactivation dose. Unexpectedly, the average amounts of exposure were not directly correlated with the outdoor UV irradiance: in the two southern cities, despite high outdoor irradiance from spring to autumn, the average amounts of exposure were less than 3.1% of the average irradiance. Highly concentrated exposures occurred in two central cities on three days when extensive outdoor exercise took place. These results contradict the simple notion that children's exposure is in proportion to the outdoor UV irradiance, and support the view that the extent of solar-UV exposure is primarily determined by life-style rather than living location.

Caffeine-resistance in S. pombe: mutations in three novel caf genes increase caffeine tolerance and affect radiation sensitivity, fertility, and cell cycle.

Caffeine is a well known base analogue and is cytotoxic to both animal and yeast cells. There are two possible mechanisms by which yeast cells tolerate caffeine concentrations higher than normal, by mutation or by physiological adaptation. We have isolated novel caffeine-resistant mutants of S. pombe which define three distinct genes caf2, caf3 and caf4. These mutants achieved a level of caffeine resistance which is presumed to represent the upper limit attainable by mutation. The caf2-caf4 mutations, as well as the previously identified caf1 mutation, confer UV-sensitivity, caffeine-resistant UV repair, impaired fertility and sporulation, as well as a lengthened cell cycle. They are partially dominant for caffeine resistance and recessive for UV sensitivity. Some auxotrophic caf3-89 double mutants show drastically decreased caffeine resistance. The caf4 mutant is more resistant to gamma-radiation than wild-type cells and shows pH-sensitive growth. As each caf mutation can, individually, confer maximum caffeine resistance to the cells, all four genes are expected to operate in the same pathway. This pathway might also be responsible for the physiological adaptation since adaptation is lost in caf1-caf4 mutants.

The effect of ultraviolet radiation on the germination of Nosema algerae Vávra and Undeen (Microsporida: Nosematidae) spores.

Spores of Nosema algerae Vávra and Undeen were subjected to various dosages of 254 nm ultraviolet radiation (UV). Very high dosages of UV were required to block germination. Germination was normal immediately after UV dosages of 0.2 to 1.0 J/cm2, followed by a delayed effect in which both percentage germination and the intrasporal concentration of trehalose decreased with time after UV exposure. Although a few spores were germinated, most of them were inactivated (rendered temporarily unable to germinate) by exposure to UV of 1.1 J/cm2. Ultraviolet radiation between 1.1 and 3.4 J/cm2 stimulated spores to germinate. However, spores were completely unable to germinate immediately after exposure to dosages above 3.8 J/cm2. Ammonia had little effect on stimulation by UV but was inhibitory to germination after stimulation had occurred. These results demonstrate that UV behaves like a germination stimulus and are discussed in terms of the hypothesis that germination is initiated by the breakdown of barriers between trehalose and trehalase.

The effects of heat and cobalt-60 gamma-radiation on excystation of the rat coccidium, Eimeria nieschulzi Dieben, 1924.

Sporulated oocysts of Eimeria nieschulzi Dieben, a rat coccidium, were exposed for 1 hr to Cobalt-60 gamma-radiation (15, 30, or 60k-rads), to heat (35, 40, or 45 C) , or to both concurrently (15, 30, OR 60 K-RADS AT 35 C) to compared the excystation capabilities of treated vs nontreated parasites. Intact treated oocysts appeared structurally unaltered when viewed with the light microscope. Excystation of sporozoites occured in all treated groups when their sporocysts were exposed to a trypsin-sodium taurocholate (TST) fluid, but after 150 min in TST the excystation rate was significantly lower than in nontreated sporocysts. Sporozoites which excysted from treated sporocysts were abnormal both in the excystation process and in their form and movement once outside the sporocysts.

Effects of ionizing radiation on dried spores of Osmunda regalis III. 35 GHz e.s.r. study.

The use of 35 GHz microwave frequency has greatly simplified the separation of overlapping signals in dried irradiated fern spores. While in general confirming the conclusions previously drawn from X-band spectra, the use of Q-band has clarified several assignments of radical groups and allowed the recognition of additional radical groups or sub-groups.

Effects of ionizing radiation on dried spores of Osmunda regalis III. 35 GHz e.s.r. study.

The use of 35 GHz microwave frequency has greatly simplified the separation of overlapping signals in dried irradiated fern spores. While in general confirming the conclusions previously drawn from X-band spectra, the use of Q-band has clarified several assignments of radical groups and allowed the recognition of additional radical groups or sub-groups.