Tolfenamic acid degradation by direct photolysis and the UV-ABC/H2O2 process: factorial design, kinetics, identification of intermediates, and toxicity evaluation.

This study employed direct UV-ABC photolysis and the UV-ABC/H2O2 process to investigate the degradation of tolfenamic acid (TA), a common anti-inflammatory drug used in both human and veterinary medicine. A 23 factorial design with added center point was used to evaluate the effect of three independent variables-namely, H2O2 concentration ([H2O2]), TA concentration ([TA]), and experiment time (time)-on TA degradation and H2O2 photolysis during UV-ABC/H2O2 treatment using a high-pressure mercury vapor lamp (photon flux of 2.6307 × 104 J s-1) as the UV irradiation source. The responses yielded similar values, revealing a linear behavior, with correlation coefficients R = 0.9968 and Radj = 0.9921 for TA degradation and R = 0.9828 and Radj = 0.9570 for H2O2 photolysis. The most efficient combination of variables was [H2O2] = 255 mg L-1 and [TA] = 25 mg L-1, resulting in 100% TA degradation and 98.87% H2O2 photolysis by 90 min of treatment. Additionally, the second-order kinetic constant of the reaction between TA and HO● was determined using a competitive kinetic model, employing 2,4-dichlorophenoxyacetic acid (2,4D) as the reference compound. The kinetic constant was 1.9 × 1010 M-1 s-1 in alkaline medium. TA degradation by direct photolysis generated quinone imines as by-products, responsible for the formation of a dark red "internal filter" that increased the value of acute toxicity to Artemia salina. The UV-ABC/H2O2 process did not promote formation of quinone imines by 90 min of treatment and therefore did not increase acute toxicity values. Several by-products generated during TA degradation were identified and possible degradation pathways for the UV-ABC and UV-ABC/H2O2 processes were proposed.

The transcription factor p8 regulates autophagy during diapause embryo formation in Artemia parthenogenetica.

Autophagy is an essential homeostatic process by which cytoplasmic components, including macromolecules and organelles, are degraded by lysosome. Increasing evidence suggests that phosphorylated AMP-activated protein kinase (p-AMPK) and target of rapamycin (TOR) play key roles in the regulation of autophagy. However, the regulation of autophagy in quiescent cells remains unclear, despite the fact that autophagy is known to be critical for normal development, regeneration, and degenerative diseases. Here, crustacean Artemia parthenogenetica was used as a model system because they produced and released encysted embryos that enter a state of obligate dormancy in cell quiescence to withstand various environmental threats. We observed that autophagy was increased before diapause stage but dropped to extremely low level in diapause cysts in Artemia. Western blot analyses indicated that the regulation of autophagy was AMPK/TOR independent during diapause embryo formation. Importantly, the level of p8 (Ar-p8), a stress-inducible transcription cofactor, was elevated at the stage just before diapause and was absent in encysted embryos, indicating that Ar-p8 may regulate autophagy. The results of Ar-p8 knockdown revealed that Ar-p8 regulated autophagy during diapause formation in Artemia. Moreover, we observed that activating transcription factors 4 and 6 (ATF4 and ATF6) responded to Ar-p8-regulated autophagy, indicating that autophagy targeted endoplasmic reticulum (ER) during diapause formation in Artemia. Additionally, AMPK/TOR-independent autophagy was validated in human gastric cancer MKN45 cells overexpressing Ar-p8. The findings presented here may provide insights into the role of p8 in regulating autophagy in quiescent cells.

Gamma Radiation Reduced Toxicity of Azoxystrobin Tested on Artemia franciscana.

Fungicide azoxystrobin toxicity was monitored by means of a 96-h biotest with Artemia franciscana nauplius stages after exposure to solutions with concentrations of 0.2, 0.4, 0.6 and 0.8 mg L(-1) irradiated with (60)Co gamma radiation with doses of 1, 2.5, 5 and 10 kGy. The effects of ionization radiation on azoxystrobin toxicity were mainly manifested by a statistically significant reduction of lethality after 72- and 96-h exposure. A maximum reduction of lethality of 72 % was achieved using doses of 1-5 kGy for an azoxystrobin initial concentration of 0.4 mg L(-1) and after 72 h of exposure. At a 96-h exposure, a difference of lethal effects reached up to 70 % for a dose of 10 kGy. The observed effect of gamma ionizing radiation on azoxystrobin toxicity suggest that this approach can be applied as an alternative for a reduction of azoxystrobin residua in food.

Low frequency ultrasound and UV-C for elimination of pathogens in recirculating aquaculture systems.

Low frequency ultrasound (LFUS) was evaluated as a novel disinfection technique within recirculating aquaculture systems both individually and combined with UV-C. Dose-dependent inactivation rates were determined for the total viable counts and model organisms representing different taxa of common fish parasites: the ciliate Paramecium sp., second larval stage (L2) of the nematode Anguillicola crassus and metanauplii of Artemia sp. Application of LFUS up to 19 kJ/L did not reduce the number of colony forming units (CFU), whilst UV-C irradiation was highly effective. Pre-treatment with LFUS reduced the mean size of suspended solids in aquaculture water and thus increased the germicidal effect of UV-C by up to 0.6 log units. LFUS was effective against the eukaryotic organisms, and the dose-dependent inactivation could be well described by functions of an exponential decay. However, the efficiency of LFUS differed greatly between species. A LFUS dose of 1.9 kJ/L (consumed energy) was sufficient to inactivate Artemia by 99%, but a ten times higher dose was necessary to inactivate 95% and 81% of Paramecium and Anguillicola larvae, respectively. In clear water, the energetic efficiency of UV-C (emitted by a low pressure lamp) against Paramecium and Anguillicola larvae was higher compared to LFUS, but LFUS was more efficient against Artemia. However, the efficiency of LFUS against ciliates or nematode larvae would be similar or even higher than UV-C in highly turbid water or if less efficient medium pressure lamps are used. This study shows that LFUS can be applied safely at energy densities that are effective against a wide range of parasites like ciliates, nematodes and crustaceans. The combination of LFUS and UV-C could provide an appropriate water treatment with regards to all relevant pathogens in recirculating aquaculture systems.

Effects of salinity and ultraviolet radiation on the bioaccumulation of mycosporine-like amino acids in Artemia from Lake Urmia (Iran).

We investigated the effects of salinity and artificial UV radiation on the accumulation of mycosporine-like amino acids (MAAs) in sexual and parthenogenetic Artemia from Lake Urmia. The nauplii hatched from the cysts were cultured until adulthood under two salinities (150 and 250 g L(-1) ) and two light treatments (PAR and PAR+UVR) in the laboratory. Finally, the Artemia were analyzed for their concentration of MAAs. In most of the cases, the higher salinity level applied was found to increase the MAA concentrations in both Artemia populations significantly. The acquisition efficiency of MAAs in both Artemia populations increased under exposure to UVR-supplemented photosynthetically active radiation (PAR) compared to those raised under PAR, except for Porphyra-334. It was observed that combination of UV radiation and elevated salinity significantly increased the bioaccumulation of MAAs. Thus, the presence of these compounds in these populations of Artemia may increase their adaptability for living in high-UV and high-salinity conditions prevailing in Lake Urmia. Higher concentrations of MAAs in the parthenogenetic population of Artemia could be probably attributed to its mono sex nature and higher adaptation capacities to extreme environmental conditions.

Selected morphological changes in Artemia franciscana after ionizing radiation exposure.

Nauplii of Artemia franciscana were irradiated by the doses of 0.25, 0.5, 1.0, and 2.5 kGy (60)Co. Dimensions of the body length, body width, intestine width, intestine epithelium width, and intestine lumen width, as well as the mutual ratios of dimensions were determined in 126 specimens. Ratios of the body length/body width (3.98, 3.60, 3.59, and 3.45 vs. 4.13 of control group), and ratios of the intestine epithelium width/intestine lumen width (0.64, 0.52, 0.51, and 0.45 vs. 0.85 of control group), according to the doses, were the most important parameters of evaluation of dependence of morphological changes on radiation doses.

Effects of a new photoactivatable cationic porphyrin on ciliated protozoa and branchiopod crustaceans, potential components of freshwater ecosystems polluted by pathogenic agents and their vectors.

The increasing use of photosensitized processes for disinfection of microbiologically polluted waters requires a precise definition of the factors controlling the degree of photosensitivity in target and non-target organisms. In this regard, tests with protozoa and invertebrates which have a natural habitat in such waters may be used as first screening methods for the assessment of possible hazards for the ecosystem. A new cationic porphyrin, namely meso-tri(N-methyl-pyridyl)mono(N-dodecyl-pyridyl)porphine (C12), is tested in this work on the protozoan Ciliophora Colpoda inflata and Tetrahymena thermophila and the Crustacea Branchiopoda Artemia franciscana and Daphnia magna. The protocol involved 1 h incubation with porphyrin doses in the 0.1-10.0 µM range and subsequent irradiation with visible light at a fluence rate of 10 mW cm(-2). The results indicate that C12 porphyrin has a significant affinity for C. inflata and T. thermophila; this is also shown by fluorescence microscopic analyses. C. inflata cysts were resistant to the phototreatment up to a porphyrin dose of 0.6 µM. The effects of C12 on cysts have been evaluated at 3 and 24 h after the end of the phototreatment; a delay in the excystment process was observed. T. thermophila was fairly resistant to the phototreatment with C12 porphyrin. The data obtained with the two crustaceans indicated that the effects of dark- and photo-treatment with C12 need to be closely examined for every organism. A. franciscana is more resistant, probably owing to its ability to adapt to extreme conditions, while the high level of photosensitivity displayed by Daphnia magna represents a potential drawback, as this organism is often selected as a reference standard for assessing the environmental safety. Thus, while C12 photosensitisation can represent a useful tool for inducing a microbicidal or larvicidal action on polluted waters, the irradiation protocols must be carefully tailored to the nature of the specific water basin, and in particular to its biotic characteristics.

Effects of metal and the phytyl chain on chlorophyll derivatives: physicochemical evaluation for photodynamic inactivation of microorganisms.

Chlorophyll compounds and their derivatives containing metal or phytyl chain can be used as photosensitizer in photodynamic inactivation of microorganisms (PDI). So, the physicochemical properties and antimicrobial effect of chlorophyll derivatives were investigated: Mg-chlorophyll (Mg-Chl), Zn-chlorophyll (Zn-Chl), Zn-chlorophyllide (Zn-Chlde), Cu-chlorophyll (Cu-Chl), pheophytin (Pheo) and pheophorbide (Pheid). The photobleaching experiments showed photostability according to Cu-Chl > Pheo ∼ Pheid ≫ Zn-Chl ∼ Zn-Chlde > Mg-Chl. This order was discussed in terms of metal and the phytyl chain presences. Pheid and Zn-Chl in aqueous Tween 80 solution exhibited highest singlet oxygen yield compared with the other derivatives. Chlorophyll derivatives (CD) with phytyl chain was limited by the self-aggregation phenomenon at high concentrations, even in micellar systems (Tween 80 and P-123). The antimicrobial effect of CD derivatives was investigated against Staphylococcus aureus, Escherichia coli, Candida albicans and Artemia salina. Pheid showed the best results against all organisms tested, Zn-Chlde was an excellent bactericide in the dark and Cu-Chl had no PDI effect. No correlation with CD uptake by microorganisms and darkness cytotoxicity was found. The physicochemical properties allied to bioassays results indicate that Mg-Chl, Pheo, Zn-Chl and Pheid are good candidates for PDI.

Design and implementation of a continuous microwave heating system for ballast water treatment.

A continuous microwave system to treat ballast water inoculated with different invasive species was designed and installed atthe Louisiana State University Agricultural Center. The effectiveness of the system to deliver the required heating loads to inactivate the organisms present was studied. The targeted organisms were microalgae (Nannochloropsis oculata), zooplankton at two different growth stages (newly hatched brine shrimp-Artemia nauplii and adult Artemia), and oyster larvae (Crassosstrea virginica). The system was tested at two different flow rates (1 and 2 liters per min) and power levels (2.5 and 4.5 kW). Temperature profiles indicate that, depending on the species present and the growth stage, the maximum temperature increase will vary from 11.8 to 64.9 degrees C. The continuous microwave heating system delivered uniform and near-instantaneous heating at the outlet proving its effectiveness. The power absorbed and power efficiency varied for the species present. More than 80% power utilization efficiency was obtained at all flow rate and microwave power combinations for microalgae, Artemia nauplii and adults. Test results indicated that microwave treatment can be an effective tool for ballast water treatment, and current high treatment costs notwithstanding, this technique can be added as supplemental technology to the palette of existing treatment methods.

An analysis of dielectric properties of synthetic ballast water at frequencies ranging from 300 to 3000 MHz.

Ballast water presents an important vector for introduction of aquatic invasive species in the coastal waters around the world. Currently there are no established technologies proven to completely eliminate this problem due to the particularities of the ballasting and de-ballasting operations (extremely large volumes of water, efficiency at destroying macro and micro organisms, environmental issues associated with chemical treatments). Continuous microwave heating presents a potential solution to this problem, but the design of suitable applicators depends on the dielectric properties of the ballast water to be processed. The study presented in this paper is focused on the dielectric properties (dielectric constant--epsilon'; dielectric loss--epsilon") of synthetic ballast water inoculated with four organisms at seven different temperatures in the frequency range of 300 to 3000 MHz. The dielectric properties of the mixtures were determined using a network analyzer and a dielectric probe kit using the open-ended coaxial probe method. Numerical analysis was performed on data collected across all frequencies involved with an emphasis placed on F.C.C. allotted frequencies of 433, 915 and 2450 MHz. The dielectric constant was relatively independent of frequency and the organism used, but it showed a remarkable decrease with temperature. The dielectric loss showed an extreme decrease with increasing frequency, marked differences between the different organisms and between different growth stages of the same organism, and a large relatively linear increase with increasing temperature.

A continuous microwave system for prevention of invasive species during de-ballasting operation--death kinetics.

A continuous microwave heating system was tested for its effectiveness at removing potentially invasive organisms during deballasting operations. Four different organisms, namely Nannochloropsis oculata (microalgae), Artemia nauplii, Artemia adults and Crassosstrea virginica (oyster larvae) normally found in ballast water were investigated in a controlled study to quantify their survival after continuous microwave heating of synthetic ballast water. The experiments were performed in the microwave system using a 2 x 2 factorial design with power (2.5 and 4.5 kW) and flow rate (1.0 and 2.0 lpm) and the organisms subsequently subjected to different holding times. The control treatment was performed in a water bath using the same temperatures and holding times as in the case of the microwave treatment. Overall, the results obtained indicated that the microwave system was more effective in eliminating the organisms when compared with the control treatment. In most cases there were no survivors present after the microwave treatment at holding times above 100 s, and temperatures as low as 50 degrees C particularly for oyster larvae and Artemia adults. The results are promising, indicating that this technology has the potential to be an effective tool in controlling/preventing the introduction of invasive species into native environments.

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.

Estimate of the effects of ultraviolet radiation on the mortality of Artemia franciscana in naupliar and adult stages.

The impact of different doses of artificial ultraviolet (UV) radiation on the growth stages of a marine zooplankton was investigated using laboratory microcosms. Mortality percentages of naupliar and adult samples of Artemia franciscana were recorded in relation to different UV doses (single exposure: 75, 150, 300, 600, 1,200, 2,400, 3,900, 7,800 J m(-2)) at specific observation times after exposure (24, 48, 72, 96 and 120 h). The relationship between mortality percentage and UV dose showed significant differences in relation to the zooplankton growth stage. The elevated susceptibility of the naupliar samples to UV radiation is described through a mortality model based on a logistic equation. The data analysis shows that the slope of mortality versus dose remains the same for the two growth stages while the lethal dose in the naupliar stage was 3.3 smaller than that determined for the adult stage. The slope of the UV mortality rate versus post-incubation time was found to be significantly different (P<0.05) at low UV doses for the two life stages examined, i.e. naupliar and adult. The lower value of LD(50) in naupliar stages compared to that for adults confirms that in the early growth stage this marine zooplankton is more susceptible to UV radiation.

Neutron flow exposure as a test for survival of Artemia salina spores.

Live and heat-inactivated Artemia salina spores (samples with the same mass and filling density) were exposed to a flow of thermal neutrons from a (252)Cf radioactive source at an equivalent dose power of about 1 microSv/h. Irradiation led to a 4-fold acceleration of nauplius development and to modification of the element profiles of live spores. The difference between absorption/diffusion of thermal neutrons by live and dead spores was revealed.

Alkoxypsoralens, novel nonpeptide blockers of Shaker-type K+ channels: synthesis and photoreactivity.

A series of psoralens and structurally related 5,7-disubstituted coumarins was synthesized and investigated for their K+ channel blocking activity as well as for their phototoxicity to Artemia salina and their ability to generate singlet oxygen and to photomodify DNA. After screening the compounds on Ranvier nodes of the toad Xenopus laevis, the affinities of the most promising compounds, which proved to be psoralens bearing alkoxy substituents in the 5-position or alkoxymethyl substituents in the neighboring 4- or 4'-position, to a number of homomeric K+ channels were characterized. All compounds exhibited the highest affinity to Kv1.2. 5,8-Diethoxypsoralen (10d) was found to be an equally potent inhibitor of Kv1.2 and Kv1.3, while lacking the phototoxicity normally inherent in psoralens. The reported compounds represent a novel series of nonpeptide blockers of Shaker-type K+ channels that could be further developed into selective inhibitors of Kv1.2 or Kv1. 3.

Results of space experiments.

Life science research in space was started in Europe with the first Biostack experiment flown onboard Apollo 16 in 1972. Biostack was designed to investigate the biological effects of single heavy ions of cosmic radiation. Among several undertakings towards this goal, the Biostack achieved the highest precision in the determination of the spatial correlation of the observed biological response of single test organisms to the passage of single heavy ions, which is the mandatory requirement. It also provided information on the influence of additional spaceflight factors, such as microgravity, on radiation effects and measurements of the spectrum of charge and energy of the cosmic radiation. The experiment was performed as an international cooperation effort. This report gives a summary of the biological data accumulated in this and the follow-on experiments of the Biostack program.

Influence of a long duration exposure, 69 months, to the space flight factors in Artemia cysts, tobacco and rice seeds.

Three french laboratories have participated in the Free Flyer Biostack experiment. Artemia cysts, tobacco seeds and rice caryopsis and embryos were used. Biological objects in monolayers were dead. In opposite, a large fraction of samples used in bulk survived. A stimulatory effect occurred in the first steps of development in Artemia cysts. In fact, the larval survival was unchanged or slightly reduced. In tobacco a drastic decrease in germination and survival rate was observed. Space flight did not induce genetic changes. In rice, results depend on the variety which was investigated; the growth rate stimulation in flight samples is discussed with respect to controls.

Recent approaches in the analysis of weightlessness effects on arthropod development.

The concept of Biological Development refers to the extremely complex process by which every biological organism reproduces starting from a huge single cell, the fertilized egg. It includes all aspects of cellular and intercellular structure and function. In spite of many recent advances, especially at the molecular and genetical level, we are still far from fully understanding the details and mechanisms at work in developmental systems. It is even unclear what physical mechanisms are used by the different molecular components resulting in the emergence of these higher levels of organization. Newman and Comper, have extensively discussed the "generic" physical forces potentially involved in pattern formation, arguing that among others, gravitational effects could be involved in the production of cytoplasmic, tissue and extracellular matrix components rearrangements playing a role in morphogenesis. Although plagued with the problem of being a very weak force, specially at the tiny dimensions of cells, gravity is one of the "generic" physical forces that have been continuously operating on biological organisms during evolution. Few scientists would argue against the idea that at least in the early times of evolution, gravity could have been involved in shaping the spatial inhomogeneities behind the initial phases of development.

The effect of krypton ions on Artemia cysts.

Dry eggs of Artemia salina were exposed to 80 KV-X-rays and Kr-Ions (energy: 11.75 MeV/u; LET: 3500 KeV/microns). Emergence, hatching and survival were determined. The results demonstrate a very high resistance both to sparsely and ionizing radiation.