Search results for: biological synthesis

Authors: Sofia Lazareva, Artem Smolentsev

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Fluorescent photochromic materials collect strong interest due to their possible application in organic photonics such as optical logic systems, optical memory, visualizing sensors, as well as characterization of polymers and biological systems. In photochromic fluorescence switching systems the emission of fluorophore is modulated between ‘on’ and ‘off’ via the photoisomerization of photochromic moieties resulting in effective resonance energy transfer (FRET). In current work, we have studied both photochromic and fluorescent properties of several diarylethenes. It was found that coloured forms of these compounds are not fluorescent because of the efficient intramolecular energy transfer. Spectral and photochromic parameters of investigated substances have been measured in five solvents having different polarity. Quantum yields of photochromic transformation A↔B ΦA→B and ΦB→A as well as B isomer extinction coefficients were determined by kinetic method. It was found that the photocyclization reaction quantum yield of all compounds decreases with the increase of solvent polarity. In addition, the solvent polarity is revealed to affect fluorescence significantly. Increasing of the solvent dielectric constant was found to result in a strong shift of emission band position from 450 nm (nhexane) to 550 nm (DMSO and ethanol) for all three compounds. Moreover, the emission intensive in polar solvents becomes weak and hardly detectable in n-hexane. The only one exception in the described dependence is abnormally low fluorescence quantum yield in ethanol presumably caused by the loss of electron-donating properties of nitrogen atom due to the protonation. An effect of the protonation was also confirmed by the addition of concentrated HCl in solution resulting in a complete disappearance of the fluorescent band. Excited state dynamics were investigated by ultrafast optical spectroscopy methods. Kinetic curves of excited states absorption and fluorescence decays were measured. Lifetimes of transient states were calculated from the data measured. The mechanism of ring opening reaction was found to be polarity dependent. Comparative analysis of kinetics measured in acetonitrile and hexane reveals differences in relaxation dynamics after the laser pulse. The most important fact is the presence of two decay processes in acetonitrile, whereas only one is present in hexane. This fact supports an assumption made on the basis of steady-state preliminary experiments that in polar solvents occur stabilization of TICT state. Thus, results achieved prove the hypothesis of two channel mechanism of energy relaxation of compounds studied.

Keywords: diarylethenes, fluorescence switching, FRET, photochromism, TICT state

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Authors: A. Kesraoui, M. Seffen

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Applications of adsorption onto activated carbon for water treatment are well known. The process has been demonstrated to be widely effective for removing dissolved organic substances from wastewaters, but this treatment has a major drawback is the high operating cost. The main goal of our research work is to improve the retention capacity of Tunisian biomass for the depollution of industrial wastewater and retention of pollutants considered toxic. The biosorption process is based on the retention of molecules and ions onto a solid surface composed of biological materials. The evaluation of the potential use of these materials is important to propose as an alternative to the adsorption process generally expensive, used to remove organic compounds. Indeed, these materials are very abundant in nature and are low cost. Certainly, the biosorption process is effective to remove the pollutants, but it presents a slow kinetics. The improvement of the biosorption rates is a challenge to make this process competitive with respect to oxidation and adsorption onto lignocellulosic fibers. In this context, the alternating current appears as a new alternative, original and a very interesting phenomenon in the acceleration of chemical reactions. Our main goal is to increase the retention acceleration of dyes (indigo carmine, methylene blue) and phenol by using a new alternative: alternating current. The adsorption experiments have been performed in a batch reactor by adding some of the adsorbents in 150 mL of pollutants solution with the desired concentration and pH. The electrical part of the mounting comprises a current source which delivers an alternating current voltage of 2 to 15 V. It is connected to a voltmeter that allows us to read the voltage. In a 150 mL capacity cell, we plunged two zinc electrodes and the distance between two Zinc electrodes has been 4 cm. Thanks to alternating current, we have succeeded to improve the performance of activated carbon by increasing the speed of the indigo carmine adsorption process and reducing the treatment time. On the other hand, we have studied the influence of the alternating current on the biosorption rate of methylene blue onto Luffa cylindrica fibers and the hybrid material (Luffa cylindrica-ZnO). The results showed that the alternating current accelerated the biosorption rate of methylene blue onto the Luffa cylindrica and the Luffa cylindrica-ZnO hybrid material and increased the adsorbed amount of methylene blue on both adsorbents. In order to improve the removal of phenol, we performed the coupling between the alternating current and the biosorption onto two adsorbents: Luffa cylindrica and the hybrid material (Luffa cylindrica-ZnO). In fact, the alternating current has succeeded to improve the performance of adsorbents by increasing the speed of the adsorption process and the adsorption capacity and reduce the processing time.

Keywords: adsorption, alternating current, dyes, modeling

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Authors: Muhammad Naeem Chaudhry, Fahim Nawaz, Rana Nauman Shabbir

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New strategies aimed at increasing the resilience of crop plants to the negative effects of climate change represent important research priorities of plant scientists. The use of soil microorganisms to alleviate abiotic stresses like drought has gained particular importance in recent past. A field experiment was planned to investigate the effect of phosphorous solubilizing bacteria on yield and seed quality of Camelina (Camelina sativa L.) under water deficit conditions. The study was conducted at Agronomic Research Farm, University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur, during 4th week of November, 2013. The available seeds of Camelina sativa were inoculated with two bacterial strains (pseudomonas and Bacillus spp.) and grown under various water stress levels i.e. D0, (four irrigations), D3 (three irrigation), D2 (two irrigations), and D1 (one irrigation). The results revealed that drought stress significantly reduced the plant growth and yield, consequently reducing protein contents and oil concentration in camelina. The exposure to drought stress decreased plant height (16%), plant population (27%), number of fertile branches (41-59%), number of pods per plant (35%) and seed per pod (33%). Drought stress also exerted a negative impact on yield characteristics by reducing the 1000-seed weight (65%), final seed yield (52%), biological yield (22%) and harvest index (39%) of camelina. However, the inoculation of seeds with Pseudomonas and Bacillus spp. promoted the plant growth characterized by increased plant height and enhanced plant population. It was noted that inoculation of seeds with Pseudomonas resulted in the maximum plant population (113.4 cm), primary branches (19 plant-1), and number of pods (664 plant-1), whereas Bacillus inoculation resulted in maximum plant height (113.4 cm), seeds per pod (15.9), 1000-seed weight (1.85 g), and seed yield (3378.8 kg ha-1). Moreover, the inoculation with Bacillus also significantly improved the quality attributes of camelina and gave 3.5% and 2.1% higher oil contents than Pseudomonas and control (no-inoculation), respectively. Similarly, the same strain also resulted in maximum protein contents (33.3%). Our results confirmed the hypothesis that inoculation of seeds with phosphorous solubilizing bacterial strains is an effective, viable and environment-friendly approach to improve yield and quality of camelina under water deficit conditions. However, further studies are suggested to investigate the physiological and molecular processes, stimulated by bacterial strains, for increasing drought tolerance in food crops.

Keywords: Camelina, drought stress, phosphate solubilizing bacteria, seed quality

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Authors: Abdolamir Allameh, Shahnaz Esmaeili, Mina Allameh, Safoura Khajeniazi

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Stem cells with therapeutic applications can be isolated from human placenta/umblical cord blood (UCB) as well as the cord tissue (UC). Stem cells in culture are vulnerable to oxidative stress, particularly when subjected to differentiation process. The aim of this study was to examine the chnages in the rate of oxidation that occurs to cellular macromolecules during hepatic differentiation of mononuclear cells (MSCs). In addition, the impact of the hepatic differentiation process of MSC on cellular and biological activity of the cells will be undertaken. For this purpose, first mononuclear cells (MNCs) were isolated from human UCB which was obtained from a healthy full-term infant. The cells were cultured at a density of 3×10⁵ cells/cm² in DMEM- low-glucose culture media supplemented with 20% FBS, 2 mM L-glutamine, 100 μg/ml streptomycin and 100 U/ml penicillin. Cell cultures were then incubated at 37°C in a humidified 5% CO₂ incubator. After removing non-adherent cells by replacing culture medium, fibroblast-like adherent cells were resuspended in 0.25% trypsin-EDTA and plated in 25 cm² flasks (1×10⁴/ml). Characterization of the MSCs was routinely done by observing their morphology and growth curve. MSCs were subjected to a 2-step hepatocyte differentiation protocol in presence of hepatocyte growth factor (HGF), dexamethazone (DEX) and oncostatin M (OSM). The hepatocyte-like cells derived from MSCs were checked every week for 3 weeks for changes in lipid peroxidation, protein carbonyl formation and DNA oxidation i.e., 8-hydroxy-2'-deoxyguanosine (8-OH-dG) assay. During the 3-week differentiation process of MSCs to hepatocyte-like cells we found that expression liver-specific markers such as albumin, was associated with increased levels of lipid peroxidation and protein carbonyl formation. Whereas, undifferentiated MSCs has relatively low levels of lipid peroxidation products. There was a significant increase ( p < 0.05) in lipid peroxidation products in hepatocytes on days 7, 14, and 21 of differentiation. Likewise, the level of protein carbonyls in the cells was elevated during the differentiation. The level of protein carbonyls measured in hepatocyte-like cells obtained 3 weeks after differentiation induction was estimated to be ~6 fold higher compared to cells recovered on day 7 of differentiation. On the contrary, there was a small but significant decrease in DNA damage marker (8-OH-dG) in hepatocytes recovered 3 weeks after differentiation onset. The level of 8-OHdG which was in consistent with formation of reactive oxygen species (ROS). In conclusion, this data suggest that despite the elevation in oxidation of lipid and protein molecules during hepatocyte development, the cells were normal in terms of DNA integrity, morphology, and biologically activity.

Keywords: adult stem cells, DNA integrity, free radicals, hepatic differentiation

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Authors: E. Rommes, Els Toonen, Rahil Roodsaz, Suborna Camellia, Farhana Alam, Saad Khan, Jhalok Ranjon Talukder, Tanveer Hassan, Syeda Farjana Ahmed, Sabina Faiz Rashid

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In Bangladesh, a country in which adults (both parents and teachers) find it particularly hard to speak with youth about sexuality, adolescents seem to struggle with various insecurities about their sexual feelings, thoughts, behavior and physical characteristics. On the basis of a large number of interviews and focus groups with rural and urban Bangla adolescent girls and boys of lower and middle class as part of the large-scale three-year project ‘Breaking the Shame’, we have identified ten sexual themes or ‘myths’ that youth struggle with most. These encompass amongst others beliefs and insecurities on masturbation, discharge, same-sex behavior and feelings, the effects of watching porn and gender norms. We argue that the Internet is a particularly suitable medium to ‘debunk’ those myths, as youth can consult it anonymously and privately and so avoid social shame. Moreover, amongst the myths, we have identified two kinds which may need different debunking techniques. One kind of myth concerns scientifically uncontested, generally biological related information, such as the effects of having sex with a pregnant woman, questions on the effects of a penile or vaginal discharge or questions on the effects of masturbation. The second kind of myths concerns more diverse information sources and deals with e.g. religious or culturally specific norms, such as on the meaning and existence of homosexuality or gender appropriate norms of behavior in Bangladesh. For addressing both kinds of myths, expert information including a wealth of references to information resources needs to be provided, which the Internet is very suitable for. For the second kind of myths, adolescents also need to learn how to deal with sometimes conflicting norms and information sources, and they need to develop and reflect on their own opinions as part of their identity formation. On the basis of a literature review, we thus distinguish general information needs from identity formation needs, which includes the need to be able to relate information and opinions to one’s own opinions and situation. Hence, we argue that youth not only need abstract expert information to be able to debunk sexual myths, but also the option to discuss this information with other adolescents and compare their own situation and opinions with other peers, who in that way serve as ‘warm experts’ for each other. In this paper, we will describe the outcomes of our qualitative study above. In addition, we will present our findings of an intervention by presenting youth with general, uncontested information on the Internet with additional peer discussion options to compare the debunking effects on different kinds of myths.

Keywords: peer discussion, intervention, sexual myths, shame

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Authors: N. Selvi Gunel, L. M. Oktay, H. Memmedov, B. Durmaz, H. Kalkan Yildirim, E. Yildirim Sozmen

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Object: Propolis which consists of compounds that are accepted as antioxidant, antimicrobial, antiseptic, antibacterial, anti-inflammatory, anti-mutagenic, immune-modulator and cytotoxic, is frequently used in current therapeutic applications. However, some of them result in allergic side effects, causing consumption to be restricted. Previously our group has succeeded in producing a new biotechnological product which was less allergenic. In this study, we purpose to optimize production conditions of this biologically-transformed propolis and determine the cytotoxic effects of obtained new products on colon cancer cell line (HCT-116). Method: Firstly, solid propolis samples were dissolved in water after weighing, grinding and sizing (sieve-35mesh) and applied 40 kHz/10 min ultrasonication. Samples were prepared according to inoculation with Lactobacillus plantarum in two different proportions (2.5% and 3.5%). Chromatographic analyzes of propolis were performed by UPLC-MS/MS (Waters, Milford, MA) system. Results were analysed by UPLC-MS/MS system MassLynx™ 4.1 software. HCT-116 cells were treated with propolis examples at 25-1000 µg/ml concentrations and cytotoxicity were measured by using WST-8 assay at 24, 48, and 72 hours. Samples with biological transformation were compared with the non-transformed control group samples. Our experiment groups were formed as follows: untreated (group 1), propolis dissolved in water ultrasonicated at 40 kHz/10 min (group 2), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 2.5% L. plantarum L1 strain (group 3), propolis dissolved in water ultrasonicated at 40 kHz/10 min and inoculated 3.5% L. plantarum L3 strain (group 4). Obtained data were calculated with Graphpad Software V5 and analyzed by two-way ANOVA test followed by Bonferroni test. Result: As a result of our study, the cytotoxic effect of propolis samples on HCT-116 cells was evaluated. There was a 7.21 fold increase in group 3 compared to group 2 in the concentration of 1000 µg/ml, and it was a 6.66 fold increase in group 3 compared to group 1 at the end of 24 hours. At the end of 48 hours, in the concentration of 500 µg/ml, it was determined 4.7 fold increase in group 4 compared to group 3. At the same time, in the concentration of 750 µg/ml it was determined 2.01 fold increase in group 4 compared to group 3 and in the same concentration, it was determined 3.1 fold increase in group 4 compared to group 2. Also, at the 72 hours, in the concentration of 750 µg/ml, it was determined 2.42 fold increase in group 3 according to group 2 and in the same time, in the concentration of 1000 µg/ml, it was determined 2.13 fold increase in group 4 according to group 2. According to cytotoxicity results, the group which were ultrasonicated at 40 kHz/10min and inoculated 3.5% L. plantarum L3-strain had a higher cytotoxic effect. Conclusion: It is known that bioavailability of propolis is halved in six months. The data obtained from our results indicated that biologically-transformed propolis had more cytotoxic effect than non-transformed group on colon cancer cells. Consequently, we suggested that L. plantarum-transformation provides both reduction of allergenicity and extension of bioavailability period by enhancing healthful polyphenols.

Keywords: bio-transformation, propolis, colon cancer, cytotoxicity

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Authors: Mmatladi D. Mnguni, Mustapha Mohammed, George Y. Mahama, Alhassan L. Abdulai, Felix D. Dakora

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Industrial-based nitrogen (N) fertilizers are justifiably credited for the current state of food production across the globe, but their continued use is not sustainable and has an adverse effect on the environment. The search for greener and sustainable technologies has led to an increase in exploiting biological systems such as legumes and organic amendments for plant growth promotion in cropping systems. Although the benefits of legume rotation with cereal crops have been documented, the full benefits of soybean-sorghum rotation systems have not been properly evaluated in Africa. This study explored the benefits of soybean-sorghum rotation through assessing N₂ fixation and water-use efficiency of soybean in rotation with sorghum with and without organic and inorganic amendments. The field trials were conducted from 2017 to 2020. Sorghum was grown on plots previously cultivated to soybean and vice versa. The succeeding sorghum crop received fertilizer amendments [organic fertilizer (5 tons/ha as poultry litter, OF); inorganic fertilizer (80N-60P-60K) IF; organic + inorganic fertilizer (OF+IF); half organic + inorganic fertilizer (HIF+OF); organic + half inorganic fertilizer (OF+HIF); half organic + half inorganic (HOF+HIF) and control] and was arranged in a randomized complete block design. The soybean crop succeeding fertilized sorghum received a blanket application of triple superphosphate at 26 kg P ha⁻¹. Nitrogen fixation and water-use efficiency were respectively assessed at the flowering stage using the ¹⁵N and ¹³C natural abundance techniques. The results showed that the shoot dry matter of soybean plants supplied with HOF+HIF was much higher (43.20 g plant-1), followed by OF+HIF (36.45 g plant⁻¹), and HOF+IF (33.50 g plant⁻¹). Shoot N concentration ranged from 1.60 to 1.66%, and total N content from 339 to 691 mg N plant⁻¹. The δ¹⁵N values of soybean shoots ranged from -1.17‰ to -0.64‰, with plants growing on plots previously treated to HOF+HIF exhibiting much higher δ¹⁵N values, and hence lower percent N derived from N₂ fixation (%Ndfa). Shoot %Ndfa values varied from 70 to 82%. The high %Ndfa values obtained in this study suggest that the previous year’s organic and inorganic fertilizer amendments to sorghum did not inhibit N₂ fixation in the following soybean crop. The amount of N-fixed by soybean ranged from 106 to 197 kg N ha⁻¹. The treatments showed marked variations in carbon (C) content, with HOF+HIF treatment recording the highest C content. Although water-use efficiency varied from -29.32‰ to -27.85‰, shoot water-use efficiency, C concentration, and C:N ratio were not altered by previous fertilizer application to sorghum. This study provides strong evidence that previous HOF+HIF sorghum residues can enhance N nutrition and water-use efficiency in nodulated soybean.

Keywords: ¹³C and ¹⁵N natural abundance, N-fixed, organic and inorganic fertilizer amendments, shoot %Ndfa

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Authors: V. Stejskal, K. Lundova, R. Sebesta, T. Vanina, S. Roje

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Pure strain of Arctic char (AC) Salvelinus alpinus and hybrid (HB) Salvelinus alpinus vs. Salvelinus fontinalis Mitchill belong to fish, which with great potential for culture in recirculating aquaculture systems (RAS). Aquaculture of these fish currently use flow-through systems (FTS), especially in Nordic countries such as Iceland (biggest producer), Norway, Sweden, and Canada. Four different water saturation regimes included normoxia (NOR), permanent hyperoxia (HYP), intermittent hyperoxia (HYP ± ) and regimes where one day of normoxia was followed by one day of hyperoxia (HYP1/1) were tested during 63 days of experiment in both species in two parallel experiments. Fish were reared in two identical RAS system consisted of 24 plastic round tanks (300 L each), drum filter, biological filter with moving beads and submerged biofilter. The temperature was maintained using flow-through cooler during at level of 13.6 ± 0.8 °C. Different water saturation regimes were achieved by mixing of pure oxygen (O₂) with water in three (one for each hyperoxic regime) mixing tower equipped with flowmeter for regulation of gas inflow. The water in groups HYP, HYP1/1 and HYP± was enriched with oxygen up to saturation of 120-130%. In HYP group was this level kept during whole day. In HYP ± group was hyperoxia kept for daylight phase (08:00-20:00) only and during night time was applied normoxia in this group. The oxygen saturation of 80-90% in NOR group was created using intensive aeration in header tank. The fish were fed with commercial feed to slight excess at 2 h intervals within the light phase of the day. Water quality parameters like pH, temperature and level of oxygen was monitoring three times (7 am, 10 am and 6 pm) per day using handy multimeter. Ammonium, nitrite and nitrate were measured in two day interval using spectrophotometry. Initial body weight (BW) was 40.9 ± 8.7 g and 70.6 ± 14.8 in AC and HB group, respectively. Final survival of AC ranged from 96.3 ± 4.6 (HYP) to 100 ± 0.0% in all other groups without significant differences among these groups. Similarly very high survival was reached in trial with HB with levels from 99.2 ± 1.3 (HYP, HYP1/1 and NOR) to 100 ± 0.0% (HYP ± ). HB fish showed best growth performance in NOR group reached final body weight (BW) 180.4 ± 2.3 g. Fish growth under different hyperoxic regimes was significantly reduced and final BW was 164.4 ± 7.6, 162.1 ± 12.2 and 151.7 ± 6.8 g in groups HY1/1, HYP ± and HYP, respectively. AC showed different preference for hyperoxic regimes as there were no significant difference in BW among NOR, HY1/1 and HYP± group with final values of 72.3 ± 11.3, 68.3 ± 8.4 and 77.1 ± 6.1g. Significantly reduced growth (BW 61.8 ± 6.8 g) was observed in HYP group. It is evident from present study that there are differences between pure bred Arctic char and hybrid in relation to hyperoxic regimes. The study was supported by projects 'CENAKVA' (No. CZ.1.05/2.1.00/01.0024), 'CENAKVA II' (No. LO1205 under the NPU I program), NAZV (QJ1510077) and GAJU (No. 060/2016/Z).

Keywords: recirculating aquaculture systems, Salmonidae, hyperoxia, abiotic factors

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Authors: Helen Nelly Naiga, Jane Frances Zalwango, Saudah N. Kizito, Brian Agaba, Brenda N Simbwa, Maria Goretti Zalwango, Richard Migisha, Benon Kwesiga, Daniel Kadobera, Alex Ario Riolexus, Sarah Paige, Julie R. Harris

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Background: Traditional community beliefs and practices can facilitate the spread of Ebola virus during outbreaks. On September 20, 2022, Uganda declared a Sudan Virus Disease (SVD) outbreak after a case was confirmed in Mubende District. During September–November 2022, the outbreak spread to eight additional districts. We investigated the role of community beliefs and practices in the spread of SUDV in Uganda in 2022. Methods: A qualitative study was conducted in Mubende, Kassanda, and Kyegegwa districts in February 2023. We conducted nine focus group discussions (FGDs) and six key informant interviews (KIIs). FGDs included SVD survivors, household members of SVD patients, traditional healers, religious leaders, and community leaders. Key informants included community, political, and religious leaders, traditional healers, and health workers. We asked about community beliefs and practices to understand if and how they contributed to the spread of SUDV. Interviews were recorded, translated, transcribed, and analyzed thematically. Results: Frequently-reported themes included beliefs that the community deaths, later found to be due to SVD, were the result of witchcraft or poisoning. Key informants reported that SVD patients frequently first consulted traditional healers or spiritual leaders before seeking formal healthcare, and noted that traditional healers treated patients with signs and symptoms of SVD without protective measures. Additional themes included religious leaders conducting laying-on-of-hands prayers for SVD patients and symptomatic contacts, SVD patients and their symptomatic contacts hiding in friends’ homes, and exhumation of SVD patients originally buried in safe and dignified burials, to enable traditional burials. Conclusion: Multiple community beliefs and practices likely promoted SVD outbreak spread during the 2022 outbreak in Uganda. Engaging traditional and spiritual healers early during similar outbreaks through risk communication and community engagement efforts could facilitate outbreak control. Targeted community messaging, including clear biological explanations for clusters of deaths and information on the dangers of exhuming bodies of SVD patients, could similarly facilitate improved control in future outbreaks in Uganda.

Keywords: Ebola, Sudan virus, outbreak, beliefs, traditional

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Authors: Swati Tomar, Sunil Kumar Gupta

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Anammox is a novel and promising technology that has changed the traditional concept of biological nitrogen removal. The process facilitates direct oxidation of ammonical nitrogen under anaerobic conditions with nitrite as an electron acceptor without the addition of external carbon sources. The present study investigated the feasibility of anammox hybrid reactor (AHR) combining the dual advantages of suspended and attached growth media for biodegradation of ammonical nitrogen in wastewater. The experimental unit consisted of 4 nos. of 5L capacity AHR inoculated with mixed seed culture containing anoxic and activated sludge (1:1). The process was established by feeding the reactors with synthetic wastewater containing NH4-H and NO2-N in the ratio 1:1 at HRT (hydraulic retention time) of 1 day. The reactors were gradually acclimated to higher ammonium concentration till it attained pseudo steady state removal at a total nitrogen concentration of 1200 mg/l. During this period, the performance of the AHR was monitored at twelve different HRTs varying from 0.25-3.0 d with increasing NLR from 0.4 to 4.8 kg N/m3d. AHR demonstrated significantly higher nitrogen removal (95.1%) at optimal HRT of 1 day. Filter media in AHR contributed an additional 27.2% ammonium removal in addition to 72% reduction in the sludge washout rate. This may be attributed to the functional mechanism of filter media which acts as a mechanical sieve and reduces the sludge washout rate many folds. This enhances the biomass retention capacity of the reactor by 25%, which is the key parameter for successful operation of high rate bioreactors. The effluent nitrate concentration, which is one of the bottlenecks of anammox process was also minimised significantly (42.3-52.3 mg/L). Process kinetics was evaluated using first order and Grau-second order models. The first-order substrate removal rate constant was found as 13.0 d-1. Model validation revealed that Grau second order model was more precise and predicted effluent nitrogen concentration with least error (1.84±10%). A new mathematical model based on mass balance was developed to predict N2 gas in AHR. The mass balance model derived from total nitrogen dictated significantly higher correlation (R2=0.986) and predicted N2 gas with least error of precision (0.12±8.49%). SEM study of biomass indicated the presence of the heterogeneous population of cocci and rod shaped bacteria of average diameter varying from 1.2-1.5 mm. Owing to enhanced NRE coupled with meagre production of effluent nitrate and its ability to retain high biomass, AHR proved to be the most competitive reactor configuration for dealing with nitrogen laden wastewater.

Keywords: anammox, filter media, kinetics, nitrogen removal

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Authors: Antonina A. Shumakova, Sergey A. Khotimchenko

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The relevance of research is associated, on the one hand, with an ever-increasing volume of production and the expansion of the scope of application of engineered nanomaterials (ENMs), and on the other hand, with the lack of sufficient scientific information on the nature of the interactions of nanoparticles (NPs) with components of biogenic and abiogenic origin. In particular, studying the effect of ENMs (TiO2 NPs, SiO2 NPs, Al2O3 NPs, fullerenol) on the toxicometric characteristics of common contaminants such as lead and cadmium is an important hygienic task, given the high probability of their joint presence in food products. Data were obtained characterizing a multidirectional change in the toxicity of model toxicants when they are co-administered with various types of ENMs. One explanation for this fact is the difference in the adsorption capacity of ENMs, which was further studied in in vitro studies. For this, a method was proposed based on in vitro modeling of conditions simulating the environment of the small intestine. It should be noted that the obtained data are in good agreement with the results of in vivo experiments: - with the combined administration of lead and TiO2 NPs, there were no significant changes in the accumulation of lead in rat liver; in other organs (kidneys, spleen, testes and brain), the lead content was lower than in animals of the control group; - studying the combined effect of lead and Al2O3 NPs, a multiple and significant increase in the accumulation of lead in rat liver was observed with an increase in the dose of Al2O3 NPs. For other organs, the introduction of various doses of Al2O3 NPs did not significantly affect the bioaccumulation of lead; - with the combined administration of lead and SiO2 NPs in different doses, there was no increase in lead accumulation in all studied organs. Based on the data obtained, it can be assumed that at least three scenarios of the combined effects of ENMs and chemical contaminants on the body: - ENMs quite firmly bind contaminants in the gastrointestinal tract and such a complex becomes inaccessible (or inaccessible) for absorption; in this case, it can be expected that the toxicity of both ENMs and contaminants will decrease; - the complex formed in the gastrointestinal tract has partial solubility and can penetrate biological membranes and / or physiological barriers of the body; in this case, ENMs can play the role of a kind of conductor for contaminants and, thus, their penetration into the internal environment of the body increases, thereby increasing the toxicity of contaminants; - ENMs and contaminants do not interact with each other in any way, therefore the toxicity of each of them is determined only by its quantity and does not depend on the quantity of another component. Authors hypothesized that the degree of adsorption of various elements on the surface of ENMs may be a unique characteristic of their action, allowing a more accurate understanding of the processes occurring in a living organism.

Keywords: absorption, cadmium, engineered nanomaterials, lead

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Authors: Serap Gedikli, Hakan Çakmak, M. Buğra Güldiken, Duygu Yalnızoğlu

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Air, which is necessary for living things to survive; while it carries some useful substances in it, it can also carry foreign particles of different sizes that may be harmful to the health. All airborne organic substances of biological origin, including bacteria, fungi, fungal spores, viruses, pollen, and their components, are called "bioaerosols". Nowadays, everyone spends most of their time in closed areas such as home, workplace, school, etc. Although it is known that outdoor air pollution affects health, it is not known that indoor air pollution has harmful effects in terms of health. In this study, indoor air microbial load and SARS-CoV-2 virus cleaning efficiency of Froumann brand air cleaners were studied. This work in 300 m³, 600 m³, and 1000 m³ completely closed areas without any air circulation with Froumann N80, N90, and N100 air-cleaning devices. Analyzes were performed for both areas at 60 minutes before and after the device was operated using a particle measuring device (Particles Plus 7302) and an air sampler (Mas-100 ECO). The measurements were taken by placing the test equipment 1.5-2 m away from the air cleaner. At the same time, the efficiency of the HEPA filter was evaluated by taking samples from the air outlet point of the HEPA filter using the air sampling device (Mas-100 ECO) after the device was started. Nutrient agar and malt agar are used as total mesophilic bacteria and total fungi. The number of colony-forming units per m³ (cfu/m³) was calculated by counting colonies in Petri dishes after incubation for 48 hours at 37°C for bacteria and 72 hours at 30°C for fungi. The change in the number of colonies and the decrease in the microbial load was calculated as a percentage value. SARS-CoV-2 activity analysis studies were carried out by İnönü University Microbiology Department in accordance with the World Health Organization regulations. Finally, the HEPA filter in the devices used was taken and kept under a certain temperature and humidity, and the change in the microbial load on it was monitored over a 6-month period. At the end of the studies, a 91%-94% reduction was determined in the total mesophilic bacteria count of Frouman brand N80, N90, and N100 model air cleaners. A decrease of 94%-96% was detected in the total number of yeast/molds. HEPA filter efficiency was evaluated, and at the end of the analysis, 98% of the bacterial load and approximately 100% of yeast/mold load at the HEPA filter air outlet point were decreased. According to the SARS- CoV-2 analysis results, when the device is operating at the medium airflow level 3, it can filter virus-carrying aerosols by 99%. As a result, it was determined that the Froumann model air cleaner was effective in controlling and reducing the microbial load in the indoor air.

Keywords: HEPA filter, indoor air quality, microbial load, SARS-CoV-2

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Authors: Bruno C. Schimming, Leandro L. Martins, PatríCia F. F. Pinheiro, Raquel F. Domeniconi, FabríCio S. Oliveira

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The agouti is a wildlife rodent that can be used as an alternative source of animal protein and this species has been raised in captivity in Brazil with the aim of providing meat. Thus, the knowledge of their reproductive biology and morphology of the reproductive organs is important. The objective of this study was to describe the morphology of epididymis in the Azara’s agouti, by light microscopy. Samples of epididymis were obtained from five adult Azara’s agouti (Dasyprocta azarae) during castration surgery performed at the Municipal Zoo of Catanduva, Brazil. Fragments of the epididymal regions (initial segment, caput, corpus and cauda) were collected. The biological samples were immediately fixed in paraformaldehyde for 24 hours, followed by histologic procedures comprising embedding in ParaplastTM (Sigma, St. Louis, MO, USA), sections of 5 µm, and staining with HE and Masson’s trichrome. The epididymis was a highly convoluted tubule that links the testis to the vas deferens. The epithelium lining was pseudostratified columnar surrounded by a periductal stroma. The epithelium contains several cell types: principal, basal, apical, clear, and hallo cells. Principal cells were the most abundant cell type. There were observed also migratory cells named halo cells. The caput epididymis was divided into two different regions: initial segment and caput. The initial segment has a very wide lumen, a high epithelium with conspicuous microvilli and the lumen was wide with exfoliated material. The other region of the caput epididymis, showed a lower epithelium when compared with the initial segment, large amounts of spermatozoa in the lumen, and a cytoplasmic vacuolization. This region presented many narrows cells. Many spermatozoa appeared in the lumen of corpus epididymis. The cauda region had a lower epithelium than the other epididymal regions in the agouti. The cauda epithelium presented plicae protruding into the lumen. Large amounts of spermatozoa are also present in the lumen. Small microvilli uniformly arranged so as to form a kind of “brush border” are observed on the apical surface of the cauda epithelium. The pattern of the epithelium lining the duct of the agouti epididymis does not differ greatly from that reported to other mammals, such as domestic and wildlife animals. These findings can cooperate with future investigations especially those related to rational exploration of these animals. All experimental procedures were approved by the institutional ethics committee (CEUA 796/2015). This study was supported by FAPESP (Grants 2015/23822-1).

Keywords: wildlife, testis excurrent ducts, epididymis, morphology

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Authors: Marc Dietrich

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Youth cultures like Punk as much as media relevant to the specific scenes associated with them offer ample opportunity for young people or juvenile adults to construct their personal identities. However, developmental psychology has largely neglected such identity construction processes during the last decades. Such was not always the case: Early developmental psychologists intensely studied youth cultures and their meaningful objects and media in the early 20th century but lost interest when cultural studies and the social sciences occupied the field after World War II. Our project Constructions of Juvenility and Generation(ality), funded by the German Federal Ministry for Education and Research, reintegrates the study of youth cultures and their meaningful objects and media in a developmental psychology perspective. We present an empirical study of the ways in which youth, juvenility, and generation (ality) are constructed and negotiated in underground media like punk fanzines (a portmanteau of fan and magazine), including both semantic and aesthetic aspects of these construction processes within punk culture. The fanzine sample was accessed by the theoretical sampling strategy typical for GTM studies. Acknowledging fanzines as artful self-produced media by scene members for scene members, we conceptualize them as authentic documents of scene norms and values. Drawing on an analysis of both text and (cover) images in Punk fanzines published in Germany (and within a sample dating from 1981 until 2015) using a novel Visual Grounded Theory approach, we found that: a) Juvenility is a highly contested concept in punk culture. Its semantic quality and valuation varies with the perspectives present within the culture (e.g. embryo punks versus older punks); b) Juvenility is constructed as having energy and being socio-critical that does not depend on biological age; c) Juvenility is regarded not an ideal per se in German Punk culture; Punk culture constructs old age in a largely positive way (e.g., as marker of being real and a historical innovator); d) Juvenility is constructed as a habit that should be kept for life as it is constantly adapted to individual biographical trajectories like specific job situations or having a family. Consequently, identity negotiation as documented in the zines attempts to balance subculturally driven perspectives on life and society with the pragmatic requirements of a bourgeois life. The proposed paper will present the main results of this large-scale study of punk fanzines and show how developmental psychology perspectives as represented in the novel methodology applied in it can advance the study of youth cultures.

Keywords: construction of juvenility, developmental psychology, visual GTM, youth culture, fanzines

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Authors: Benoit Atge, Alice Dhersin, Oscar Da Silva Cacao, Beatrice Martinez, Dominique Ducint, Catherine Verdun-Esquer, Isabelle Baldi, Mathieu Molimard, Antoine Villa, Mireille Canal-Raffin

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Introduction: Healthcare workers' exposure to antineoplastic drugs (AD) is a burning issue for occupational medicine practitioners. Biological monitoring of occupational exposure (BMOE) is an essential tool for assessing AD contamination of healthcare workers. In addition to BMOE, surface sampling is a useful tool in order to understand how workers get contaminated, to identify sources of environmental contamination, to verify the effectiveness of surface decontamination way and to ensure monitoring of these surfaces. The objective of this work was to develop a complete tool including a kit for surface sampling and a quantification analytical method for AD traces detection. The development was realized with the three following criteria: the kit capacity to sample in every professional environment (healthcare services, veterinaries, etc.), the detection of very low AD traces with a validated analytical method and the easiness of the sampling kit use regardless of the person in charge of sampling. Material and method: AD mostly used in term of quantity and frequency have been identified by an analysis of the literature and consumptions of different hospitals, veterinary services, and home care settings. The kind of adsorbent device, surface moistening solution and mix of solvents for the extraction of AD from the adsorbent device have been tested for a maximal yield. The AD quantification was achieved by an ultra high-performance liquid chromatography method coupled with tandem mass spectrometry (UHPLC-MS/MS). Results: With their high frequencies of use and their good reflect of the diverse activities through healthcare, 15 AD (cyclophosphamide, ifosfamide, doxorubicin, daunorubicin, epirubicin, 5-FU, dacarbazin, etoposide, pemetrexed, vincristine, cytarabine, methothrexate, paclitaxel, gemcitabine, mitomycin C) were selected. The analytical method was optimized and adapted to obtain high sensitivity with very low limits of quantification (25 to 5000ng/mL), equivalent or lowest that those previously published (for 13/15 AD). The sampling kit is easy to use, provided with a didactic support (online video and protocol paper). It showed its effectiveness without inter-individual variation (n=5/person; n= 5 persons; p=0,85; ANOVA) regardless of the person in charge of sampling. Conclusion: This validated tool (sampling kit + analytical method) is very sensitive, easy to use and very didactic in order to control the chemical risk brought by AD. Moreover, BMOE permits a focal prevention. Used in routine, this tool is available for every intervention of occupational health.

Keywords: surface contamination, sampling kit, analytical method, sensitivity

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Authors: Dalita G. S. M. Cavalcante, Elton A. P. dos Reis, Andressa S. Gomes, Caroline S. Danna, Leandra Ernest Kerche-Silva, Eidi Yoshihara, Aldo E. Job

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In order to minimize environmental impacts, a composite was developed from mixture of leather shavings (LE) with natural rubber (NR), which patent is already deposited. The new material created can be used in applications such as floors e heels for shoes. Besides these applications, the aim is to use this new material for the production of products for the textile industry, such as boots, gloves and bags. But the question arises, as to biocompatibility of this new material. This is justified because the structure of the leather shavings has chrome. The trivalent chromium is usually not toxic, but the hexavalent chromium can be highly toxic and genotoxic for living beings, causing damage to the DNA molecule and contributing to the formation of cancer. Based on this, the objective of this study is evaluate the possible genotoxic effects of the new composite, using as system - test two cell lines (MRC-5 and CHO-K1) by comet assay. For this, the production of the composite was performed in three proportions: for every 100 grams of NR was added 40 (E40), 50 (E50) or 60 (E60) grams of LE. The latex was collected from the rubber tree (Hevea brasiliensis). For vulcanization of the NR, activators and accelerators were used. The two cell lines were exposed to the new composite in its three proportions using elution method, that is, cells exposed to liquid extracts obtained from the composite for 24 hours. For obtaining the liquid extract, each sample of the composite was crushed into pieces and mixed with an extraction solution. The quantification of total chromium and hexavalent chromium in the extracts were performed by Optical Emission Spectrometry by Inductively Coupled Plasma (ICP-OES). The levels of DNA damage in cells exposed to both extracts were monitored by alkaline version of the comet assay. The results of the quantification of metals in ICP-OES indicated the presence of total chromium in different extracts, but were not detected presence of hexavalent chromium in any extract. Through the comet assay were not found DNA damage of the CHO-K1 cells exposed to both extracts. As for MRC-5, was found a significant increase in DNA damage in cells exposed to E50 and E60. Based on the above data, it can be asserted that the extracts obtained from the composite were highly genotoxic for MRC-5 cells. These biological responses do not appear to be related to chromium metal, since there was a predominance of trivalent chromium in the extracts, indicating that during the production process of the new composite, there was no formation of hexavalent chromium. In conclusion it can infer that the leather shavings containing chromium can be reused, thereby reducing the environmental impacts of this waste. Already on the composite indicates to its incorporation in applications that do not aim at direct contact with the human skin, and it is suggested the chain of composite production be studied, in an attempt to make it biocompatible so that it may be safely used by the textile industry.

Keywords: cell line, chrome, genotoxicity, leather, natural rubber

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Authors: Lina Wu

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The excessive discharge of nitrogen in sewage greatly intensifies the eutrophication of water bodies and poses a threat to water quality. Nitrogen pollution control has become a global concern. Currently, the problem of water pollution in China is still not optimistic. As a typical high ammonia nitrogen organic wastewater, landfill leachate is more difficult to treat than domestic sewage because of its complex water quality, high toxicity, and high concentration.Many studies have shown that the autotrophic anammox bacteria in nature can combine nitrous and ammonia nitrogen without carbon source through functional genes to achieve total nitrogen removal, which is very suitable for the removal of nitrogen from leachate. In addition, the process also saves a lot of aeration energy consumption than the traditional nitrogen removal process. Therefore, anammox plays an important role in nitrogen conversion and energy saving. The process composed of short-range nitrification and denitrification coupled an ammo ensures the removal of total nitrogen and improves the removal efficiency, meeting the needs of the society for an ecologically friendly and cost-effective nutrient removal treatment technology. Continuous flow process for treating late leachate [an up-flow anaerobic sludge blanket reactor (UASB), anoxic/oxic (A/O)–anaerobic ammonia oxidation reactor (ANAOR or anammox reactor)] has been developed to achieve autotrophic deep nitrogen removal. In this process, the optimal process parameters such as hydraulic retention time and nitrification flow rate have been obtained, and have been applied to the rapid start-up and stable operation of the process system and high removal efficiency. Besides, finding the characteristics of microbial community during the start-up of anammox process system and analyzing its microbial ecological mechanism provide a basis for the enrichment of anammox microbial community under high environmental stress. One research developed partial nitrification-Anammox (PN/A) using an internal circulation (IC) system and a biological aerated filter (BAF) biofilm reactor (IBBR), where the amount of water treated is closer to that of landfill leachate. However, new high-throughput sequencing technology is still required to be utilized to analyze the changes of microbial diversity of this system, related functional genera and functional genes under optimal conditions, providing theoretical and further practical basis for the engineering application of novel anammox system in biogas slurry treatment and resource utilization.

Keywords: nutrient removal and recovery, leachate, anammox, partial nitrification

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Authors: Carmela Nardelli, Laura Iaffaldano, Valentina Capobianco, Antonietta Tafuto, Maddalena Ferrigno, Angela Capone, Giuseppe Maria Maruotti, Maddalena Raia, Rosa Di Noto, Luigi Del Vecchio, Pasquale Martinelli, Lucio Pastore, Lucia Sacchetti

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Maternal obesity and nutrient excess in utero increase the risk of future metabolic diseases in the adult life. The mechanisms underlying this process are probably based on genetic, epigenetic alterations and changes in foetal nutrient supply. In mammals, the placenta is the main interface between foetus and mother, it regulates intrauterine development, modulates adaptive responses to sub optimal in uterus conditions and it is also an important source of human amniotic mesenchymal stem cells (hA-MSCs). We previously highlighted a specific microRNA (miRNA) profiling in amnion from obese (Ob) pregnant women, here we compared the miRNA expression profile of hA-MSCs isolated from (Ob) and control (Co) women, aimed to search for any alterations in metabolic pathways that could predispose the new-born to the obese phenotype. Methods: We isolated, at delivery, hA-MSCs from amnion of 16 Ob- and 7 Co-women with pre-pregnancy body mass index (mean/SEM) 40.3/1.8 and 22.4/1.0 kg/m2, respectively. hA-MSCs were phenotyped by flow cytometry. Globally, 384 miRNAs were evaluated by the TaqMan Array Human MicroRNA Panel v 1.0 (Applied Biosystems). By the TargetScan program we selected the target genes of the miRNAs differently expressed in Ob- vs Co-hA-MSCs; further, by KEGG database, we selected the statistical significant biological pathways. Results: The immunophenotype characterization confirmed the mesenchymal origin of the isolated hA-MSCs. A large percentage of the tested miRNAs, about 61.4% (232/378), was expressed in hA-MSCs, whereas 38.6% (146/378) was not. Most of the expressed miRNAs (89.2%, 207/232) did not differ between Ob- and Co-hA-MSCs and were not further investigated. Conversely, 4.8% of miRNAs (11/232) was higher and 6.0% (14/232) was lower in Ob- vs Co-hA-MSCs. Interestingly, 7/232 miRNAs were obesity-specific, being expressed only in hA-MSCs isolated from obese women. Bioinformatics showed that these miRNAs significantly regulated (P<0.001) genes belonging to several metabolic pathways, i.e. MAPK signalling, actin cytoskeleton, focal adhesion, axon guidance, insulin signaling, etc. Conclusions: Our preliminary data highlight an altered miRNA profile in Ob- vs Co-hA-MSCs and suggest that an epigenetic miRNA-based mechanism of gene regulation could affect pathways involved in placental growth and function, thereby potentially increasing the newborn’s risk of metabolic diseases in the adult life.

Keywords: hA-MSCs, obesity, miRNA, biosystem

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Authors: Ajuna B. Henry

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In recent decades, climate change has demanded biological pesticides; more Bt strains are being discovered worldwide, some containing novel insecticidal genes while others have been modified through molecular approaches for increased yield, toxicity, and wider host target. In this study, B. thuringiensis strain AH-2 (Bt-2) was isolated from the soil and tested for insecticidal activity against Aphis gossypii (Hemiptera: Aphididae) and Lepidoptera insect pests: fall webworm (Hyphantria cunea) and diamondback moth (Plutella xylostella). A commercial strain B. thuringiensis subsp. kurstaki (Btk), and a chemical pesticide, imidacloprid (for Hemiptera) and chlorantraniliprole (for Lepidoptera), were used as positive control and the same media (without bacterial inoculum) as a negative control. For aphidicidal activity, Bt-2 caused a mortality rate of 70.2%, 78.1% or 88.4% in third instar nymphs of A. gossypii (3N) at 10%, 25% or 50% culture concentrations, respectively. Moreover, Bt-2 was effectively produced in cost-effective (PB) supplemented with either glucose (PBG) or sucrose (PBS) and maintained high aphicidal efficacy with 3N mortality rates of 85.9%, 82.9% or 82.2% in TSB, PBG or PBS media, respectively at 50% culture concentration. Bt-2 also suppressed adult fecundity by 98.3% compared to only 65.8% suppression by Btk at similar concentrations but was slightly lower than chemical treatment, which caused 100% suppression. Partial purification of 60 – 80% (NH4)2SO4 fraction of Bt-2 aphicidal proteins purified on anion exchange (DEAE-FF) column revealed a 105 kDa aphicidal protein with LC50 = 55.0 ng/µℓ. For Lepidoptera pests, chemical pesticide, Bt-2, and Btk cultures, mortality of 86.7%, 60%, and 60% in 3rd instar larvae of P. xylostella, and 96.7%, 80.0%, and 93.3% in 6th instar larvae of H. cunea, after 72h of exposure. When the entomopathogenic strains were cultured in a cost-effective PBG or PBS, the insecticidal activity in all strains was not significantly different compared to the use of a commercial medium (TSB). Bt-2 caused a mortality rate of 60.0%, 63.3%, and 50.0% against P. xylostella larvae and 76.7%, 83.3%, and 73.3% against H. cunea when grown in TSB, PBG, and PBS media, respectively. Bt-2 (grown in cost-effective PBG medium) caused a dose-dependent toxicity of 26.7%, 40.0%, and 63.3% against P. xylostella and 46.7%, 53.3%, and 76.7% against H. cunea at 10%, 25% and 50% culture concentration, respectively. The partially purified Bt-2 insecticidal proteins fractions F1, F2, F3, and F4 (extracted at different ratios of organic solvent) caused low toxicity (50.0%, 40.0%, 36.7%, and 30.0%) against P. xylostella and relatively high toxicity (56.7%, 76.7%, 66.7%, and 63.3%) against H. cunea at 100 µg/g of artificial diets. SDS-PAGE analysis revealed that a128kDa protein is associated with toxicity of Bt-2. Our result demonstrates a medium and strong larvicidal activity of Bt-2 against P. xylostella and H. cunea, respectively. Moreover, Bt-2 could be potentially produced using a cost-effective PBG medium which makes it an effective alternative biocontrol strategy to reduce chemical pesticide application.

Keywords: biocontrol, insect pests, larvae/nymph mortality, cost-effective media, aphis gossypii, plutella xylostella, hyphantria cunea, bacillus thuringiensi

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Authors: Belcher Fulele, W. A. A. Ddamba

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Study of solvent systems contributes to the understanding of intermolecular interactions that occur in binary mixtures. These interactions involves among others strong dipole-dipole interactions and weak van de Waals interactions which are of significant application in pharmaceuticals, solvent extractions, design of reactors and solvent handling and storage processes. Binary mixtures of solvents can thus be used as a model to interpret thermodynamic behavior that occur in a real solution mixture. Densities of pure DFM, n-alkanes (n-pentane, n-hexane, n-heptane and n-octane) and amides (N-methylformamide, N-ethylformamide, N,N-dimethylformamide and N,N-dimethylacetamide) as well as their [DFM + ((C5-C8) n-alkane or amide)] binary mixtures over the entire composition range, have been reported at temperature 293.15, 298.15 and 303.15 K and atmospheric pressure. These data has been used to derive the thermodynamic properties: the excess molar volume of solution, apparent molar volumes, excess partial molar volumes, limiting excess partial molar volumes, limiting partial molar volumes of each component of a binary mixture. The results are discussed in terms of possible intermolecular interactions and structural effects that occur in the binary mixtures. The variation of excess molar volume with DFM composition for the [DFM + (C5-C7) n-alkane] binary mixture exhibit a sigmoidal behavior while for the [DFM + n-octane] binary system, positive deviation of excess molar volume function was observed over the entire composition range. For each of the [DFM + (C5-C8) n-alkane] binary mixture, the excess molar volume exhibited a fall with increase in temperature. The excess molar volume for each of [DFM + (NMF or NEF or DMF or DMA)] binary system was negative over the entire DFM composition at each of the three temperatures investigated. The negative deviations in excess molar volume values follow the order: DMA > DMF > NEF > NMF. Increase in temperature has a greater effect on component self-association than it has on complex formation between molecules of components in [DFM + (NMF or NEF or DMF or DMA)] binary mixture which shifts complex formation equilibrium towards complex to give a drop in excess molar volume with increase in temperature. The Prigogine-Flory-Patterson model has been applied at 298.15 K and reveals that the free volume is the most important contributing term to the excess experimental molar volume data for [DFM + (n-pentane or n-octane)] binary system. For [DFM + (NMF or DMF or DMA)] binary mixture, the interactional term and characteristic pressure term contributions are the most important contributing terms in describing the sign of experimental excess molar volume. The mixture systems contributed to the understanding of interactions of polar solvents with proteins (amides) with non-polar solvents (alkanes) in biological systems.

Keywords: alkanes, amides, excess thermodynamic parameters, Prigogine-Flory-Patterson model

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Authors: Fanny Desbiolles, Laure Malleret, Isabelle Laffont-Schwob, Christophe Tiliacos, Anne Piram, Mohamed Sarakha, Pascal Wong-Wah-Chung

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Pharmaceuticals in the environment have become a very hot topic in the recent years. This interest is related to the large amounts dispensed and to their release in urine or faeces from treated patients, resulting in their ubiquitous presence in water resources and wastewater treatment plants (WWTP) effluents. Thereby, many studies focused on the prediction of pharmaceuticals’ behaviour, to assess their fate and impacts in the environment. Carbamazepine is a widely consumed psychotropic pharmaceutical, thus being one of the most commonly detected drugs in the environment. This organic pollutant was proved to be persistent, especially with respect to its non-biodegradability, rendering it recalcitrant to usual biological treatment processes. Consequently, carbamazepine is very little removed in WWTP with a maximum abatement rate of 5 % and is then often released in natural surface waters. To better assess the environmental fate of carbamazepine in aqueous media, its photochemical transformation was undertaken in four natural waters (two French rivers, the Berre salt lagoon, Mediterranean Sea water) representative of coastal and inland water types. Kinetic experiments were performed in the presence of light using simulated solar irradiation (Xe lamp 300W). Formation of short-lifetime species was highlighted using chemical trap and laser flash photolysis (nanosecond). Identification of transformation by-products was assessed by LC-QToF-MS analyses. Carbamazepine degradation was observed after a four-day exposure and an abatement of 20% maximum was measured yielding to the formation of many by-products. Moreover, the formation of hydroxyl radicals (•OH) was evidenced in waters using terephthalic acid as a probe, considering the photochemical instability of its specific hydroxylated derivative. Correlations were implemented using carbamazepine degradation rate, estimated hydroxyl radical formation and chemical contents of waters. In addition, laser flash photolysis studies confirmed •OH formation and allowed to evidence other reactive species, such as chloride (Cl2•-)/bromine (Br2•-) and carbonate (CO3•-) radicals in natural waters. Radicals mainly originate from dissolved phase and their occurrence and abundance depend on the type of water. Rate constants between reactive species and carbamazepine were determined by laser flash photolysis and competitive reactions experiments. Moreover, LC-QToF-MS analyses of by-products help us to propose mechanistic pathways. The results will bring insights to the fate of carbamazepine in various water types and could help to evaluate more precisely potential ecotoxicological effects.

Keywords: carbamazepine, kinetic and mechanistic approaches, natural waters, photodegradation

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Authors: Tomas Makaras, Gintaras Svecevičius

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Landfill waste is a common problem as it has an economic and environmental impact even if it is closed. Landfill waste contains a high density of various persistent compounds such as heavy metals, organic and inorganic materials. As persistent compounds are slowly-degradable or even non-degradable in the environment, they often produce sublethal or even lethal effects on aquatic organisms. The aims of the present study were to estimate sublethal effects of the Kairiai landfill (WGS: 55°55‘46.74“, 23°23‘28.4“) leachate on the locomotor activity of rainbow trout Oncorhynchus mykiss juveniles using the original system package developed in our laboratory for automated monitoring, recording and analysis of aquatic organisms’ activity, and to determine patterns of fish behavioral response to sublethal effects of leachate. Four different concentrations of leachate were chosen: 0.125; 0.25; 0.5 and 1.0 mL/L (0.0025; 0.005; 0.01 and 0.002 as part of 96-hour LC50, respectively). Locomotor activity was measured after 5, 10 and 30 minutes of exposure during 1-minute test-periods of each fish (7 fish per treatment). The threshold-effect-concentration amounted to 0.18 mL/L (0.0036 parts of 96-hour LC50). This concentration was found to be even 2.8-fold lower than the concentration generally assumed to be “safe” for fish. At higher concentrations, the landfill leachate solution elicited behavioral response of test fish to sublethal levels of pollutants. The ability of the rainbow trout to detect and avoid contaminants occurred after 5 minutes of exposure. The intensity of locomotor activity reached a peak within 10 minutes, evidently decreasing after 30 minutes. This could be explained by the physiological and biochemical adaptation of fish to altered environmental conditions. It has been established that the locomotor activity of juvenile trout depends on leachate concentration and exposure duration. Modeling of these parameters showed that the activity of juveniles increased at higher leachate concentrations, but slightly decreased with the increasing exposure duration. Experiment results confirm that the behavior of rainbow trout juveniles is a sensitive and rapid biomarker that can be used in combination with the system for fish behavior monitoring, registration and analysis to determine sublethal concentrations of pollutants in ambient water. Further research should be focused on software improvement aimed to include more parameters of aquatic organisms’ behavior and to investigate the most rapid and appropriate behavioral responses in different species. In practice, this study could be the basis for the development and creation of biological early-warning systems (BEWS).

Keywords: fish behavior biomarker, landfill leachate, locomotor activity, rainbow trout juveniles, sublethal effects

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Authors: Agne Jucyte Cicine, Vytautas Abromaitis, Zita Rasuole Gasiunaite, I. Vybernaite-Lubiene, D. Overlinge, K. Vilke

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Pharmaceutical pollution in aquatic environments has become a growing concern. Various active pharmaceutical ingredient (API) residues, hormones, antibiotics, or/and psychiatric drugs, have already been discovered in different environmental compartments. Due to existing ineffective wastewater treatment technologies to remove APIs, an underestimated amount can enter the ecosystem by discharged treated wastewater. Especially, psychiatric compounds, such as carbamazepine (CBZ) and venlafaxine (VNX), persist in effluent even post-treatment. Therefore, these pharmaceuticals usually exceed safe environmental levels and pose risks to the aquatic environment, particularly to sensitive ecosystems such as the Baltic Sea. CBZ, known for its chemical stability and long biodegradation time, accumulates in the environment, threatening aquatic life and human health through the food chain. As the use of medication rises, there is an urgent need for advanced wastewater treatment to reduce pharmaceutical contamination and meet future regulatory requirements. In this study, we tested advanced oxidation technology using ozone to remove two commonly used psychiatric drugs (carbamazepine and venlafaxine) from biologically treated wastewater effluent. Additionally, general water quality parameters (suspended matter (SPM), dissolved organic carbon (DOC), chemical oxygen demand (COD), and bacterial presence were analyzed. Three wastewater treatment plants (WWTPs) representing different anthropogenic pressures were selected: 1) resort, 2) resort and residential, and 3) residential, industrial, and resort. Wastewater samples for the experiment were collected during the summer season after mechanical and biological treatment and ozonated for 5, 10, and 15 minutes. The initial dissolved ozone concentration of 7,3±0,7 mg/L was held constant during all the experiments. Pharmaceutical levels in this study exceeded the predicted no-effect concentration (PNEC) of 500 and 90 ng L⁻¹ for CBZ and VNX, respectively, in all WWTPs, except CBZ in WWTP 1. Initial CBZ contamination was found to be lower in WWTP 1 (427.4 ng L⁻¹), compared with WWTP 2 (1266.5 ng L⁻¹) and 3 (119.2 ng L⁻¹). VNX followed a similar trend with concentrations of 341.2 ng L⁻¹, 361.4 ng L⁻¹, and 390.0 ng L⁻¹, respectively, for WWTPs 1, 2, and 3. It was determined that CBZ was not detected in the effluent after 5 minutes of ozonation in any of the WWTPs. Contrarily, VNX was still detected after 5, 10, and 15 minutes of treatment with ozone, however, under the limits of quantification (LOD) (<5ng L⁻¹). Additionally, general pollution of SPM, DOC, COD, and bacterial contamination was reduced notably after 5 minutes of treatment with ozone, while no bacterial growth was obtained. Although initial pharmaceutical levels exceeded PNECs, indicating ongoing environmental risks, ozonation demonstrated high efficiency in reducing pharmaceutical and general contamination in wastewater with different pollution matrices.

Keywords: Baltic Sea, ozonation, pharmaceuticals, wastewater treatment plants

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Authors: Izabela Poprawa, Alina Chachulska-Zymelka, Lukasz Chajec, Grazyna Wilczek, Piotr Wilczek, Sebastian Student, Magdalena Rost-Roszkowska

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Heavy metals polluting the environment, especially soil, have a harmful effect on organisms, because they can damage the organ structure, disturb their function and cause developmental disorders. They can affect not only the somatic tissues but also the germinal tissues. In the natural environment, plants and animals are exposed to short- and long-term exposure to these stressors, which have a major influence on the functioning of these organisms. Numerous animals have been treated as the bioindicators of the environment. Therefore, studies on any alterations caused by, e.g., heavy metals are in the center of interests of not only environmental but also medical and biological science. Myriapods are invertebrates which are bioindicators of the environment. One of the species which lives in the upper layers of soil, particularly under stones and rocks is Lithobius forficatus (Chilopoda), commonly known as the brown centipede or stone centipede. It is a European species of the family Lithobiidae. This centipede living in the soil is exposed to, e.g., heavy metals such as cadmium, lead, arsenic. The main goal of our project was to analyze the impact of long-term exposure to cadmium on the structure of ovary with the emphasis on the course of oogenesis. As the material for analysis of cadmium exposure to ovaries, we chose the centipede species, L. forficatus. Animals were divided into two experimental groups: C – the control group, the animals cultured in laboratory conditions in a horticultural soil; Cd2 – the animals cultured in a horticultural soil supplemented with 80 mg/kg (dry weight) of CdCl2 for 45 days – long-term exposure. Animals were fed with Acheta and Chironomus larvae maintained in tap water. The analyzes were carried out using transmission electron microscopy (TEM), flow cytometry and laser scanning (confocal) microscopy. Here we present the results of long-term exposure to cadmium concentration in soil on the organ responsible for female germ cell formation. Analysis with the use of the transmission electron microscope showed changes in the ultrastructure of both somatic and germ cells in the ovary. Moreover, quantitative analysis revealed the decrease in the percentage of cells viability, the increase in the percentage of cells with depolarized mitochondria and increasing the number of early apoptotic cells. All these changes were statistically significant compared to the control. Additionally, an increase in the ADP/ATP index was recorded. However, changes were not statistically significant to the control. Acknowledgment: The study has been financed by the National Science Centre, Poland, grant no 2017/25/B/NZ4/00420.

Keywords: cadmium, centipede, ovary, ultrastructure

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Authors: Vanessa Bennemann, Gabriela Laste, Márcia Inês Goettert

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The gestational period is a phase in which the pregnant woman undergoes constant physiological and hormonal changes, which are part of the woman’s biological cycle, the development of the fetus, childbirth, and lactation. These are factors of response to the immunological adaptation of the human reproductive process that is directly related to the pregnancy’s well-being and development. Although most pregnancies occur without complications, about 15% of pregnant women will develop potentially fatal complications, implying maternal and fetal risk. Therefore, requiring specialized care for high-risk pregnant women (HRPW) with obstetric interventions for the survival of the mother and/or fetus. Among the risk factors that characterize HRPW are the women's age, gestational diabetes mellitus (GDM), autoimmune diseases, infectious diseases such as syphilis and HIV, hypertension (SAH), preeclampsia, eclampsia, HELLP syndrome, uterine contraction abnormalities, and premature placental detachment (PPD), thyroid disorders, among others. Thus, pregnancy has an impact on the thyroid gland causing changes in the functioning of the mother's thyroid gland, altering the thyroid hormone (TH) profiles and production as pregnancy progresses. Considering, throughout the gestational period, the interpretation of the results of the tests to evaluate the thyroid functioning depends on the stage in which the pregnancy is. Thyroid disorders are directly related to adverse obstetric outcomes and in child development. Therefore, the adequate release of TH is important for a pregnancy without complications and optimal fetal growth and development. Objective: Investigate the physiological effects caused by thyroid disorders in the gestational period. Methods: A search for articles indexed in PubMed, Scielo, and MDPI databases, was performed using the term “AND”, with the descriptors: Pregnancy, Thyroid. With several combinations that included: Melatonin, Thyroidopathy, Inflammatory processes, Cytokines, Anti-inflammatory, Antioxidant, High-risk pregnancy. Subsequently, the screening was performed through the analysis of titles and/or abstracts. The criteria were: including clinical studies in general, randomized or not, in the period of 10 years prior to the research, in the English literature; excluded: experimental studies, case reports, research in the development phase. Results: In the preliminary results, a total of studies (n=183) were found, (n=57) excluded, such as studies of cancer, diabetes, obesity, and skin diseases. Conclusion: To date, it has been identified that thyroid diseases can impair the fetus’s brain development. Further research is suggested on this matter to identify new substances that may have a potential therapeutic effect to aid the gestational period with thyroid diseases.

Keywords: pregnancy, thyroid, melatonin, high-risk pregnancy

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Authors: Muhammad Fiaz Qamar, Uzma Mehreen, Muhammad Arfan Zaman, Kazim Ali

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Ovine Theileriosis is a world-wide concern, especially in tropical and subtropical areas, due to having tick abundance that has received less awareness in different developed and developing areas due to less worth of sheep, low to the middle level of infection in different small ruminants herd. Across Asia, the prevalence reports have been conducted to provide equivalent calculation of flock and animal level prevalence of Theileriosisin animals. It is a challenge for veterinarians to timely diagnosis & control of Theileriosis and famers because of the nature of the organism and inadequacy of restricted plans to control. All most work is based upon the development of such a technique which should be farmer-friendly, less expensive, and easy to perform into the field. By the timely diagnosis of this disease will decrease the irrational use of the drugs, and other plan was to determine the prevalence of Theileriosis in District Jhang by using the conventional method, PCR and qPCR, and LAMP. We quantify the molecular epidemiology of T.lestoquardiin sheep from Jhang districts, Punjab, Pakistan. In this study, we concluded that the overall prevalence of Theileriosis was (32/350*100= 9.1%) in sheep by using Giemsa staining technique, whereas (48/350*100= 13%) is observed by using PCR technique (56/350*100=16%) in qPCR and the LAMP technique have shown up to this much prevalence percentage (60/350*100= 17.1%). The specificity and sensitivity also calculated in comparison with the PCR and LAMP technique. Means more positive results have been shown when the diagnosis has been done with the help of LAMP. And there is little bit of difference between the positive results of PCR and qPCR, and the least positive animals was by using Giemsa staining technique/conventional method. If we talk about the specificity and sensitivity of the LAMP as compared to PCR, The cross tabulation shows that the results of sensitivity of LAMP counted was 94.4%, and specificity of LAMP counted was 78%. Advances in scientific field must be upon reality based ideas which can lessen the gaps and hurdles in the way of scientific research; the lamp is one of such techniques which have done wonders in adding value and helping human at large. It is such a great biological diagnostic tools and has helped a lot in the proper diagnosis and treatment of certain diseases. Other methods for diagnosis, such as culture techniques and serological techniques, have exposed humans with great danger. However, with the help of molecular diagnostic technique like LAMP, exposure to such pathogens is being avoided in the current era Most prompt and tentative diagnosis can be made using LAMP. Other techniques like PCR has many disadvantages when compared to LAMP as PCR is a relatively expensive, time consuming, and very complicated procedure while LAMP is relatively cheap, easy to perform, less time consuming, and more accurate. LAMP technique has removed hurdles in the way of scientific research and molecular diagnostics, making it approachable to poor and developing countries.

Keywords: distribution, thelaria, LAMP, primer sequences, PCR

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Authors: Ming-Yeh Lu, Shiao-Shing Chen, Saikat Sinha Ray, Hung-Te Hsu

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Recently, anaerobic membrane bioreactors (AnMBRs) has been widely utilized, which combines anaerobic biological treatment process and membrane filtration, that can be present an attractive option for wastewater treatment and water reuse. Conventional AnMBR is having several advantages, such as improving effluent quality, compact space usage, lower sludge yield, without aeration and production of energy. However, the removal of nitrogen and phosphorus in the AnMBR permeate was negligible which become the biggest disadvantage. In recent years, forward osmosis (FO) is an emerging technology that utilizes osmotic pressure as driving force to extract clean water without additional external pressure. The pore size of FO membrane is kindly mentioned the pore size, so nitrogen or phosphorus could effectively improve removal of nitrogen or phosphorus. Anaerobic bioreactor with FO membrane (AnOMBR) can retain the concentrate organic matters and nutrients. However, phosphorus is a non-renewable resource. Due to the high rejection property of FO membrane, the high amount of phosphorus could be recovered from the combination of AnMBR and FO. In this study, development of novel submerged anaerobic osmotic membrane bioreactor integrated with periodic microfiltration (MF) extraction for simultaneous phosphorus and clean water recovery from wastewater was evaluated. A laboratory-scale AnOMBR utilizes cellulose triacetate (CTA) membranes with effective membrane area of 130 cm² was fully submerged into a 5.5 L bioreactor at 30-35℃. Active layer-facing feed stream orientation was utilized, for minimizing fouling and scaling. Additionally, a peristaltic pump was used to circulate draw solution (DS) at a cross flow velocity of 0.7 cm/s. Magnesium sulphate (MgSO₄) solution was used as DS. Microfiltration membrane periodically extracted about 1 L solution when the TDS reaches to 5 g/L to recover phosphorus and simultaneous control the salt accumulation in the bioreactor. During experiment progressed, the average water flux was achieved around 1.6 LMH. The AnOMBR process show greater than 95% removal of soluble chemical oxygen demand (sCOD), nearly 100% of total phosphorous whereas only partial removal of ammonia, and finally average methane production of 0.22 L/g sCOD was obtained. Therefore, AnOMBR system periodically utilizes MF membrane extracted for phosphorus recovery with simultaneous pH adjustment. The overall performance demonstrates that a novel submerged AnOMBR system is having potential for simultaneous wastewater treatment and resource recovery from wastewater, and hence, the new concept of this system can be used to replace for conventional AnMBR in the future.

Keywords: anaerobic treatment, forward osmosis, phosphorus recovery, membrane bioreactor

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Authors: I. Nava-Arenas, N. Ruiz-Ordaz, C. J. Galindez-Mayer, M. L. Luna-Guido, S. L. Ruiz-López, A. Cabrera-Orozco, D. Nava-Arenas

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Among the most important herbicides, the organochlorine compounds are of considerable interest due to their recalcitrance to the chemical, biological, and photolytic degradation, their persistence in the environment, their mobility, and their bioacummulation. The most widely used herbicides in North America are primarily 2,4-dichlorophenoxyacetic acid (2,4-D), the triazines (atrazine and simazine), and to a lesser extent diuron. The contamination of soils and water bodies frequently occurs by mixtures of these xenobiotics. For this reason, in this work, the operational stability to changes in the composition of the medium supplied to an aerobic biofilm reactor was studied. The reactor was packed with fragments of volcanic rock that retained a complex microbial film, able to degrade a mixture of organochlorine herbicides atrazine, simazine, diuron and 2,4-D, and whose members have microbial genes encoding the main catabolic enzymes atzABCD, tfdACD and puhB. To acclimate the attached microbial community, the biofilm reactor was fed continuously with a mineral minimal medium containing the herbicides (in mg•L-1): diuron, 20.4; atrazine, 14.2, simazine, 11.4, and 2,4-D, 59.7, as carbon and nitrogen sources. Throughout the bioprocess, removal efficiencies of 92-100% for herbicides, 78-90% for COD, 92-96% for TOC and 61-83% for dehalogenation were reached. In the microbial community, the genes encoding catabolic enzymes of different herbicides tfdACD, puhB and, occasionally, the genes atzA and atzC were detected. After the acclimatization, the triazine herbicides were eliminated from the mixture formulation. Volumetric loading rates of the mixture 2,4-D and diuron were continuously supplied to the reactor (1.9-21.5 mg herbicides •L-1 •h-1). Along the bioprocess, the removal efficiencies obtained were 86-100% for the mixture of herbicides, 63-94% for for COD, 90-100% for COT, and dehalogenation values of 63-100%. It was also observed that the genes encoding the enzymes in the catabolism of both herbicides, tfdACD and puhB, were consistently detected; and, occasionally, the atzA and atzC. Subsequently, the triazine herbicide atrazine and simazine were restored to the medium supply. Different volumetric charges of this mixture were continuously fed to the reactor (2.9 to 12.6 mg herbicides •L-1 •h-1). During this new treatment process, removal efficiencies of 65-95% for the mixture of herbicides, 63-92% for COD, 66-89% for TOC and 73-94% of dehalogenation were observed. In this last case, the genes tfdACD, puhB and atzABC encoding for the enzymes involved in the catabolism of the distinct herbicides were consistently detected. The atzD gene, encoding the cyanuric hydrolase enzyme, could not be detected, though it was determined that there was partial degradation of cyanuric acid. In general, the community in the biofilm reactor showed some catabolic stability, adapting to changes in loading rates and composition of the mixture of herbicides, and preserving their ability to degrade the four herbicides tested; although, there was a significant delay in the response time to recover to degradation of the herbicides.

Keywords: biodegradation, biofilm reactor, microbial community, organochlorine herbicides

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Authors: D. Scarpanti, M. Brondino, M. Pasini

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Background: the present study explores the role of light and restorativeness on work. According with the Attention Restoration Theory (ART) and a Model of Work Environment, the main idea is that some features of environment, i.e., lighting, influences the direct attention, and so, the performance. Restorativeness refers to the presence/absence level of all the characteristics of physical environment that help to regenerate direct attention. Specifically, lighting can affect level of fascination and attention in one hand; and in other hand promotes several biological functions via pineal gland. Different reviews on this topic show controversial results. In order to bring light on this topic, the hypotheses of this study are that lighting can affect the construct of restorativeness and, in the second time, the restorativeness can affect the performance. Method: the participants are 30 workers of a mechatronic company in the North Italy. Every subject answered to a questionnaire valuing their subjective perceptions of environment in a different way: some objective features of environment, like lighting, temperature and air quality; some subjective perceptions of this environment; finally, the participants answered about their perceived performance. The main attention is on the features of light and his components: visual comfort, general preferences and pleasantness; and the dimensions of the construct of restorativeness; fascination, coherence and being away. The construct of performance per se is conceptualized in three level: individual, team membership and organizational membership; and in three different components: proficiency, adaptability, and proactivity, for a total of 9 subcomponents. Findings: path analysis showed that some characteristics of lighting respectively affected the dimension of fascination; and, as expected, the dimension of fascination affected work performance. Conclusions: The present study is a first pilot step of a wide research. These first results can be summarized with the statement that lighting and restorativeness contribute to explain work performance variability: in details perceptions of visual comfort, satisfaction and pleasantness, and fascination respectively. Results related to fascination are particularly interesting because fascination is conceptualized as the opposite of the construct of direct attention. The main idea is, in order to regenerate attentional capacity, it’s necessary to provide a lacking of attention (fascination). The sample size did not permit to test simultaneously the role of the perceived characteristics of light to see how they differently contribute to predict fascination of the work environment. However, the results highlighted the important role that light could have in predicting restorativeness dimensions and probably with a larger sample we could find larger effects also on work performance. Furthermore, longitudinal data will contribute to better analyze the causal model along time. Applicative implications: the present pilot study highlights the relevant role of lighting and perceived restorativeness in the work environment and the importance to focus attention on light features and the restorative characteristics in the design of work environments.

Keywords: lighting, performance, restorativeness, workplace

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Authors: Merav Meiron-Goren, Orna Braun-Lewensohn, Tal Litvak-Hirsh

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In recent years, there has been an increasing tendency towards separation and divorce in relationships. Nevertheless, many couples in a first marriage do not anticipate this as a probable possibility and do not make any preparation for it. Same sex couples establishing a family encounter a much more complicated situation than do heterosexual couples. Although there is a trend towards legal recognition of same sex marriage, many countries, including Israel, do not recognize it. The absence of legal recognition or the existence of partial recognition creates complexity for these couples. They have to fight for their right to establish a family, like the recognition of the biological child of a woman, as a child of her woman spouse too, or the option of surrogacy for a male couple who want children, and more. The lack of legal recognition is burden on the lives of these couples. In the absence of clear norms regarding the conduct of the family unit, the couples must define for themselves the family structure, and deal with everyday dilemmas that lack institutional solutions. This may increase the friction between the two couple members, and it is one of the factors that make it difficult for them to maintain the relationship. This complexity exists, perhaps even more so, in separation. The end of relationship is often accompanied by a deep crisis, causing pain and stress. In most cases, there are also other conflicts that must be settled. These are more complicated when rights are in doubt or do not exist at all. Complex issues for separating same sex couples may include matters of property, recognition of parenthood, and care and support for the children. The significance of the study is based on the fact that same sex relationships are becoming more and more widespread, and are an integral part of the society. Even so, there is still an absence of research focusing on such relationships and their ending. The objective of the study is to research the perceptions of same sex couples regarding the possibility of separation, preparing for it, conflict management and resolving disputes through the separation process. It is also important to understand the point of view of couples that have gone through separation, how they coped with the emotional and practical difficulties involved in the separation process. The doctoral research will use a qualitative research method in a phenomenological approach, based on semi-structured in-depth interviews. The interviewees will be divided into three groups- at the beginning of a relationship, during the separation crisis and after separation, with a time perspective, with about 10 couples from each group. The main theoretical model serving as the basis of the study will be the Lazarus and Folkman theory of coping with stress. This model deals with the coping process, including cognitive appraisal of an experience as stressful, appraisal of the coping resources, and using strategies of coping. The strategies are divided into two main groups, emotion-focused forms of coping and problem-focused forms of coping.

Keywords: conflict management, coping, legal recognition, same-sex relationship, separation

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