Light Period Research Articles

Energy Storage System Owner as a New Player in an Electricity Structure

Daily load demand causes different marginal cost at each hour due to different operation generation units. During peak load period, expensive generation units are determined to be turned on to provide sufficient electricity supply and sufficient spinning reserve. During light load period, generation units could not be unloaded due to their minimum up/down time, startup time and startup cost, causing uneconomic operation of the generating capacities. This excess capacity during light load period can be stored in the energy storage system (ESS) and the power can be released to supply the peak load demand hours to avoid turning on the next expensive unit which resulted in higher marginal cost. Performing as virtual power plant (VPP), ESS owners can seek for market opportunities to enter electric supply industry. This paper aims for proposing an optimal operation of VPP with charging/discharging ESS plan when marginal cost identifies real-time pricing (RTP) at each hour to be used as buying/selling price to VPP. Under strategically charging/discharging scheme, both parties, i.e., utility, and VPP, can achieve benefits in terms of better economic operation, lower system generation cost, increase operational income, while environmental impact is considered. The proposed method is tested on a ten-unit system under a centralized power market structure. Numerical results show that appropriate charging/discharging strategy could provide lower total production cost and offer opportunities for ESS owners as VPPs to obtain arbitrage marginal cost.

Ecological benefits of artificial light at night (ALAN): Accelerating the development and metamorphosis of marine shellfish larvae

Urbanization has led to increasing use of artificial light at night (ALAN), which has rapidly become an important source of pollution in many cities. To identify the ALAN effects on the embryonic development of the Pacific abalone Haliotis discus hannai, we first exposed larvae to natural light with a light period of 12 L:12D (control, Group CTR). We then exposed larvae to three different light regimes. Larvae in Group NL were exposed to full spectrum artificial light from 18:00 to 00:00 to simulate the lighting condition at night, whereas Groups BL and YL were illuminated at the same time interval with 450 nm of short-wavelength blue light and 560 nm of long-wavelength orange light, respectively, to simulate billboard lighting at night. There were significantly higher hatching success and metamorphosis rates of larvae in Group BL than in Group YL or CTR (P < 0.05). The larvae in Group YL had the highest abnormality rate and took the longest time to complete metamorphosis. Transcriptomic studies revealed significantly higher expression levels of genes related to RNA transport, DNA replication, and protein processing in endoplasmic reticulum pathways in Group BL compared to the other groups. In the metabolomic analysis, we identified prostaglandin B1, tyramine, d-fructose 6-phosphate, L-adrenaline, leukotriene C4, and arachidonic acid as differential metabolic markers, as they play a vital part in helping larvae adapt to different ALAN conditions. Multi-omics correlation analysis of pairwise comparisons between all of the groups suggested that the biosynthesis of unsaturated fatty acids (FAs) and arachidonic acid metabolism pathways were significantly enriched (P < 0.05). Further quantitative analysis of the fatty acid (FA) contents revealed that 42 out of 50 FAs were down-regulated in Group BL and up-regulated in Group YL, which suggested that the synthesis, catabolism, and metabolism of FAs are crucial for the larval response to different spectral components of ALAN. For the first time, we report positive rather than negative effects of artificial blue light at night on the embryonic development of a benthic marine species. These results are significant for unbiased and full-scale assessment of the ecological effects of ALAN and for understanding the structural stability of the marine benthic community.

Behavioral changes of food allergic model mice during light and dark period

Although an animal model of food allergy has been used to investigate its progression mechanism, most researcher could not assess its symptoms for long especially under dark environment. We assessed the behavioral changes of food allergic mice using an image analysis system to track a mouse under both light and dark environments. Mice were sensitized with intraperitoneal ovalbumin (OVA) injections and challenged ten times with oral OVA administration. The OVA challenges induced weight loss and diarrhea. We assessed their behavior and found that the OVA challenges decreased their total moving distance during the dark period. We also revealed that the OVA challenges increased the inactive time of mice during the dark period. Interestingly, these changes were not observed or very small during the light period. We next assessed the location of mice in the home-cage and found that the OVA challenges increased the time when mice stayed at corners and decreased the time at the center during the dark period. These observations suggest mental abnormality of mice. Indeed, the OVA challenges increased the immobility time of mice in the tail suspension test. Thus, food allergic mice exhibited reduced activity and might exhibit psychological symptoms during dark period.

First report of Neopestalotiopsis clavispora causing cashew leaf blight disease in India.

Cashew (Anacardium occidentale) is an important commercial crop and highly prone to many biotic and abiotic stress. During March 2021, severe leaf blight symptoms were observed in Priyanka variety with 25-30% incidence grown under greenhouse nursery at ICAR-Directorate of Cashew Research (ICAR-DCR), Puttur (12º74'08.92"N; 75º22'97.22"E), Karnataka. Initial symptoms include small, irregular necrotic spots and later, the spots enlarged and covered major portion of the leaf lamina. In severe infection, leaves exhibited coalescing of spots leading to blight appearance. The infected leaves were randomly collected (n=5) and surface sterilized with 1% sodium hypochlorite for 1 min followed by three washes in sterile distilled water (SDW). Samples were plated on PDA plates amended with Rifampicin (40 mg/L) and kept for incubation at 25±2 oC for 5 days (12/12 h dark light period). A white-greyish, aerial, cottony mycelium on upper side with light yellow colour on the reverse side was consistently isolated. The black viscous acervuli were observed after 10-12 days of incubation. The conidia were fusiform, five-celled, versicoloured with three olivaceous brown median cells, two terminal hyaline cells, measured 23.3±2.12 - 28.33±2.7 x 3.6±0.8 - 4.28±0.78 µm (n=30). The apical cells had two to three flexuous, unbranched appendages, and basal appendage was solitary, tubular and unbranched. Morphological and cultural characteristics confirmed the pathogen as Neopestalotiopsis sp. (Maharachchikumbura et al. 2012). Further, two representative isolates (CLB_SCN1 & CLB_SCN2) were subjected for molecular characterization selected for molecular identification based on ITS-rDNA, tef-1α and tub2 gene sequences and phylogenetic analysis. Genomic DNA was isolated from 15 days old cultures and internal transcribed spacer (ITS) of ribosomal DNA (rDNA) (White et al. 1990), translation elongation factor 1α (tef-1α) gene (O'Donnell et al. 1998) and beta tubulin (tub2) using ITS1/ITS4, TEF1/TEF2 and Bt2a/Bt2b (Carbone and Kohn 1999; Glass and Donaldson 1995) were amplified using primer pairs respectively. PCR amplicons were sequenced, and the sequences were deposited in GenBank (accession numbers: ITS: OP880881.1, OP880882.1; tef-1α: OP882579.1, OP882580.1; and tub2: OP882581., OP882582.1). The phylogeny was constructed based on combined ITS, tef-1a, and tub2 regions. Neighbour-Joining (NJ) analysis was conducted and the tree was constructed with the substitution models (branch support was evaluated by 1,000 bootstrap replications). Combined phylogeny confirmed that the sequences shared a common clade with N. clavispora. Hence, morphological, microscopic and molecular characterization confirmed the pathogen as N. clavispora. The pathogenicity test was done on six months old healthy grafts of Priyanka variety (n=9) and repeated thrice. Conidial suspension (2×106 spores/ml) of N. clavispora CLB_SCN1 (15 days old culture) was sprayed on the healthy cashew seedlings, and kept in greenhouse by covering with polythene bags for 24 h (>80 % RH) and maintained under greenhouse condition. The control grafts were inoculated with SDW. The inoculated plants showed blight symptoms after 7-10-day post inoculation and control remained heathy. Re-isolation was done from the symptomatic leaves and identity was confirmed using cultural and molecular studies. Earlier reports showed that, N. clavispora has been reported to cause cardamom leaf blight (Biju et al 2018) and leaf spot disease of plum (Banerjee and Rana 2020). To best of our knowledge, this is the first report of cashew leaf blight disease caused by N. clavispora from India (Farr and Rossman, 2022). Early detection will help farmer in better management and avoiding economic loss caused by N. clavispora.

First report of Colletotrichum siamense causing grape ripe rot in Brazil.

Vitis vinifera cv. BRS Vitória is a seedless black table grape cultivar with an extremely pleasant flavor and is adapted to cultivation in all Brazilian regions. Between November and December 2021, grape berries presenting typical ripe rot symptoms were found in three vineyards located in Petrolina, Pernambuco Brazil. The first symptoms are small and depressed lesions on ripe berries, presenting tiny black acervuli. As the disease progress, lesions enlarge and affect the whole fruit, and abundant orange masses of conidia can be observed. Finally, berries become completely mummified. Symptoms were observed in the three vineyards visited, and disease incidence was above 90%. Some producers are considering eradicating the plantations due to the losses caused by the disease. Control measures used so far are costly and ineffective. Fungal isolation was performed by transferring conidial masses from 10 diseased fruit to plates containing potato dextrose agar medium. Cultures were incubated at 25 °C under continuous light. Seven days after inoculation, three fungal isolates (LM1543-1545) were obtained and subcultivated in pure culture for species identification and pathogenicity test. Isolates presented white to grey cottony mycelia, and hyaline conidia with cylindrical with rounded ends, which resemble the genus Colletotrichum (Sutton 1980). Partial sequences of APN2-MAT/IGS, CAL, and GAPDH loci were amplified, sequenced, and deposited on GenBank (OP643865-OP643872). Isolates from V. vinifera were placed within the clade including the ex-type and representative isolates of C. siamense. The clade was strongly supported (99.8% bootstrap support) in the maximum likelihood multilocus tree of the three loci combined, which confidently assign the isolates to this species. Inoculation on grape bunches was performed to confirm pathogenicity. Grape bunches were surface sterilized for 30 s in 70% ethanol, 1 min in 1.5% NaOCl, rinsed two times with sterile distilled water, and air-dried. Fungal conidial suspensions (106 conidia mL-1) were sprayed to the point of run-off. The negative control was represented by grape bunches sprayed with sterile distilled water. Grape bunches were kept in a humid chamber for 48 h at 25 °C and a light period of 12 h. The experiment was conducted with four replicates (four inoculated bunches per isolate) and repeated once. Typical ripe rot symptoms were observed on grape berries 7 days after inoculation. No symptoms were observed on the negative control. The fungal isolates recovered from inoculated berries were morphologically identical to the C. siamense isolates originally recovered from symptomatic berries collected in the field, fulfilling Koch's postulates. Colletotrichum siamense was reported in association with grape leaves in USA (Weir et al. 2012) and causing grape ripe rot in North America (Cosseboom & Hu 2022). Only C. fructicola, C. kahawae, C. karsti, C. limetticola, C. nymphaeae and C. viniferum were reported causing grape ripe rot in Brazil (Echeverrigaray et al. 2020). To our knowledge, this is the first report of C. siamense causing grape ripe rot in Brazil. This finding is important for disease management because C. siamense has a high phytopathogenic potential due to its wide distribution and host range.

The protective effect of endogenous melatonin on gilthead seabream sperm during cryopreservation

Cryopreservation is widely used for artificial reproduction techniques (ART) but requires good gamete quality to succeed. Considering the endogenously produced melatonin by the fish and its protective role in the organism, the objective of this experiment was to search for the best moment of the day to collect gilthead seabream (Sparus aurata) sperm with better quality and, at the same time, to test the potential protective effect of the exogenous melatonin during sperm freezing and thawing processes. Fish were first sampled every 6 h, corresponding to the beginning of the light period (6 h), mid-light (12h), beginning of the dark period (18 h), and mid-dark (24 h) to determine melatonin concentration in blood plasma by radioimmunoassay (RIA). The analysis showed higher values at 24 h (244 pg/mL) compared with 12 h (99 pg/mL), which allowed the selection of those moments for the next experiment. After, fish were sampled for sperm collection at mid-light (ML, 12 h) and mid-dark (MD, 24 h), and fresh sperm was used to assess gamete quality and for the cryopreservation assay. Sperm samples were cryopreserved to test supplementation of different melatonin concentrations (0.001 mM, 0.01 mM and 0.1 mM) together with a control group without added melatonin. Gamete quality was assessed through spermatozoa concentration and motility (CASA system), cell viability (PI/SYBR-green) and DNA fragmentation (Comet assay). Despite cell viability that was higher at ML, most of the fresh sperm motility parameters did not differ between ML and MD, only linearity (LIN) was enhanced at MD. Nevertheless, in cryopreserved samples, total motility (TM) was significantly higher at MD in all melatonin treatments, control, and fresh samples, revealing an endogenous night-effect. Moreover, spermatozoa concentration was also higher at MD (28.9 × 109/mL) than at ML (20.7 × 109/mL). Supplemented melatonin did not confer extra protection to gilthead seabream sperm during cryopreservation since the tested concentrations did not differ between the control in any sperm quality test. It is here suggested that endogenously produced melatonin may contribute to the improvement of some gamete quality parameters at mid-dark, allowing the aquaculture sector to select better sperm quality in a non-invasive way by choosing it as the best moment of the day for sperm collection.

Tools for photomotor response assay standardization in ecotoxicological studies: Example of exposure to gentamicin in the freshwater planaria Schmidtea mediterranea

Photomotor response assay (PMR) is very useful in an ecotoxicological context because it allows evaluation of behavioral response to potential toxic compounds. However, a lack of procedure standardization makes results comparison difficult between labs and organisms. Here, we aimed to propose five different tools to standardize the PMR procedure so that it may be applied to all model species, regarding: (1) the minimum total sample size, (2) the acclimation period, (3) the number and duration of light and dark phases alternation, (4) the measured behavior, and (5) the statistical analysis. As an example of procedure application, we analyzed the effect of an exposure to the antibiotic gentamicin on the locomotion behavior during PMR in an invertebrate species: the asexual freshwater planaria Schmidtea mediterranea. We encourage future studies using PMR to follow these five tools to improve data analysis and results comparability.

Effects of UHDR and Conventional Irradiation on Behavioral and Cognitive Performance and the Percentage of Ly6G+ CD45+ Cells in the Hippocampus.

We assessed the effects of conventional and ultra-high dose rate (UHDR) electron irradiation on behavioral and cognitive performance one month following exposure and assessed whether these effects were associated with alterations in the number of immune cells in the hippocampus using flow cytometry. Two-month-old female and male C57BL/6J mice received whole-brain conventional or UHDR irradiation. UHDR mice were irradiated with 9 MeV electrons, delivered by the Linac-based/modified beam control. The mice were irradiated or sham-irradiated at Dartmouth, the following week shipped to OHSU, and behaviorally and cognitively tested between 27 and 41 days after exposure. Conventional- and UHDR-irradiated mice showed impaired novel object recognition. During fear learning, conventional- and UHDR-irradiated mice moved less during the inter-stimulus interval (ISI) and UHDR-irradiated mice also moved less during the baseline period (prior to the first tone). In irradiated mice, reduced activity levels were also seen in the home cage: conventional- and UHDR-irradiated mice moved less during the light period and UHDR-irradiated mice moved less during the dark period. Following behavioral and cognitive testing, infiltrating immune cells in the hippocampus were analyzed by flow cytometry. The percentage of Ly6G+ CD45+ cells in the hippocampus was lower in conventional- and UHDR-irradiated than sham-irradiated mice, suggesting that neutrophils might be particularly sensitive to radiation. The percentage of Ly6G+ CD45+ cells in the hippocampus was positively correlated with the time spent exploring the novel object in the object recognition test. Under the experimental conditions used, cognitive injury was comparable in conventional and UHDR mice. However, the percentage of CD45+ CD11b+ Ly6+ and CD45+ CD11b+ Ly6G- cells in the hippocampus cells in the hippocampus was altered in conventional- but not UHDR-irradiated mice and the reduced percentage of Ly6G+ CD45+ cells in the hippocampus might mediate some of the detrimental radiation-induced cognitive effects.

Ammonia impact on the selection of a phototrophic - chemotrophic consortium for polyhydroxyalkanoates production under light-feast / dark-aerated-famine conditions

Phototrophic polyhydroxyalkanoate (PHA) production is an emerging technology for recovering carbon and nutrients from diverse wastewater streams. However, reliable selection methods for the enrichment of PHA accumulating purple phototrophic bacteria (PPB) in phototrophic mixed cultures (PMC) are needed. This research evaluates the impact of ammonia on the selection of a PHA accumulating phototrophic-chemotrophic consortium, towards the enrichment of PHA accumulating PPB. The culture was operated under light-feast/dark-aerated-famine and winter simulated-outdoor conditions (13.2 ± 0.9 °C, transient light, 143.5 W/m2), using real fermented domestic wastewater with molasses as feedstock. Three ammonia supply strategies were assessed: 1) ammonia available only in the light phase, 2) ammonia always present and 3) ammonia available only during the dark-aerated-famine phase. Results showed that the PMC selected under 1) ammonia only in the light and 3) dark-famine ammonia conditions, presented the lowest PHA accumulation capacity during the light period (11.1 % g PHA/g VSS and 10.4 % g PHA/g VSS, respectively). In case 1), the absence of ammonia during the dark-aerated-famine phase did not promote the selection of PHA storing PPB, whereas in case 3) the absence of ammonia during the light period favoured cyanobacteria growth as well as purple sulphur bacteria with increased non-PHA inclusions, resulting in an overall decrease of phototrophic PHA accumulation capacity. The best PHA accumulation performance was obtained with selection under permanent presence of ammonia (case 2), which attained a PHA content of 21.6 % g PHA/g VSS (10.2 Cmmol PHA/L), at a production rate of 0.57 g PHA/L·day, during the light period in the selection reactor. Results in case 2 also showed that feedstock composition impacts the PMC performance, with feedstocks richer in more reduced volatile fatty acids (butyric and valeric acids) decreasing phototrophic performance and leading to acids entering the dark-aerated phase. Nevertheless, the presence of organic carbon in the aerated phase was not detrimental to the system. In fact, it led to the establishment of a phototrophic-chemotrophic consortium that could photosynthetically accumulate a PHA content of 13.2 % g PHA/g VSS (6.7 Cmmol PHA/L) at a production rate of 0.20 g PHA/L·day in the light phase, and was able to further increase that storage up to 18.5 % g PHA/g VSS (11.0 Cmmol PHA/L) at a production rate of 1.35 g PHA/L·day in the dark-aerated period. Furthermore, the light-feast/dark-aerated-famine operation was able to maintain the performance of the selection reactor under winter conditions, unlike non-aerated PMC systems operated under summer conditions, suggesting that night-time aeration coupled with the constant presence of ammonia can contribute to overcoming the seasonal constraints of outdoor operation of PMCs for PHA production.

Attenuation of homeostatic sleep response and rest-activity circadian rhythm in vitamin D deficient mice

ABSTRACT The link between vitamin D deficiency (VDD) and sleep disturbances has long been suggested. However, the direct causality between VDD, sleep disturbances, and circadian rhythm remains unclear. We aimed to characterize sleep-wake behavior and circadian rhythms in an animal model of VDD. VDD was induced by feeding vitamin D-deficient chow, and we analyzed sleep and circadian rhythm parameters. During light period, VDD mice exhibited reduced wake with more frequent wake bouts and increased NREM sleep time. However, during dark period, the wake EEG power spectrum peaked at theta band frequency, and slow-wave energy was suppressed in mice with VDD. Rest-activity analyses revealed increased circadian period, lower wheel counts, and more frequent and short activity bouts during VDD. Combining sleep and circadian data, we found significantly suppressed activities during the hours with a wake duration shorter than 30 minutes. Moreover, mice in VDD state exhibited a negative correlation between wake theta power and hourly wheel-running counts during dark period. Our data point to a direct link between VDD and disturbances in sleep and rest-activity circadian rhythm, featuring frequent wake bouts during the sleeping phase, reduced sleep pressure build-up in dark period, and reduced activity levels due to increased susceptibility to sleepiness.

Contribution of Glutathione Transferases in the Selective and Light-Dependent Effect of Flumioxazin on Winter Wheat (Triticum aestivum L.) and Its Typical Weed Common Poppy (Papaver rhoeas L.)

Glutathione transferases (GSTs) are enzymes that catalyse modifications and conjugations of a range of organic and often cytotoxic compounds. GST enzymes with many functions—such as their conjugation activity against herbicides and their metabolites—can be induced and show light and circadian determination. The enzyme family, which is widespread in its function, also shows great diversity in its structure, which has been linked to its enzyme kinetic characteristics and physiological role at many points. In this study, we aimed to find out the role of different glutathione transferases in the herbicide responses to flumioxazin, as well as to determine how the antioxidant and detoxification response to herbicide treatment changes in the presence and absence of light. One of the herbicide treatments was carried out during the light period in the morning (9:00 a.m.), and the other before the end of the dark period (4:00 a.m.). The decrease in the maximal quantum efficiency of PS II and the reduction in the chlorophyll concentration supported the effect of the herbicide on Papaver rhoeas. In the guaiacol peroxidase POD and GST activity, there were large differences between the cultivated plants and the weed; both enzyme activities were much higher in the case of wheat. According to the activity of the antioxidant defence enzymes and GST gene expression data, the application of the photosynthesis inhibitor herbicide, flumioxazin, in the dark could allow the wheat antioxidant defence to switch on before the herbicide effect could appear in the light period. Phi and tau group GSTs were transcriptionally upregulated by the treatments in wheat plants (especially TaGSTU1B), while fewer changes were detectable in poppy weed (PrGSTU4). Based on our results, in the background of the greater and more successful response to flumioxazin may be—among other things—the higher degree of variability of the GSTU genes of wheat compared to poppies.

Effects of Blindness on Sleep/Wakefulness States in Mice.

To examine the effects of blindness on sleep/wakefulness states, we compared locomotor activity and delayed recovery from isoflurane anesthesia induced by hypnotics during light and dark periods in sighted CBA/N and blind CBA/J mice. Locomotor activity around the switch from the dark to light period significantly differed in both mice. Delayed recovery induced by brotizolam was attenuated in both periods in CBA/J mice. In addition, the period specificity of delayed recovery caused by suvorexant or diphenhydramine in CBA/N mice was abolished in CBA/J mice. These results suggest that blindness impairs sleep quality.

Development and evaluation of an animal health and welfare monitoring system for veterinary supervision of pullet farms.

Regular welfare monitoring throughout rearing of pullets may help to identify problems early and take counteractions timely, which helps in guaranteeing good welfare. The aims of our observational study were (i) to establish and test a welfare monitoring system that can be used during (short) routine veterinary and technical staff visits for pullet flocks, (ii) to use the monitoring system to investigate variability between flocks and (iii) to analyse factors that potentially affect pullets' body weight, uniformity in body weight and mortality. The developed monitoring system tries to minimise the time required while not losing important information. Age-specific recording sheets comprise animal-based indicators of welfare and relevant environmental factors (housing, management, care) to allow for identifying causes of problems and targeted action. Finally, the system was implemented in a cross-sectional study and data collected in 100 flocks (67 organic, 33 conventional) on 28 rearing farms in Austria. Linear mixed models were used to identify factors influencing body weight, uniformity and mortality, both including all flocks (A) and only organic flocks (O) and a linear regression model with all flocks to investigate associations within animal-based indicators. High variability was found between flocks in animal-based indicators. Body weight was higher when the pre-rearing period was shorter (p≤0.001, A&O), with higher intensities of light (p=0.012, O), with only one compared to more stockpersons (p≤0.007, A&O), with a higher number of flock visits per day (p≤0.018, A&O), and a lower avoidance distance (p=0.034, A). Body weight uniformity increased, with age and decreased with the duration of the light period (p=0.046, A), and, amongst others, was higher on organic farms (farming type; p=0.041). The latter may reflect a more uniform level of welfare due to a lower stocking density and lowered effects of social competition. Within organic flocks mortality was lower if pullets had access to a covered veranda (p=0.025) resulting in an overall lower stocking density inside the barn, while in the model including all farms mortality was higher in cases where a disease had been diagnosed. We conclude that our monitoring system can easily be implemented in regular veterinary and technical staff visits, but could also be used by the farmers'. Several easy-to-record animal-based indicators of animal welfare could be analysed more frequently to increase early detection of problems. Implementation of such a routine-based monitoring system with easy-to-assess animal-based parameters and input measures can contribute to better animal health and welfare in pullets.

First report of Tobacco Fusarium Root Rot Caused by Fusarium meridionale in China.

Tobacco (Nicotiana tabacum L.) was an important economic crop in China. A survey in Yunnan Province in the last several years showed that the incidence of tobacco root rot was 3 to 30%. In July 2021, root rot symptoms were observed with an average incidence of 5% on tobacco (cultivar Yunyan 87) in Dali (25.61° N, 100.27° E). Typical disease symptoms included plants stunted at early stages, brown-colored withering lower leaves and roots that became brown. Under high humidity conditions, symptoms of rot expanded in the roots, also the whole plant became wilted and stunted, and some plants ultimately died. Infected pieces of stem tissues and root were dissected and then sterilized with 2% NaOCl for 30 s, rinsed three times with sterile distilled water, and dried with sterilized filter paper. Three pieces were plated onto potato dextrose agar (PDA) for 3 days at 25°C with a 12-h light period. Colonies on PDA were characterized by white to pale yellow flocculent aerial mycelium, and a pink to red pigment in the agar. To induce sporulation, mycelium on PDA was transferred to carnation leaf agar (CLA) medium. After incubation for 7 days, a single spore was isolated from representative isolate 21DL16 for morphological and molecular analyses. Macroconidia observed on CLA were falcate, slightly curved, three to five septate, measured 33.1 to 53.7 × 3.2 to 4.6 μm (n=50), with a typical foot shaped basal cell. Morphological characteristics of the fungus were in agreement with the description of Fusarium graminearum (Leslie and Summerell 2006). For further identification, the internal transcribed spacer (ITS) region rDNA, translation elongation factor 1ɑ (EF-1α) and RNA polymerase II second largest subunit (RPB2) gene were amplified and sequenced using primers ITS1/ITS4 (White et al. 1990), EF1/EF2 (O'Donnell et al. 2015) and RPB2-5F/RPB2-7cR (Reeb et al. 2004), respectively. Although the ITS sequence (GenBank accession no. OM392025) cannot distinguish F. meridionale from F. graminearum, combined phylogenetic analysis of the sequence of TEF1 (ON062055) and RPB2 (ON211932) clearly showed that the pathogen is F. meridionale that the sequences were 100% similarity, 0.0e-value and 100% query coverage to F. meridionale. Pathogenicity studies were conducted on six-leaf-stage tobacco seedlings cultivar Yunyan 87. A conidial suspension (1×105 spores/mL) was poured over the roots of tobacco seedlings. Three seedlings were treated with sterile water that served as controls. All 10 seedlings were maintained at 25°C at 70% relative humidity. After 5 days, the lower leaves showed symptoms of wilting and the roots of all inoculated seedlings become discolored, that were similar with the original symptoms, whereas the control seedlings did not develop symptoms. The fungus reisolated from the inoculated seedlings was identical to F. meridionale using the EF-1α gene sequence. To date, Fusarium root rot on tobacco in China was caused by F. oxysporium (Chen 2013). However, to the best of our knowledge, this is the first report of F. meridionale causing root rot on tobacco in China. Identification of F. meridionale as a root rot agent might provide important insight for disease management practices on tobacco caused by Fusarium species.

Simple procedure to enhance pulsed amperometric detector (PAD) response stability for inulin-type fructans analysis. Application to a case study with chicory taproot

A new electrode management, within the HPAEC-PAD systems, was proposed to measure inulin-type fructans in chicory roots, grown under two lighting periods: 12 h (T-12 h) and 24 h continuous lighting (T-24 h-CL), with the same daily light integral (DLI). The amperometric cell turn-off (PAD-Off) after elution of carbohydrate of interest, allowed the stabilization of the PAD response, avoiding excessive electrode surface oxidation. The enhanced signal stability allowed the application of fucose as internal standard (ISTD) for data normalization, improving the correctness of linear calibration curves and the quantification of fructans in the case study of chicory plants. T-24 h-CL decreased FW and DW of chicory leaves while increasing these parameters in roots. Fructans amount in chicory roots was significantly higher in the T-24-CL photoperiod. The accuracy of prebiotics quantification by PAD-Off emphasized significant differences between light treatments. CL can improve the yield and quality of chicory roots.

Controllable Stress Increases Rapid Eye Movement Sleep in Rats: Regulation by the Central Nucleus of the Amygdala.

Training with inescapable shock (IS; uncontrollable stressor) is followed by significant decreases in rapid eye movement sleep (REM). However, controllability is important in the effects of stress. We examined the effects of escapable shock (ES; controllable stressor) on sleep and whether the central nucleus of the amygdala (CNA) plays a role in regulating these effects. Six Wistar rats implanted with a cannula located in CNA underwent two days of ES training (20 shock presentations; 0.5 mA; 5.0 s maximum duration; 1.0 min interstimulus interval). Five days later, they were re-exposed to the shock context. Following shock training, REM was significantly increased in both light and dark periods. Non-REM (NREM) and total sleep (TS) duration were decreased during the light period. Similar effects on REM and NREM were observed following re-exposure to the training context alone. Microinjections of saline into CNA immediately following ES also produced similar increases in REM, whereas microinjections of muscimol (MUS; GABAA (γ-aminobutyric acid) antagonist) subsequent to ES blocked the increases in REM. These data, along with previous work with ES and IS, demonstrate that stressor controllability is important in determining how stress impacts sleep. Moreover, the results of the microinjection study indicate that the effects of ES on REM are regulated through the CNA.

Does exposure to moonlight affect day/night changes in melatonin and metabolic parameters in Amazonian fish?

Lunar cycle modulates the rhythmic activity patterns of many animals, including fish. The effect of the moonlight cycle on daily melatonin and metabolic parameters was evaluated in matrinxã (Brycon amazonicus) subjected to external natural lighting. Eighty juvenile were distributed in 4 tanks of 1m3 (20 fish/tank) and divided into two groups. One group was exposed to the full moon and the other group to the new moon for 30 days, which corresponds to the duration of the lunar period. At the end of the lunar phase, 6 fish from each group were anesthetized to collect blood, tissue and eye samples at midday and midnight. The comparison between the light and dark periods revealed a significant increase in plasma and ocular melatonin in the last period. However, there was no significant difference for plasma melatonin between moons. Ocular melatonin presented higher concentrations during the new moon. Glucose, total proteins, cortisol, liver glutathione and gill lipid peroxidation were higher in the full moon compared to in the new moon. Plasma triglyceride was higher during the night for the full moon, and the opposite was found for the new moon. Total cholesterol values were higher at night regardless the moon phase. Glutathione in the gills and lipid peroxidation in the liver showed no significant differences. These results highlight the importance of considering both the day and lunar cycles for melatonin and metabolic parameters in species of commercial interest and susceptible to stressful situations in rearing conditions.

Queue Length Estimation Based on Probe Vehicle Data at Signalized Intersections

Queue length is one of the important indexes to evaluate the operation efficiency of signalized intersection and also the key parameter of intersection signal control optimization. Traditional queue length estimation models are mostly based on fixed detection equipment, and the models assumptions are too harsh; there are certain limitations. Based on the probe vehicle data, this paper establishes a model of queue length estimation for signalized intersection based on shockwave theory. First, based on the speed and location data of the probe vehicle, the vehicle density is calculated to estimate the intersection stop line. A real-time calculation method of vehicle arrival rate is proposed to improve the applicability of the model. Then, based on the shockwave theory, the meeting time of the queue forming wave and the queue discharging wave are calculated after the green light is on. Finally, the queue length is summed in sections, including the distance between the last queued probe vehicle and the stop line during the red light period, the length of the subsequent vehicles arriving during the residual red light time, and the newly increased queue length within the queue discharging time. This paper uses the VISSIM software to simulate the actual intersection. The simulation results show that when the penetration of probe vehicle is 50%, 25%, and 10%, their corresponding mean absolute relative error are 11.27%, 27.77%, and 39.12%, respectively. It can be seen that with the increase of penetration, the error gradually decreases. The average absolute relative error is within the acceptable range. After analyzing the existing similar methods, although the accuracy of the method proposed in this paper does not reach the highest level, it has the advantages of simple operation, less computation, and good real-time computation. Relevant research results can provide support for traffic control at signalized intersections.

The effects of intensity, spectral purity and duty cycle on red light-induced hyperopia in tree shrews.

There have recently been several clinical studies suggesting that brief periods of exposure to red light (repeated low-level red light, 'RLRL') may produce a dramatic anti-myopia effect, calling for further investigations into its therapeutic parameters. Unfortunately, many experimental species used in refractive studies develop myopia in response to this wavelength. Tree shrews are the only animal model other than rhesus monkeys that consistently exhibit hyperopic responses to ambient red light. Here, tree shrews were used to study the influence of the spectral purity, duty cycle and intensity of red light on its anti-myopic effect. Juvenile tree shrews (Tupaia belangeri) were raised from 24 to 35 days after eye opening under ambient lighting that was: standard white colony fluorescent light; pure narrow band red light of either 600, 50-100 or 5 lux; red light that was diluted with 10% white light (by lux) or 50% white and 2 s of pure red light that alternated with 2 s of pure white light (50% duty cycle). Refractive measures were taken with a NIDEK ARK-700 autorefractor and axial dimensions with a LenStar LS-900 Axial Biometer. The pro-hyperopia effect of ambient red light was greatly reduced by even small amounts of concurrent white light 'contamination', but remained robust if 2-s periods of pure white light alternated with 2 s of red. Finally, the hyperopic effect of red light was maintained at reduced luminance levels in the 50-100 lux range and only failed at 5 lux. These results have implications for understanding the mechanisms by which ambient red light affects refractive development, and possibly also for clinical therapies using RLRL. Nevertheless, it remains to be determined if the mechanism of the current clinical RLRL therapy is the same as that operating on tree shrews in ambient red light.

Intermediate complex morphophysiological dormancy in seeds of Aconitum barbatum (Ranunculaceae)

BackgroundSeed dormancy and germination are key components of plant regeneration strategies. Aconitum barbatum is a plant commonly found in northeast China. Although it has potential for use in gardening and landscaping, its seed dormancy and regeneration strategy, which adapt to its natural habitat, are not well understood. Our aim was to identify conditions for breaking A. barbatum seed dormancy and determine its dormancy type. Embryo growth and germination were determined by collecting seeds over time in the field. Laboratory experiments that control light, temperature, and stratification period were conducted to assess dormancy breaking and germination, and GA3 was used to identify dormancy type.ResultsSeeds of A. barbatum have undeveloped embryos with physiological dormancy at maturity in autumn. The embryo-to-seed length ratio increases from 0.33 to 0.78 before the emergence of the radical. Under natural environmental conditions, embryo development begins in early winter. Laboratory experiments have shown that long-term incubation under 4 °C (cold stratification) promotes embryo development and seed dormancy break. With an extension of cold stratification, an increase in germination percentages was observed when seeds were transferred from 4 °C to warmer temperatures. Seeds exposed to light during incubation show a higher germination percentage than those kept in the dark. Seed germination can also be enhanced by a 100 mg/L GA3 concentration.ConclusionsSeeds of A. barbatum display intermediate complex morphophysiological dormancy at maturity. In addition to the underdeveloped embryo, there are also physiological barriers that prevent the embryo from germinating. Dormancy breaking of A. barbatum seeds can be achieved by natural winter cold stratification, allowing seeds to germinate and sprout seedlings at the beginning of the following growing season. Our findings provide valuable insights into the seed dormancy and regeneration strategy of A. barbatum, which could facilitate its effective utilization in gardening and landscaping.