Screening of Anaesthetics in Adult Zebrafish (Danio rerio) for the Induction of Euthanasia by Overdose

Zebrafish are often euthanized by overdose of anaesthesia. However, fish may have aversion towards some anaesthetics, and protocol efficacy varies between species. Using wild type adult Danio rerio, we assessed time to loss of opercular beat, righting, and startle reflexes during induction of anaesthetic overdose by either tricaine (0.5 g/L or 1 g/L), benzocaine (1 g/L), 2-phenoxyethanol (3 mL/L), clove oil (0.1%), isoeugenol (540 mg/L), lidocaine hydrochloride (1 g/L), or etomidate (50 mg/L). Initial screening demonstrated that benzocaine and buffered lidocaine hydrochloride achieved the fastest loss of reflexes. The rapid induction times were confirmed when retesting using larger batches of fish. The fastest induction was obtained with 1 g/L lidocaine hydrochloride buffered with 2 g/L NaHCO3, in which all adult zebrafish lost reflexes in less than 2 min. Next, we monitored signs of distress during benzocaine or buffered lidocaine hydrochloride overdose induction. The results indicated that buffered lidocaine hydrochloride caused significantly less aversive behaviors than benzocaine. Finally, we tested several buffers to refine the lidocaine hydrochloride immersion. The most efficient buffer for euthanasia induction using 1g/L lidocaine hydrochloride was 2 g/L NaHCO3 with 50 mL/L 96% ethanol, inducing immobility in less than 10 s and with only 2% of adult zebrafish displaying aversive behaviors during treatment.

Evaluación del riesgo ambiental por petróleo crudo en las especies acuáticas Lemna minor, Daphnia magna y Danio rerio

Los derrames de petróleo ocasionan contaminación en los sistemas acuáticos y generan un problema ambiental grave. El objetivo del estudio fue evaluar el riesgo ambiental de los derrames de petróleo crudo mediante bioensayos de toxicidad letal y subletal en Lemna minor, Daphnia magna y Danio rerio. Se determinó la toxicidad de los hidrocarburos totales de petróleo (total petroleum hydrocarbon, TPH) en la fracción acumulada de petróleo mediante bioensayos con un diseño de bloques completos al azar (DBCA) de 6 x 4, con cinco concentraciones, un control y cuatro repeticiones. Las variables analizadas fueron el peso seco y la clorosis (L. minor); la mortalidad y la inmovilidad (D. magna), y la hipoactividad, el escape hacia el fondo y la hipoventilación (D. rerio). Los resultados obtenidos se evaluaron mediante diferencias estadísticas (p<0,05) entre las concentraciones y el control. Se determinaron los valores de la concentración en que no se observa efecto por exposición del contaminante (no observed effect concentration, NOEC) de <0,4 y 3,22 mg/L en las variables de clorosis y peso seco para L. minor. En el caso de D. magna, se obtuvo una concentración efectiva media para la inmovilidad (CE50-48h) de 2,74 mg/L y una concentración letal media (CL50-48h) de 6,22 mg/L. En el ensayo con el pez D. rerio, la variable de hipoactividad dio una NOEC y una concentración más baja en la que se observa efecto por exposición del contaminante (LOEC) de 14,28 y 28,61 mg/L a las 96 h de exposición, respectivamente. Los peces mostraron respuesta de escape hacia el fondo a 3,58 mg/L. En cuanto a la hipoventilación, los valores obtenidos de NOEC y LOEC fueron 7,15 y 14,31 mg/L (96 h). Se determinó el siguiente orden de toxicidad decreciente: L. minor (<0,40 mg/L – clorosis 168 h) > D. magna (1,61 mg/L – inmovilidad 48 h) > D. rerio (<3,58 mg/L – escape hacia el fondo 96 h). Todas las variables analizadas presentaron un cociente de riesgo mayor a 1, lo que dejó en evidenció el riesgo acuático ambiental.

The nature of the manifestation of heterosis is a reliable indicator of a high specific combination ability in a cucumber of the bee-pollinated type

Relevance. One of the most important stages in the selection of agricultural crops for heterosis is the determination of the combinational ability, in particular, the evaluation of the SCS constants of lines of promising hybrid combinations. Purpose: on the basis of the best hybrid combinations, create promising cucumber hybrids for open and protected ground, for various purposes, characterized by a complex of economically useful features, including high fruit yield. Objective: to evaluate the effects of true, hypothetical, competitive heterosis of hybrids and SCA line constants in the studied F1 hybrid combinations.Materials and methods.The research work was carried out in the Pridnestrovian Research Institute of Agriculture in 2014-2018 in greenhouses (spring-summer and summer rotations) and in open ground when growing in a spread and on a trellis. Much attention was paid to the degree and nature of the manifestation of heterosis in F1 hybrids, which were evaluated by early and total yield, the yield of standard fruits, the yield of gherkins and greens. The degree of manifestation of true, hypothetical and competitive heterosis of hybrids is established and the SCA constants in the studied F1 combinations are evaluated. The standards were the F1 Zubrenok hybrids of selection the Pridnestrovian Research Institute of Agriculture and Ajax of the Dutch selection. As testers, JL-6 and L.96 were used. In the studies, selection methods were used-pair and return crosses – backcrosses), self-pollination (incucht), selection (individual, group and mass).Results.With positive effects, all three types of heterosis (true, hypothetical and competitive) were observed in the spring-summer and summer rotations of film greenhouses and open ground when grown in a spread and on a trellis for most economically valuable traits in hybrid combinations 43x57 (seven traits); 71/55x41/86; 65x52 (six); 65x41/86, 65x96, 95x68 (five). High SCA constants were observed in film greenhouses (spring-summer and summer rotations) and in open ground (in the spread and on the trellis) lines in combinations 71/55x41/86 (early, general and standard fruit yield; gherkins yield fractions of 5.1-7.0 and 7.1-9.0 cm (+1.0; +0.9; +10.6; +1.2), (+1.8; +4.9; +39.8; +2.6), (+2.0; +4.4; +36.8; +2.6), (+1.1; +1.5; +12.9; +2.3); 65x52 (gherkins yield fractions of 5.1-7.0 cm) (+12.2; +7.2; +12.5; +7.0); 43x57 (yield of gherkins of the fraction 5.1-7.0 cm) (+1.0; +2.0; +6.7; +2.0).

INFLUENCE OF METALS ON SURVIVAL AND LIFE ACTIVITY OF STARFISHES ASTERIAS RUBENS

The effect of metals chlorides (cobalt, manganese, iron, lead, cadmium and copper) at wide range of concentrations on various aspects of the life of starfishes Asterias rubens has been studied. The experiments included a survival and behavioral responses (righting time) evaluation of starfishes during the seven-day exposure. Copper is the most toxic of the studied metals (half-lethal concentration LC50 = 0,062±0,01 mg/L, 96 hours of exposure), then toxicity decrease in the following order: lead (LC50 = 1,99±0,08 mg/L), cadmium (LC50 = 1,6±0,1 mg/L), cobalt (LC50 = 57,7±2,1 mg/L), manganese (LC50= 79,7±4,3 mg/L). Iron at studied concentrations did not influence the starfish’s survival. Copper, lead and cadmium led to a significant increase of the righting time after 96 hours of exposure. The toxic effects of these metals, except for copper, were revealed at higher concentration then Russian norms for surface waters.

COMPARING THE REPELLENCY OF DIFFERENT PLANT ESSENTIAL OILS AGAINST MOSQUITO CULEX PIPIENS

Due to diminishing man-vector contact, repellency is an important way of preventing vector borne diseases. Mosquitoes control and peoples protection from biting by the mosquitoes are the most significant ways to prevent the diseases that have been transmitted by the mosquitoes. Usage of repellants, especially herbal smokes, is one of the proper methods to protect individuals. In this study, four herbs include Rosa damascena Mill, Lavandula officinalis, Eucalyptus globulus and Mentha longifolia L were examined. After preparing the essential oil of each herb, with a certain concentration, a cotton fabric was stained by using the herbs and at animal test stage, their repelling property against Culex pipiens mosquito was evaluated. Finally, the repelling effect of the fabric stained with the essential oils with the chemical combination repelling N, N-diethyl 3-methyl benzamid, as (DEET) was compared as well. The results of repelling effect of the essential oils of Rosa damascena Mill, Mentha longifolia L, Lavandula officinalis and Eucalyptus globulus was 83%, 96%, 81% and 85%, respectively. Mentha longifolia L (96%) had the highest repelling effect; however Lavandula officinalis (81%) has the least repelling effect respectively. Thus, this work showed that some herbal essential oils can be used as natural repellent materials to improve public health.

Peningkatan dan aktivitas enzim asetilkolinesterase pada nyamuk Aedes aegypti yang diseleksi dengan malation

The Elevated and Activity of Acetilcholinesterase Enzyme on Aedes aegypti Selected by Malathion. The aim of this research was to study the effect of selection by malathion on the activity level Acetilcholinesterase enzyme on Aedes aegypti mosquitoes. Selection of Aedes aegypti larval by mean of malathion have been conducted for 20 generations. During the selection process time has been increased of concentration applied and exposure. For generation 0-5 (F0-F5), a concentration of 25 µl/l (24 ppm) was used to expose the larvae to malathion for five minutes. In generation 6-10 (F6-F10) the concentration has increased to 50 µl/l (48 ppm); in F11-F15 the concentration used was 100 µl/l (96 ppm) whereas in F16-F20 200µl/l (192 ppm) was used. Mosquito generations that would be regarded as representative and reference groups were F0, F5, F10, F15 and F20. The LC50 of F0, F5, F10, F15 and F20 was 0,025; 0,032; 0,042; 0,062 and 0,071 ppm respectively. Increases LT50 values was also observed in Aedes aegypti selected by malathion. The LT50 of F0, F5, F10, F15 and F20 generations was 7,9; 11,3; 18; 30,6 and 33,1 minutes respectively. The low levels of malathion resistance could be conferred by the elevated of α-esterase. The values of the α-esterase in F0, F5, F10, F15 and F20 were 0,155; 0,174; 0,203; 0,209 and 0,215 µmol/min/mg protein respectively. The acetilcholinesterase activities were also raised in F0, F5, F10, F15 and F20, the value of acetilcholinesterase activities were 20,35; 20,26; 23,14; 23,18 and 24,9%.

The influence of metallic ions from CoCr28Mo6 on the osteogenic differentiation and cytokine release of human osteoblasts

Inflammatory reactions associated with osteolysis and implant loosening can be the result of generated CoCr particles and the simultaneous release of ions, which are the consequence of wear at the articulating surfaces of metallic implants. By now, there is little knowledge about the influence of CoCr ions on the viability of human osteoblasts (hOB).Therefore, metallic ions were generated from solid CoCr28Mo6 alloys using electric potentials against hydrogen bridge electrodes. For the cell experiments, hOBs (n ≥ 4) in the third passage were used. Cells were cultured for 24 h before incubated with the generated ion solution in two different concentrations (100 µg/l and 500 µg/l in DMEM with 10 % FCS). Cells incubated without ions served as controls. After 48 h and 96 h cell viability was determined (WST-1 assay, live-dead-stainings). Additionally, gene expression levels of osteogenic markers and cytokines were analysed.A significant increase of metabolic activity and vital cells could be shown for both ion concentrations. Gene expression of alkaline phosphatase and Col1 was highly induced after 96 h for 100 µg/l whereas Col1 protein was significantly reduced on both time points (48 h: p=0.008 (100 µg/l, 500 µg/l); 96 h: p=0,029 (500 µg/l)) compared to the control. Additionally, MMP1 expression was decreased after 48 h and 96 h. IL6 mRNA was concentration-dependent increased after 48 h whereas a significant decrease could be shown for the highest ion concentration (p=0.029). Compared to the lower ion concentration, IL8, MCP1 (p=0.029) and TNF mRNA were clearly reduced for the higher ion solution after 96 h. Our results suggest that ions from CoCr28Mo6 alloys have only marginal effects on osteoblast viability whereas Col1 expression is reduced and cytokine release is induced. In further studies, the effects of metallic ions and particles from CoCr28Mo6 on other cells will be examined.

Fungi obtained on various media from soil under banana trees near Logos in Nigeria

From the soil samples collected from beneath various banana plant, <i>Musa paradisiaca</i> L., 96 different species of soil fungi were isolated on medium: Ohio-Agar, Littmans-Agar, Martins Rose Bengal-Agar and identified. Four species of keratinophilic fungi were isolated by means of To-Ka-Va trap-hair method.

Influence of pegvisomant on serum ghrelin and leptin levels in acromegalic patients

IntroductionPegvisomant (peg) is a GH receptor antagonist. In de novo acromegalic patients with high GH levels, ghrelin and leptin levels are reduced, suggesting a direct GH-mediated effect. The aim of our study was to evaluate whether peg treatment in acromegalic patients may abolish the GH impact on ghrelin and leptin levels.MethodsGhrelin, leptin and endogenous GH were measured in ten peg-treated acromegalic patients (three females/seven males, 47 years (28–57)), ten patients with active (act) and ten patients with inactive disease (inact) as well as in ten gender-, age- and body mass index (BMI)-matched healthy volunteers (controls). Endogenous GH was measured using a special in-house assay without interference by peg; total ghrelin and leptin were determined using a commercial RIA and an immunofluorometric in-house assay respectively.ResultsAge and BMI did not differ significantly between groups. Endogenous GH was significantly higher in peg (6.3 μg/l (1.5–41)) and act (9.3 μg/l (1.7–70)) compared with controls (0.1 μg/l (0.1–3.1)) and inact (0.35 μg/l (0.1–2.0), P<0.001). Ghrelin was significantly higher in peg (232 ng/l (96–351)) compared with act (102 ng/l (33–232), P<0.01), whereas ghrelin was not significantly different between the other groups. Leptin was highest in controls (19 μg/l (4–57)) and lowest in act (6 μg/l (2–21)), but this difference did not reach significance.ConclusionTreatment with peg seems to disrupt the feedback loop of ghrelin and GH, leading to elevated ghrelin levels. Furthermore, peg therapy appears not to have a strong impact on leptin levels, as acromegalic patients with and without peg treatment showed similar leptin levels.

Hyalotekite, (Ba,Pb,K)4(Ca,Y)2Si8(B,Be)2(Si,B)2O28F, a tectosilicate related to scapolite: new structure refinement, phase transitions and a short-range ordered 3b superstructure

Hyalotekite, a framework silicate of composition (Ba,Pb,K)4(Ca,Y)2Si8(B,Be)2 (Si,B)28F, is found in relatively high-temperature (⩾ 500°C) Mn skarns at Långban, Sweden, and peralkaline pegmatites at Dara-i-Pioz, Tajikistan. A new paragenesis at Dara-i-Pioz is pegmatite consisting of the Ba borosilicates leucosphenite and tienshanite, as well as caesium kupletskite, aegirine, pyrochlore, microcline and quartz. Hyalotekite has been partially replaced by barylite and danburite. This hyalotekite contains 1.29–1.78 wt.% Y2O3, equivalent to 0.172–0.238 Y pfu or 8–11% Y on the Ca site; its Pb/(Pb+Ba) ratio ranges 0.36–0.44. Electron microprobe F contents of Långban and Dara-i-Pioz hyalotekite range 1.04–1.45 wt.%, consistent with full occupancy of the F site. A new refinement of the structure factor data used in the original structural determination of a Långban hyalotekite resulted in a structural formula, (Pbl.96Bal.86K0.18)Ca2(B1.76Be0.24)(Sil.56B0.44)Si8O28F, consistent with chemical data and all cations with positive-definite thermal parameters, although with a slight excess of positive charge (+57.14 as opposed to the ideal +57.00). An unusual feature of the hyalotekite framework is that 4 of 28 oxygens are non-bridging; by merging these 4 oxygens into two, the framework topology of scapolite is obtained. The triclinic symmetry of hyalotekite observed at room temperature is obtained from a hypothetical tetragonal parent structure via a sequence of displacive phase transitions. Some of these transitions are associated with cation ordering, either Pb–Ba ordering in the large cation sites, or B–Be and Si–B ordering on tetrahedral sites. Others are largely displacive but affect the coordination of the large cations (Pb, Ba, K, Ca). High-resolution electron microscopy suggests that the undulatory extinction characteristic of hyalotekite is due to a fine mosaic microstructure. This suggests that at least one of these transitions occurs in nature during cooling, and that it is first order with a large volume change. A diffuse superstructure observed by electron diffraction implies the existence of a further stage of short-range cation ordering which probably involves both (Pb,K)–Ba and (BeSi,BB)–BSi.