To meet the increasing global energy demand, expanding exploration for oil and gas reserves as well as associated drilling activities are expected in the Arctic-boreal region where sponge aggregations contribute to up to 90% of benthic biomass. These deep-water sponges along with their microbial endobionts play key roles in the nitrogen cycling in Arctic-boreal ecosystems. This study aimed to investigate the effects of drilling discharges and associated sediment resuspension events on net fluxes of oxygen, ammonium, nitrate and nitrite in three common deep-water sponge species in the form of explants. Sponges were exposed to suspended bentonite and barite, the primary particulate compounds in drilling waste, as well as suspended natural sediment particles for a period of 33 days (on average 10 mg L−1 for 12 h day−1). The exposure period was followed by a pollution abatement period for a further 33 days. No sponge mortality was observed during the experiment. However, exposure to these particles, especially to barite, led to reduced oxygen consumption by up to 33% that was linearly correlated with reduced nitrite/nitrate release by the sponges. The changes in net fluxes were accompanied by decreased tissue oxygenation by up to 54% within the sponges. These findings reveal the effects of fine particles on sponge metabolic processes by reducing aerobic respiration and microbial nitrification, and possibly by favouring anaerobic processes such as microbial denitrification. Most of the sponge responses recovered to their control levels upon the pollution abatement period, but the effects caused by barite may not be reversible. Our findings provide the first insight into the ecological consequences of oil and gas drilling activities on sponge-mediated nitrogen cycling in the Arctic-boreal region.

Kidney vetch (Anthyllis vulneraria L.) is a well-known Zn hyperaccumulator. Zn often occurs with Pb in one ore; thus, plants inhabiting waste dumps are exposed not only to Zn but also to Pb toxicity. While the response of kidney vetch to Zn toxicity is relatively well known, the Pb survival strategy of Anthyllis vulneraria has not been the subject of investigations. The aim of presented research was to determine the survival strategy of kidney vetch exposed to high lead concentrations. Shoot explants of a calamine kidney vetch ecotype were placed on agar media containing 0.0, 0.5, 1.0, and 1.5 mM Pb. Morphological, physiological, and biochemical responses, in particular photosynthetic apparatus of plantlets, were examined. The most pronounced changes were observed in plants grown on media supplemented with 1.5 mM Pb after 8 weeks of culture. Increased dry weight and high lead accumulation were observed in roots. Similarly, in shoots, increased dry weight and a decreased number of newly formed shoots were recorded. The accumulation of lead was many times lower in shoots than in roots. In leaf cells’ ultra-structure, looser arrangement of chloroplast thylakoid grana was observed. Despite the decrease in chlorophyll a and carotenoid content, the photosynthetic apparatus remained efficient due to the lack of photoinhibition and increased electron transport rate beyond photosystem II (PSII). For the first time, an acclimatization mechanism based on maintaining the high efficiency of photosynthetic apparatus resulting from increasing of electron transport rate was described.

Exposure to environmental estrogens and other endocrine-active chemicals can impact reproduction of freshwater fishes. While extensive data exists regarding the effect of estrogens on standard laboratory species, little is known about the sensitivity of freshwater fishes native to North America to these compounds. Current testing strategies for the toxicological assessment of contaminants still rely heavily on studies with live animals, which poses increasing concerns from an economical and ethical perspective. Therefore, the aim of the present study was to investigate the sensitivity of four native species, namely, northern pike (Esox lucius), walleye (Sander vitreus), white sucker (Catostomus commersoni), and juvenile white sturgeon (Acipenser transmontanus), to an environmental estrogen, 17α-ethinylestradiol (EE2), using an in vitro tissue explant approach. Transcript abundances of vitellogenin (VTG) as well as the estrogen receptors (ER) α and β were used as the measuring endpoints as they represent well established biomarkers previously used to assess exposure to estrogens. Transcript abundance of VTG was upregulated in a concentration-dependent manner in each species. Liver explants of male walleye were found to have the greatest sensitivity to EE2, with a lowest observable effect concentration of 300 ng/L (1.0 nM) for VTG transcript abundance, with juvenile white sturgeon having the greatest magnitude of VTG transcript upregulation in exposed tissue (15-fold relative to control). Exposure of liver explants to EE2 resulted in no alteration in transcript abundance of ERβ, whereas upregulation of ERα was observed in northern pike only. Based on in vitro expression of VTG, the species tested were among the species with greatest sensitivity to environmental estrogens tested to date.

In vitro elicitation of an important compound conessine has been done in the bark-derived callus culture of Holarrhena antidysenterica (L.) Wall. employing different elicitors. For induction of callus, green bark explants excised from field-grown plants were cultured on MS medium augmented with different concentrations (0, 1, 2.5, 5, and 10 μM) of various growth regulators such as BA, IBA, NAA, and 2,4-D either alone or in combinations. The maximum amount of conessine (458.18 ± 0.89ᵈ μg/g dry wt.) was achieved in callus developed on MS medium supplemented with 5 μM BA and 5 μM 2,4-D through HPLC analysis. Elicitation in conessine content in the above callus was achieved employing a variety of organic (phenylalanine, tyrosine, chitosan, tryptophan, casein hydrolysate, proline, sucrose, and yeast extract) as well as inorganic elicitors (Pb(NO₃)₂, As₂O₃, CuSO₄, NaCl, and CdCl₂) in different concentrations. The optimum enhancement in conessine content (3518.58 ± 0.28ᵍ μg/g dry wt.) was seen at the highest concentration (200 mg/L) of phenylalanine. The enhancement was elicitor specific and dose dependent. The overall increment of the conessine content was seen in the order of phenylalanine > tryptophan > Pb(NO₃)₂ > sucrose > NaCl > As₂O₃ > casein hydrolysate > CdCl₂ > chitosan > proline > yeast extract > CuSO₄ > tyrosine. The isolation and purification of conessine was done using methanol as a solvent system through column chromatography (CC) and TLC. The isolated compound was characterized by FT-IR, ¹H-NMR, and HRMS which confirmed with the structure of conessine. The bioassays conducted with the isolated compound revealed a strong larvicidal activity against Anopheles stephensi Liston with LC₅₀ and LC₉₀ values being 1.93 and 5.67 ppm, respectively, without harming the nontarget organism, Mesocyclops thermocyclopoides Harada, after 48 h of treatment. This is our first report for the isolation and elicitation of conessine in the callus culture of H. antidysenterica.