Acute exposure to copper induces variable intensity of oxidative stress in goldfish tissues

Copper is an essential element, but at high concentrations, it is toxic for living organisms. The present study investigated the responses of goldfish, Carassius auratus, to 96 h exposure to 30, 300, or 700 μg L^?1 of copper II chloride (Cu²⁺). The content of protein carbonyls was higher in kidney (by 158%) after exposure to 700 mg L^?1 copper, whereas in gills, liver, and brain, we observed lower content of protein carbonyls after exposure to copper compared with control values. Exposure to copper resulted in increased levels of lipid peroxides in gills (76%) and liver (95–110%) after exposure to 300 and 700 μg L^?1 Cu²⁺. Low molecular mass thiols were depleted by 23–40% in liver and by 29–67% in kidney in response to copper treatment and can be used as biomarkers toxicity of copper. The activities of primary antioxidant enzymes, superoxide dismutase and catalase, were increased in liver as a result of Cu²⁺ exposure, whereas in kidney catalase activity was decreased. The activities of glutathione-related enzymes, glutathione peroxidase, glutathione-S-transferase, and glutathione reductase were decreased as a result of copper exposure, but glutathione reductase activity increased by 25–40% in liver. Taken together, these data show that exposure of fish to Cu²⁺ ions results in the development of low/high intensity oxidative stress reflected in enhanced activities of antioxidant and associated enzymes in different goldfish tissues.     

Bioactivity Screening of Antarctic Sponges Reveals Anticancer Activity and Potential Cell Death via Ferroptosis by Mycalols

Sponges are known to produce a series of compounds with bioactivities useful for human health. This study was conducted on four sponges collected in the framework of the XXXIV Italian National Antarctic Research Program (PNRA) in November-December 2018, i.e., Mycale (Oxymycale) acerata, Haliclona (Rhizoniera) dancoi, Hemimycale topsenti, and Hemigellius pilosus. Sponge extracts were fractioned and tested against hepatocellular carcinoma (HepG2), lung carcinoma (A549), and melanoma cells (A2058), in order to screen for antiproliferative or cytotoxic activity. Two different chemical classes of compounds, belonging to mycalols and suberitenones, were identified in the active fractions. Mycalols were the most active compounds, and their mechanism of action was also investigated at the gene and protein levels in HepG2 cells. Of the differentially expressed genes, ULK1 and GALNT5 were the most down-regulated genes, while MAPK8 was one of the most up-regulated genes. These genes were previously associated with ferroptosis, a programmed cell death triggered by iron-dependent lipid peroxidation, confirmed at the protein level by the down-regulation of GPX4, a key regulator of ferroptosis, and the up-regulation of NCOA4, involved in iron homeostasis. These data suggest, for the first time, that mycalols act by triggering ferroptosis in HepG2 cells.     

Apple juice clarification by immobilized pectolytic enzymes in packed or fluidized bed reactors

The catalytic behavior of a mixture of pectic enzymes, covalently immobilized on different supports (glass microspheres, nylon 6/6 pellets, and PAN beads), was analyzed with a pectin aqueous solution that simulates apple juice. The following parameters were investigated: the rate constant at which pectin hydrolysis is conducted, the time (tau(50)) in which the reduction of 50% of the initial viscosity is reached, and the time (tau(comp,dep)) required to obtain complete depectinization. The best catalytic system was proven to be PAN beads, and their pH and temperature behavior were determined. The yields of two bed reactors, packed or fluidized, using the catalytic PAN beads, were compared to the circulation flow rate of real apple juice. The experimental conditions were as follows: pH 4.0, T = 50 degrees C, and beads volume = 20 cm(3). The initial pectin concentration was the one that was present in our apple juice sample. No differences were observed at low circulation rates, while at higher recirculation rates, the time required to obtain complete pectin hydrolysis into the fluidized reactor was found to be 0.25 times smaller than in the packed bed reactor: 131 min for the packed reactors and 41 min for the fluidized reactors.     

Toxicity of the chitin synthesis inhibitors,diflubenzuron and its dichloro-analogue,to Spodoptera littoralis larvae

The activities of the chitin synthesis inhibitors, diflubenzuron and PH 60–38, against Spodoptera littoralis larvae were assayed by feeding treated alfalfa or poisoned wheat bran baits, by allowing the larvae to imbibe sucrose-containing aqueous dispersions of the compounds, and by injection into larvae. PH 60–38 was less active than diflubenzuron. On alfalfa, diflubenzuron had to be fed for at least 2 days to prevent formation of normal pupae and emergence of adults. For very big (480–540 mg) larvae, feeding diflubenzuron at concentrations of 50 mg/litre for 2 days or 2.5 mg/litre for 3 days prevented adult emergence. For 200–250 mg larvae, this was achieved by feeding concentrations of 100 mg/litre for 2 days, 5 mg/litre for 3 days or 3.5 mg/litre for 4 days. In all larvae > 150 mg, mortality in feeding experiments occurred in the prepupal or the pupal stage. Only with 30–50 mg and 100–150 mg larvae was there considerable mortality during moults between larval instars, the larvae being unable to liberate themselves from the old larval skins and head capsules. Diflubenzuron incorporated into wheat bran baits at concentrations of from 2.5 to 10 000 μg/g killed approximately 70–90% of the insects. When imbibed, diflubenzuron was much less toxic as a wettable powder than as a liquid formulation but the two formulations were equitoxic when injected into the larvae.     

Pathologic findings in reintroduced Przewalski's horses (Equus caballus przewalskii) in southwestern Mongolia.

The Przewalski's horse (Equus caballus przewalskii) was extinct in the wild by the mid 1960s. The species has survived because of captive breeding only. The Takhin Tal reintroduction project is run by the International Takhi Group; it is one of two projects reintroducing horses to the wild in Mongolia. In 1997 the first harem group was released. The first foals were successfully raised in the wild in 1999. Currently, 63 Przewalski's horses live in Takhin Tal. Little information exists on causes of mortality before the implementation of a disease-monitoring program in 1998. Since 1999, all dead horses recovered (n = 28) have been examined and samples collected and submitted for further investigation. Equine piroplasmosis, a tick-transmitted disease caused by Babesia caballi or Theileria equi, is endemic in Takhin Tal and was identified as the cause of death of four stallions and one stillborn foal. In December 2000, wolf predation was implicated in the loss of several Przewalski's horses. However, thorough clinical, pathologic, and bacteriologic investigations performed on dead and surviving horses of this group revealed lesions compatible with strangles. The extreme Mongolian winter of 2000-2001 is thought to have most probably weakened the horses, making them more susceptible to opportunistic infection and subsequent wolf predation. Other occasional causes of death since 1999 were trauma, exhaustion, wasting, urolithiasis, pneumonia, abortion, and stillbirth. The pathologic examination of the Przewalski's horses did not result in a definitive diagnosis in each case. Several disease factors were found to be important in the initial phase of the reintroduction, which could potentially jeopardize the establishment of a self-sustaining population.     

The effect of CME 134 onSpodoptera Littoralis Eggs and larvae

CME 134, a new benzoylphenyl urea chitin synthesis inhibitor, was less active than diflubenzuron and BAY SIR 8514, when tested againstSpodoptera littoralis eggs by a dipping method. AgainstS. littoralis larvae the compound was tested by feeding treated alfalfa, topical application and contact with crystalline residues on glass, followed by observation until the adult stage. With both 200–250 and 360–440-mg larvae 100% mortality was obtained by one-day feeding of alfalfa treated with 0.15 ppm a.i. Topical application to 100- and 200-mg larvae showed CME 134 to be about five and nine times more active than BAY SIR 8514 and diflubenzuron, respectively. These differences were even much greater in the contact tests. Cotton field plots were sprayed with either CME 134 or diflubenzuron formulations, leaves were collected at different intervals and fed for one day toS. littoralis larvae in the laboratory. 0.0009% a.i. CME 134 residues gave complete kill of 30–50-mg larvae after 5 and 20 days, and 86% kill after 28 days of aging. With 0.003 and 0.009% a.i., complete kill was obtained in 200–250-mg larvae until 50 days after spraying.     

Role of habitat and landscape in structuring small mammal assemblages in hedgerow networks of contrasted farming landscapes in Brittany,France

In this study, we investigated the environmental factors driving small mammal (rodents and shrews) assemblages in permanent habitat patches in response to a gradient of agricultural intensification. Small mammals were sampled using a trapping standard method in the hedgerow networks of three contrasted landscapes differing by their level of land-use intensity and hedgerow network density (BOC1: slightly intensified; BOC2: moderately intensified and POL: highly intensified). We hypothesized that habitat and landscape characteristics have to be considered to understand the structure of local community. In that way, we carried out a multi-scale study using environmental variables ranging from local habitat (structure and composition of the hedgerows) to hedgerows neighbourhoods in a radius of 300 m (land cover and connectivity around hedges) and to landscape units (three sites). During 1 year, 24 hedgerows were sampled seven times, representing a total of 1,379 captures (86% of rodents and 14% of shrews) and eight species, dominated by the wood mouse (Apodemus sylvaticus) and the bank vole (Clethrionomys glareolus). Inter-site variability was significant and accounted for 18% of total variation in small mammal species abundances. But intra-site variability was also highlighted: species abundance profiles may differ greatly among hedgerows within a site. The more explanatory variables were identified at the different scales of the study: the landscape unit POL was shown to be an important factor in structuring the community, but the predominant factors explaining differences of abundances among hedgerows were about local habitat. In fact, the width of hedges and the tree species richness appeared to be significant and explaining the greatest part of the total variation of the small mammal community composition.     

Efficacy of Neem-Azal S and Margosan-0 against the bean aphid,Aphis fabae Scop.

Margosan-0, a commercial preparation of neem seed kernel extract was tested for its effects as toxicant, deterrent and sterilant against the bean aphidAphis fabae and compared with the German product Neem-Azal S. Second and 4th nymphal instars ofA. fabae, placed on bean plants treated with different concentrations of Margosan-0 or Neem-Azal S, were adversely affected and very high percentage of mortality occurred after 96 hours. In a choice test, higher numbers of adults settled and larviposited on plants treated with Margosan-0 in comparison to the control. Neem-Azal S treated plants showed high deterrence to the adults at high concentrations. The deterrent effect was reflected in a reduced larviposition. Plants treated with different concentrations of either Neem-Azal S or Margosan-0 significantly affected the longevity and fecundity ofA. fabae. The efficacy of Neem-Azal S is more pronounced than that of Margosan-0.     

Tobacco necrosis virus A overcomes local cell death response in Nicotiana tabacum

Tobacco necrosis virus A (TNVA) belongs to the genus Alphanecrovirus in the Tombusviridae family. Symptoms induced by TNVA include necrotic lesions on inoculated leaves in a wide host range. The necrotic cell death response to certain viruses is poorly understood. This study characterizes the mechanisms underlying the local cell death triggered by TNVA in Nicotiana tabacum, showing that it shares several components with the hypersensitive response (HR) mediated by resistance proteins. TNVA perception triggers the accumulation of local viral-derived small interference RNA and the regulation of biological processes related to pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI), including hydrogen peroxide accumulation, cell wall reinforcement, the activation of unfolded protein response and the induction of salicylic acid (SA) and SA-dependent pathways. However, these antiviral defences do not prevent either local virus multiplication or systemic movement, leading to disease development. Nevertheless, SA-deficient NahG tobacco plants challenged with TNVA showed an increase in virus accumulation on noninoculated leaves, which was correlated with the development of systemic necrosis, highlighting the role of SA signalling in TNVA-induced defence. In addition, SA treatment enhanced the local defence response to TNVA infection and suppressed systemic necrosis in N. benthamiana. Taken together, our data suggest that TNVA induces an impaired plant defence response by modulating host factors to persist at low levels in distal tissues.     

Babesiosis in free-ranging chamois (Rupicapra r. rupicapra) from Switzerland

Pathological examination of five adult chamois (Rupicapra r. rupicapra) found dead in two different regions from the Swiss Alps revealed pale mucous membranes and musculature, swollen spleen and haemoglobinuria. Histologically, haemosiderosis in the spleen and centrilobular hepatic necrosis were the predominant findings. On blood smears, small (approximately 0.84-1.47 microm), round to pyriform, peripherally located inclusions were present in the erythrocytes. PCR followed by sequencing of DNA extracted from blood or spleen of the infected animals revealed 99-100% identity of the amplified part of the 18S rRNA gene with GenBank entries attributed to Babesia divergens/Babesia capreoli. This is the first report of fatal Babesia infections in chamois raising the question of an emerging disease in this species.