Subchronic toxicity of iron-selenium nanoparticles on oxidative stress response, histopathological, and nuclear damage in amphibian larvae Rana saharica.

In this work, we evaluated the subchronic toxicity of FeSe nanoparticles (NPs) in tadpoles of Rana saharica. Tadpoles were exposed for 1-3 weeks to FeSe NPs at 5 mg/L and 100 mg/L rates. Parameters of oxidative stress were measured in whole larvae, and the micronucleus test was performed on circulating blood erythrocytes. We noted a disturbance of the detoxification systems. Enzymatic and non-enzymatic data showed that exposure to FeSe NPs involved a highly significant depletion of GSH, a significant increase in GST activity, and a lipid peroxidation associated with a highly significant increase in MDA. We also noted a neurotoxic effect characterized by a significant inhibition of AChE activity. A micronucleus test showed concentration-dependent DNA damage. This research reveals that these trace elements, in their nanoform, can cause significant neurotoxicity, histopathologic degeneration, cellular and metabolic activity, and genotoxic consequences in Rana larvae.

Bioassay-Guided Isolation of Anti-Candida Biofilm Compounds From Methanol Extracts of the Aerial Parts of Salvia officinalis (Annaba, Algeria).

Salvia officinalis is frequently used in traditional Algerian medicine to treat diverse microbial infections, including oral and vaginal candidiasis. The aerial parts of S. officinalis collected in Annaba, Algeria were extracted in parallel by maceration with four solvents viz. hexane, acetone, methanol and water. All the extracts were tested in vitro against several Candida species: C. albicans, C. glabrata, and C. parapsilosis. Furthermore, the activity against biofilm-forming C. albicans was investigated using bioassay-guided fractionation. A large-scale extract was prepared via maceration in methanol, followed by fractionation on a silica gel column using increasingly polar mixtures of n-hexane, ethyl acetate, methanol, and acetic acid as mobile phase, to yield a total of 150 fractions. Two major active fractions (F-31 and F-39), were further separated by HPLC, resulting in several active chromatographic peaks. Carnosol and 12-methoxy-trans-carnosic acid were isolated as two major active compounds, and identified by a combination of NMR and mass spectrometry. The biofilm inhibitory concentration showed that 12-methoxy-trans-carnosic acid is more effective than carnosol with BIC50 values of 94 µM (95% confidence interval, 78.9-112.1 µM) and 314 µM (95% confidence interval, 200.7-491.2 µM), respectively. The present study supports the traditional use of sage in the treatment of various fungal infections caused by Candida. Further studies of the bioactive compounds in an in vivo Candida biofilm model are required to validate their clinical potential as antifungals.

High diversity of microcystins in a Microcystis bloom from an Algerian lake.

Microcystins (MCs) are cyanobacterial heptapeptides, produced by several genera and species of cyanobacteria, which have been involved in poisoning of animals throughout the world and have also been implicated in human health problems. They are regarded as the most frequently occurring and widespread of the cyanotoxins, with more than 100 MC variants reported to date including the present study. The lake des Oiseaux is a shallow permanent freshwater lake located in north-eastern Algeria. It is an important natural reserve playing a major role for the migratory birds after the crossing of the Mediterranean Sea and from the Sahara desert. In recent years, possibly related to increased eutrophication of the lake, massive blooms of cyanobacteria identified as Microcystis spp. have been observed. A bloom sample collected in September 2013 was analyzed by the serine/threonine phosphatase PP2A inhibition assay and liquid chromatography-mass spectrometry to determine respectively, the total concentration of MCs and the different variants of these toxins present. The results revealed that the Microcystis spp. bloom sample contained microcystins of which 21 putatively congeners were detected. Among these, 12 known microcystins (MC-RR, MC-LR, MC-FR, MC-WR, MC-YR, MC-LA, MC-(H4)YR, MC-HilR, [Asp(3)]MC-RAba, and [Glu(OCH3)(6)]MC-LR) and two new congeners ([Asp(3)]MC-HarAba and [Glu(OCH3)(6)]MC-FR) were characterized, considering their molecular mass and the fragment ions produced by collision-induced dissociation of the [M+H](+) ions. MC-RR was the major (43.4%) in the bloom sample.

Variation in cyanobacterial hepatotoxin (microcystin) content of water samples and two species of fishes collected from a shallow lake in Algeria.

Microcystins (MCs) produced from cyanobacteria can accumulate in freshwater fish tissues. In this study, variations in these toxins content were examined monthly in water samples and two species of fish in Lake Oubeira, Algeria, from April 2010 to March 2011. During the study period, MCs were analyzed using protein phosphatase type 2A (PP2A) inhibition assay. In lake water, total (dissolved and intracellular toxins) MC concentrations by PP2A ranged from 0.028 to 13.4 µg equivalent MC-LR/l, with a peak in September 2010. MC-LR was the dominant variant (90 % of the total) in water samples, followed by MC-YR and MC-(H4)YR. The highest MC concentration in the omnivorous common carp (Cyprinus carpio) was found in the order intestine > hepatopancreas > muscle; however, in the carnivorous European eel (Anguilla anguilla) the order was liver > intestine > muscle. Highest MC concentrations in the intestine tissue of the common carp were found between August and November 2010 where high MC concentrations were detected in water samples, whereas high levels of MCs in the liver of the European eel were found later between January and February 2011. During the entire period of study, the World Health Organization (WHO) lifetime limit for tolerable daily intake was exceeded only in common carp muscle.

First reported case of turtle deaths during a toxic Microcystis spp. bloom in Lake Oubeira, Algeria.

Microcystins analysis was conducted in field cyanobacterial bloom samples and dead terrapin tissues from Lake Oubeira (Algeria) with an aim of studying the cause of the mortality of the freshwater terrapin species Emys orbicularis and Mauremys leprosa during October 2005. The deaths of these two terrapin species were observed during a bloom of Microcystis spp. The total microcystin content per phytoplankton biomass evaluated with the methanol extraction-protein phosphatase methodology was 1.12 mg MCYST-LR equivalents/g dried bloom material. The analysis of this bloom extract by the LC/MS technique demonstrated the presence of three microcystin variants: microcystin-LR (MCYST-LR), microcystin-YR (MCYST-YR), and microcystin-RR (MCYST-RR). Microcystins were also detected in fresh carcasses of terrapin liver, viscera and muscle tissues using the GC/MS after Lemieux oxidation method and the PP2A inhibition assay. The high level of microcystins detected using the Lemieux oxidation-GC/MS method in the liver tissue (1192.8 microg MCYST-LR equivalent/g dw) and in the viscera tissue (37.19 microg MCYST-LR equivalent/g dw) of the species M. leprosa and E. orbicularis, respectively, and the liver crumbling observed after the necropsy examination of the fresh carcass of M. leprosa support the possibility that cyanobacterial microcystins contribute to the turtle mortalities.

A new morphospecies of Microcystis sp. forming bloom in the Cheffia dam (Algeria): seasonal variation of microcystin concentrations in raw water and their removal in a full-scale treatment plant.

Toxic cyanobacterial blooms are an increasing problem in Algeria. The production of cyanotoxins (microcystins) and their presence in drinking water represent growing hazards to human health. In this study, seasonal variations in the concentrations of total microcystins and physicochemical parameters (pH, temperature, dissolved oxygen, nitrate, orthophosphate, and chlorophyll-a) were analyzed in the Cheffia dam (Algeria), mainly used to supply drinking water. The removal of cyanobacterial cells and microcystins was also evaluated in full-scale plant associated with the Cheffia reservoir. The levels of microcystins (MCYSTs) in both raw and drinking water were evaluated using the protein phosphatase type 2A (PP2A) inhibition test as MCYST-LR equivalents. Identification of microcystin variants was achieved by LC/MS/MS. During the period of study (March-December 2004), microscopic observation showed the dominance in the autumn months (September-November) of a new morphospecies of Microcystis sp. The MCYST-LR equivalent concentrations in raw water varied between 50.8 and 28,886 ng L(-1). The highest level of toxins was observed in October 2004 and was significantly correlated with the chlorophyll-a. Three variants of microcystins assigned as microcystin-YR (MCYST-YR), microcystin-LR (MCYST-LR), and 6Z-Adda stereoisomer of MCYST-LR were observed in the crude extract of the Microcystis sp. bloom sample. During the bloom period, total elimination of Microcystis sp. and toxins were achieved through a classical treatment plant comprised of coagulation and flocculation, powdered activated carbon at 15 mg L(-1), slow sand filtration and chlorination before storage.