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Summary: Chlamydia and Chlamydia‐like bacteria are well known to infect several organisms and may cause a wide range of diseases, particularly in ruminants. To gain insight into the prevalence and diversity of these intracellular bacteria, we applied a pan‐Chlamydiales real‐time PCR to 1,134 veterinary samples taken from 130 Tunisian ruminant herds. The true adjusted animal population‐level prevalence was 12.9% in cattle, against 8.7% in sheep. In addition, the true adjusted herd‐level prevalence of Chlamydiae was 80% in cattle and 25.5% in sheep. Chlamydiales from three family‐level lineages were detected indicating a high biodiversity of Chlamydiales in ruminant herds. Our results showed that Parachlamydia acanthamoebae could be responsible for bovine and ovine chlamydiosis in central‐eastern Tunisia. Multivariable logistic regression analysis at the animal population level indicated that strata and digestive disorders variables were the important risk factors of bovine and ovine chlamydiosis. However, origin and age variables were found to be associated with bovine and ovine chlamydiosis, respectively. At the herd level, risk factors for Chlamydia positivity were as follows: abortion and herd size for cattle against breeding system, cleaning frequency, quarantine, use of disinfectant and floor type for sheep. Paying attention to these risk factors will help improvement of control programs against this harmful zoonotic disease. [ABSTRACT FROM AUTHOR]

A coagulation–flocculation process was used to treat water-based printing ink wastewater in optimal conditions. A biological process treatment was combined to coagulation–flocculation process using fungal biomass biosorption capacity. Combined process has a significant effect on color removal and on COD removal (80%). The potential of reusing treated wastewater in agriculture was investigated. Germination tests of five species were carried out using treated and untreated wastewater and show that these effluents could be adequately recycled in culture irrigation. [ABSTRACT FROM AUTHOR]

Abstract Background The health benefits of omega-3 and omega-6 polyunsaturated fatty acids (PUFA) are generally recognized. Unfortunately, in most Mediterranean countries, the recommended daily intake of these compounds is rarely met. Therefore, enrichment of commonly occurring foods can boost intake of these fatty acids. In this regard, eggs are an interesting target, as they form an integral part of the diet. Result Zeolite (Clinoptilolites) was added to Laying Hens feed at concentrations 1% or 2% and was evaluated for its effects on performance of the production and on egg quality. The Laying Hens were given access to 110 g of feed mixtures daily that was either a basal diet or a ‘zeolite diet’ (the basal diet supplemented with clinoptilolite at a level of 1% or 2%). It was found that zeolite treatment had a positive and significatif (p Conclusion This study showed the significance of using zeolite, as a feed additive for Laying Hens, as part of a comprehensive program to control egg quality and to increase level of polyunsaturated fatty acids on egg.

Abstract Background Increasing consumer demand for healthier food products has led to the development of governmental policies regarding health claims in many developed countries. In this context, contamination of poultry by food-borne pathogens is considered one of the major problems facing the progress of the poultry industry in Tunisia. Result Zeolite (Clinoptilolites) was added to chicken feed at concentrations 0,5% or 1% and was evaluated for its effectiveness to reduce total flora in chickens and its effects on performance of the production. The broilers were given free and continuous access to a nutritionally non-limiting diet (in meal form)that was either a basal diet or a' zeolite diet' (the basal diet supplemented with clinoptilolite at a level of 0,5% or 1%). It was found that adding zeolite in the broiler diet significantly (p < 0,05) reduced total flora levels, as compared to the control, on the chicken body. In addition, it was found that zeolite treatment had a positive effect on performance production and organoleptic parameters that were measured and mainly on the increase level of Omega 3 fatty acid. Conclusion This study showed the significance of using zeolite, as a feed additive for broilers, as part of a comprehensive program to control total flora at the broiler farm and to increase level of Omega 3 fatty acid on the chicken body.

Microalgae are capable of generating numerous metabolites that possess notable biological activities and hold substantial promise for various industrial applications. Nevertheless, the taxonomic diversity of these photosynthetic microorganisms has not received thorough investigation. Using the 18S rRNA encoding gene, a recently discovered strain originating from the Tunisian coast (the governorate of Mahdia) was identified as a member of the Porphyridium genus. The growth response as well as the metabolite accumulation of Porphyridium sp. to different culture media (Pm, F/2, and Hemerick) was investigated over a period of 52 days. The highest biomass production was recorded with Pm medium (2 × 10 7 cell/mL). The apparent growth rates (µ) and the doubling time (D t ) were about 0.081 day -1 and 12.34 days, respectively. The highest chlorophyll a (0.678 ± 0.005 pg/cell), total carotenoids (0.18 ± 0.003 pg/cell), phycoerythrin (3.88 ± 0.003 pg/cell), and proteins (14.58 ± 0.35 pg/cell) contents were observed with F/2 medium. Cultivating Porphyridium sp. in both F/2 and Hemerick media yielded similar levels of starch accumulation. The Hemerick medium has proven to be the most suitable for the production of lipids (2.23% DW) and exopolysaccharides (5.41 ± 0.56 pg/cell).

The olive oil industry generates 30 million cubic meters of olive mill wastewaters (OMWWs) annually. OMWWs are a major environmental concern in the Mediterranean region due to their high organic matter content, suspended solids, unpleasant odor, and dark color. The application of primary treatments such as coagulation-flocculation, adsorption, and hybrid systems combining coagulation-flocculation with adsorption has enabled to remove part of the organic matter, color, turbidity, and growth-inhibiting compounds from OMWWs. Among these methods, the hybrid system combining activated carbon and chitosan has proven to be the best removal efficiency. Subsequently, secondary treatment involving the cultivation of Chlorella sp. on OMWWs pretreated with chitosan achieved the highest maximal specific growth rate (0.513 ± 0.022 day⁻ 1 ) and biomass productivity (0.621 ± 0.021 g/L/day). Notably, the fatty acids (FA) profile produced by Chlorella sp. cells grown under these conditions differed, underscoring the potential of OMWWs as a microalgal growth medium. This innovative approach not only addresses environmental issues but also opens new avenues for sustainable bioproducts., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Microalgae have gained recognition as versatile candidates for the remediation of heavy metals (HMs). This study investigated the biosorption potential of Dunaliella sp. AL1 for copper (Cu(II)) and hexavalent chromium (Cr(VI)) in aqueous solutions. The marine microalga Dunaliella sp. AL1 was exposed to half-sublethal concentrations of both metals in single and bimetallic systems, and responses in algal growth, oxidative stress, photosynthetic pigment production, and photosynthetic performance were evaluated. Cu and/or Cr exposure increased the generation of reactive oxygen species (ROS) in microalgae cells but did not impact algal growth. In terms of photosynthesis, there was a decrease in chlorophylls and carotenoids production in the microalgae culture treated with Cr, either alone or in combination with Cu. The study recorded promising metal removal efficiencies: 26.67%-20.11% for Cu and 94.99%-95.51% for Cr, in single and bimetallic systems, respectively. FTIR analysis revealed an affinity of Cu and Cr ions towards aliphatic/aldehyde C-H, N-H bending, and phosphate groups, suggesting the formation of complex bonds. Biochemical analysis of microalgae biomass collected after the removal of Cr alone or in combination with Cu showed a significant decrease in total carbohydrate content and soluble protein levels. Meanwhile, higher lipid accumulation was recorded and evidenced by BODIPY 505/515 staining. Fatty acid composition analysis by GC revealed a modulation in lipid composition, with a decrease in the ratio of unsaturated fatty acids (UFA) to saturated fatty acids (SFA), in response to Cu, Cr, and Cu-Cr exposure, indicating the suitability of the biomass for sustainable biofuel production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

With increasing demand of fossil fuels and water pollution and their environmental impacts, marine green microalgae have gained special attention in both scientific and industrial  fields. This is due to their fast growth in non-arable lands with high photosynthetic activity, their metabolic plasticity, as well as their high CO 2 capture capacity. Tetraselmis species, green and eukaryotic microalgae, are not only considered as a valuable source of biomolecules including pigments, lipids, and starch but also widely used in biotechnological applications. Tetraselmis cultivation for high-value biomolecules and industrial use was demonstrated to be a non-cost-effective strategy because of its low demand in nutrients, such as phosphorus and nitrogen. Recently, phycoremediation of wastewater rich in nutrients, chemicals, and heavy metals has become an efficient and economic-alternative that allows the detoxification of waters and induces mechanisms in algal cells for biomolecules rich-energy synthesis to regulate their metabolic pathways. This review aims to shed light on Tetraselmis species for their different culture conditions and metabolites bioaccumulation, as well as their human health and environmental applications. Additionally, phycoremediation of contaminants associated to biofuel production in Tetraselmis cells and their different intracellular and extracellular mechanisms have also been investigated., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A polysaccharide extracted from the brown alga Cystoseira compressa (CCPS) was evaluated as a food additive to extend the shelf-life of raw beef meat. The antioxidant potential of CCPS was demonstrated by its inhibition of β-carotene bleaching (64.28 %), superoxide radicals (70.12 %), and hydroxyl radicals (93 %) at a concentration of 10 mg/ml. The polysaccharide also showed antibacterial activity with MIC values between 6.25 mg/ml and 50 mg/ml against five foodborne pathogenic bacteria. Furthermore, CCPS exhibited excellent functional, foaming, and emulsifying properties. Furthermore, microbiological and chemical effects of CCPS at concentrations equivalent to 1 MIC (CCPS-1), 2 MIC (CCPS-2), and 4 MIC (CCPS-3) were conducted. Chemical analyses showed that treated beef had significantly reduced TBARS levels below 2 mg MDA/kg at day 14. The treatment also decreased carbonyl groups, improved heme iron transformation, inhibited microbial growth (p < 0.05), and kept MetMb levels below 40 % by day 14. Moreover, two multivariate approaches, principal component analysis (PCA) and hierarchical cluster analysis (HCA), were effectively used to analyze the results characterizing the main attributes of the stored meat samples. In conclusion, these findings demonstrated that CCPS could be employed as a functional and bioactive component in the meat industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

ABSTRACT Algae constitute a significant part of marine biodiversity. They represent a renewable source of bioactive metabolites from drug development and therapeutic fields. Fucoxanthin and β -carotene from the brown macroalgae Halopteris scoparia, were extracted using conventional organic solvent extraction, then purified, to homogeneity, based on various chromatographic principles. Their effects on digestive enzymes and harmful bacteria were investigated. The capacities of both purified pigments to inhibit α -amylase and trypsin enzymes were evaluated. Purified fucoxanthin and β -carotene exhibited interesting α -amylase inhibition activities, with IC 50 of 300 and 500 µg/mL, respectively. Moreover, trypsin inhibition activities were detected using purified these two pigments. The antibacterial potential of the purified pigments was evaluated. β -carotene showed to be a great antibacterial natural compound against gram-positive and gram-negative bacteria such as Listeria monocytogenes , Staphylococcus aureus and Salmonella enterica with Minimal Inhibitory Concentration (MIC) of about 0.225, 0.1125, 0.225 µg/mL, respectively. Those findings are in favor of the exploitation of H. scoparia pigments in therapeutic fields as an antidiabetic source directly by the inhibition of α -amylase and trypsin as well as antibacterial agents against gastrointestinal infections.

Microbial extracellular polymeric substances (EPS) have recently emerged as significant contributors in diverse biotechnological applications. Extracellular polymeric substances (EPS), produced by a Navicula salinicola strain, have been studied for potential applications in a specific heavy metal (lead (Pb II)) removal from wastewater. The optimisation of operational parameters, mainly pH, Pb and EPS concentrations, using the Box-Behnken design (BBD) was undertaken to enhance lead uptake. The higher Pb adsorption capacity reached 2211.029 mg/g. Hydroxyl, carbonyl, carboxyl, phosphoric, and sulfhydryl groups were identified quantitatively as potential sites for Pb adsorption. EPS exhibited a notable flocculation rate of 70.20% in kaolin clay at a concentration of 15 mg/L. They demonstrated an emulsifying activity greater than 88%, showcasing their versatile potential for both sedimentation processes and stabilising liquid-liquid systems. EPS could be excellent nonconventional renewable biopolymers for treating water and wastewater.

Soil contamination by heavy metals is one of the major problems that adversely decrease plant growth and biomass production. Inoculation with the plant growth-promoting rhizobacteria (PGPR) can attenuate the toxicity of heavy metals and enhancing the plant growth. In this study, we evaluated the potential of a novel extremotolerant strain (IS-2 T) isolated from date palm rhizosphere to improve barley seedling growth under heavy metal stress. The species-level identification was carried out using morphological and biochemical methods combined with whole genome sequencing. The bacterial strain was then used in vitro for inoculating Hordeum vulgare L. exposed to three different Cr, Zn, and Ni concentrations (0.5, 1, and 2 mM) in petri dishes and different morphological parameters were assessed. The strain was identified as Bacillus glycinifermentans species. This strain showed high tolerance to pH (6-11), salt stress (0.2-2 M), and heavy metals. Indeed, the minimum inhibitory concentrations at which bacterium was unable to grow were 4 mM for nickel, 3 mM for zinc, more than 8 mM for copper, and 40 mM for chromium, respectively. It was observed that inoculation of Hordeum vulgare L. under metal stress conditions with Bacillus glycinifermentans IS-2 T stain improved considerably the growth parameters. The capacity of the IS-2 T strain to withstand a range of abiotic stresses and improve barley seedling development under lab conditions makes it a promising candidate for use as a PGPR in zinc, nickel, copper, and chromium bioremediation., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Marine algae are the most abundant resource in the marine environment and are still a promising source of bioactive compounds including hydrocolloids. This study contributes to the evaluation of the biological and biotechnological potentials of two water soluble polysaccharides, namely alginates (AHS) and fucoidan (FHS), extracted and purified from Halopteris scoparia, an abundant Tunisian brown macroalgae collected in Tunisia (Tabarka region). The total sugars, neutral monosaccharides, uronic acids, proteins, polyphenols, and sulfate groups contents were quantified for both fractions, as well as their functional groups and primary structural features by Fourier transform infrared spectroscopy, ionic and/or gas chromatography and nuclear magnetic resonance analyses. AHS and FHS showed significant anti-inflammatory (IC 50  ≈ 1 mg/mL), anticoagulant (e.g., 27-61.7 for the activated partial thromboplastin time), antihyperglycemic (0.1-40 μg/mL) and anti-trypsin (IC 50  ≈ 0.3-0.4 mg/mL) effects. FHS and a hydrolyzed fraction showed a very promising potential against herpes viruses (HSV-1) (IC 50  < 28 μg/mL). Besides, AHS and two hydrolyzed fractions were able to stimulate the natural defenses of tomato seedlings, assessing their elicitor capacity, by increasing the activity of phenylalanine ammonia-lyase (66-422 %) but also significantly changing the polyphenol content in the leaves (121-243 %) and roots (30-104 %) of tomato plants., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Guillaume Pierre reports financial support was provided by Campus France. Slim Abdelkafi reports financial support was provided by Tunisian Ministry of Higher Education and Scientific Research., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Mercury (Hg) is a global pollutant that affects numerous marine aquatic ecosystems. We isolated Chlorococcum dorsiventrale Ch-UB5 microalga from coastal areas of Tunisia suffering from metal pollution and analyzed its tolerance to Hg. This strain accumulated substantial amounts of Hg and was able to remove up to 95% of added metal after 24 and 72 h in axenic cultures. Mercury led to lesser biomass growth, higher cell aggregation, significant inhibition of photochemical activity, and appearance of oxidative stress and altered redox enzymatic activities, with proliferation of starch granules and neutral lipids vesicles. Such changes matched the biomolecular profile observed using Fourier Transformed Infrared spectroscopy, with remarkable spectral changes corresponding to lipids, proteins and carbohydrates. C. dorsiventrale accumulated the chloroplastic heat shock protein HSP70B and the autophagy-related ATG8 protein, probably to counteract the toxic effects of Hg. However, long-term treatments (72 h) usually resulted in poorer physiological and metabolic responses, associated with acute stress. C. dorsiventrale has potential use for Hg phycoremediation in marine ecosystems, with the ability to accumulating energetic reserves that could be used for biofuel production, supporting the notion of using of C. dorsiventrale for sustainable green chemistry in parallel to metal removal., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Alginates extracted from two Moroccan brown seaweeds and their derivatives were investigated for their ability to induce phenolic metabolism in the roots and leaves of tomato seedlings. Sodium alginates (ALSM and ALCM) were extracted from the brown seaweeds Sargassum muticum and Cystoseira myriophylloides , respectively. Low-molecular-weight alginates (OASM and OACM) were obtained after radical hydrolysis of the native alginates. Elicitation was carried out by foliar spraying 20 mL of aqueous solutions (1 g/L) on 45-day-old tomato seedlings. Elicitor capacities were evaluated by monitoring phenylalanine ammonia-lyase (PAL) activity, polyphenols, and lignin production in the roots and leaves after 0, 12, 24, 48, and 72 h of treatment. The molecular weights (M w ) of the different fractions were 202 kDa for ALSM, 76 kDa for ALCM, 19 kDa for OACM, and 3 kDa for OASM. FTIR analysis revealed that the structures of OACM and OASM did not change after oxidative degradation of the native alginates. These molecules showed their differential capacity to induce natural defenses in tomato seedlings by increasing PAL activity and through the accumulation of polyphenol and lignin content in the leaves and roots. The oxidative alginates (OASM and OACM) exhibited an effective induction of the key enzyme of phenolic metabolism (PAL) compared to the alginate polymers (ALSM and ALCM). These results suggest that low-molecular-weight alginates may be good candidates for stimulating the natural defenses of plants.

Parachlamydia acanthamoebae and Simkania negevensis, two Chlamydia-like bacteria, have been recently recognized as emerging human respiratory pathogens. The prevalence and frequency of these bacteria in the environment and among atypical pneumonia patients are still underestimated by classical cultures, immunohistochemistry and serology which are non-specific, long and tedious methods. This study aims to develop a new duplex probe-based q-PCR assay for the simultaneous detection and quantification of P. acanthamoebae and S. negevensis. The selected hydrolysis probes displayed no cross-reaction with the closely related Chlamydia or the other tested waterborne pathogens. The assay achieved a large dynamic range for quantification (from 5 × 10 6 to 5 DNA copies/reaction). Efficiencies of FAM and JOE label probes weren't affected when they were combined. They were close to 100%, indicating the linear amplification. The application of this diagnostic tool resulted in 9/47 (19%) and 4/47 (8.5%) positive water samples for P. acanthamoebae and S. negevensis, respectively. P. acanthamoebae was also covered from 2/78 (2.5%) respiratory specimens and only one case (1/200 = 0.5%) of P. acanthamoebae and SARS-CoV-2 co-infection was noticed. While S. negevensis wasn't detected in clinical samples, the developed duplex q-PCR was shown to be an accurate, highly sensitive, and robust diagnostic tool for the detection and quantification of P. acanthamoebae and S. negevensis., Competing Interests: Declaration of competing interest No conflict of interest is declared., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Human respiratory infections caused by a large variety of microbial pathogens are the most common diseases responsible for hospitalization, morbidity and mortality. Parachlamydia acanthamoebae, a Chlamydia-related bacterium, has been found to be potentially associated with these diseases. An early and accurate diagnosis of this pathogen could be useful to avoid the potential respiratory complications linked especially to COVID-19 patients and to set suitable outbreak control measures. A TaqMan-PCR assay was developed to detect and quantify Parachlamydia acanthamoebae in environmental and clinical samples from patients of all ages with COVID-19. The selected hydrolysis probe displayed no cross-reaction with the closely related Chlamydia or the other tested pathogens. This q-PCR achieved good reproducibility and repeatability with a detection limit of about 5 DNA copies per reaction. Using this q-PCR assay, Parachlamydia acanthamoebae was detected in 2/78 respiratory specimens and 9/47 water samples. Only one case (1.3%) of Parachlamydia acanthamoebae and SARS-COV-2 co-infection was noticed. To our knowledge, the combination of these two respiratory pathogens has not been described yet. This new TaqMan-PCR assay represents an efficient diagnostic tool to survey Parachlamydia acanthamoebae on a large-scale screening programs and also during outbreaks., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Karlodinium veneficum is a toxic benthic globally distributed dinoflagellate which has direct impacts on human health and the environment. Early and accurate detection of this harmful algal bloom-forming species could be useful for potential risks monitoring and management. In the present work, a real-time PCR targeting the internal transcribed spacer ribosomal DNA region for the specific detection and absolute quantification of K. veneficum was designed. Then, the assay conditions were adjusted and validated. The developed qPCR was highly specific for the target species and displayed no cross-reactivity with closely related dinoflagellates and/or other microalgal species commonly distributed along the Tunisian coast. Its lowest detection limit was 5 rDNA copies per reaction, which is often considered satisfying. qPCR assay enumeration accuracy was evaluated using artificially inoculated environmental samples. The comparison of the cell abundance estimates obtained by qPCR assay with the theoretical estimates showed no statistically significant difference across a range of concentrations. We suggest that the qPCR approach developed in the present study may be a valuable tool to investigate the distribution and seasonal dynamics of K. veneficum in marine environments., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Due to the increase in industrial demand for new biosourced molecules (notably bioactive exopolysaccharides (EPS)), microalgae are gaining popularity because of their nutraceutical potential and benefits health. Such health effects are delivered by specific secondary metabolites, e.g., pigments, exopolysaccharides, polyunsaturated fatty acids, proteins, and glycolipids. These are suitable for the subsequent uses in cosmetic, nutraceutical, pharmaceutical, biofuels, biological waste treatment, animal feed and food fields. In this regard, a special focus has been given in this review to describe the various methods used for extraction and purification of polysaccharides. The second part of the review provides an up-to-date and comprehensive summary of parameters affecting the microalgae growth and insights to maximize the metabolic output by understanding the intricacies of algal development and polysaccharides production. In the ultimate part, the health and nutraceutical claims associated with marine algal bioactive polysaccharides, explaining their noticeable potential for biotechnological applications, are summarized and comprehensively discussed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

The development of eco-friendly cosmetic such as those from microalgae for skin regeneration and collagen synthesis has gained a great interest worldwide. Accordingly, the potential of microalgae biomass as source of anti-aging cosmetic cream with high antioxidant activity has been investigated. Stabilities and sensory characteristics of cosmetic creams supplemented with Spirulina, Tetraselmis sp. and Dunaliella sp. at 0.5, 1.5 and 2.5%, respectively, revealed a conservation of physico-chemical and preliminary stability properties of formulations. To analyze physico-chemical and textural parameters, accelerated stability study was evaluated under two thermal conditions (25 and 40 °C) during 90 days. Results showed that pH values of all formulations were within the limits of normal skin pH range under storage time at 25 and 40 °C. During this period, the colored creams showed a significant changes of a* and b* indices, reflecting the instability of microalgae colors. Microalgae modified the textural characteristics of emulsions. The Tetraselmis sp. containing-cream had the lowest (P < 0.05) values of hardness, springiness, and cohesiveness. The 0.5% Spirulina containing-cream had the best stable consistency and adhesiveness under time and temperature variations. It exhibited the best properties to be used for skin care products. Thanks to their high content in bioactive macromolecules, microalgae considerably improved the antioxidant activity of the new formulated skin creams., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Gymnodinium catenatum is a dinoflagellate known to cause paralytic shellfish poisoning (PSP), commonly associated with human muscular paralysis, neurological symptoms, and, in extreme cases, death. In the present work, we developed a real-time PCR-based assay for the rapid detection of the toxic microalgal species, G. catenatum, in environmental bivalve mollusc samples as well as seawater samples. G. catenatum-specific primers and probe were designed on the ITS1-5.8S-ITS2 rDNA region. Hydrolysis probe qPCR assay was optimized. ITS1-5.8S-ITS2 rDNA region copy numbers per G. catenatum cell genome were estimated to be 122.73 ± 5.54 copies/cell, allowing cell quantification. The application of the optimized qPCR assay for G. catenatum detection and quantification in field samples has been conducted, revealing high sensitivity (detection of around 1.310 5 cells/L of seawater samples. Thus, the designed hydrolysis probe qPCR assay could be considered an efficient tool for phytoplankton monitoring whilst ensuring accuracy and sensitivity and providing cost and time savings., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Given the increasing interest that is being paid to polysaccharides derived from algae as plant natural defense stimulators, the degree of sulfation of exopolysaccharides produced by P. sordidum for inducing defense responses in date palm vitroplants was investigated. Firstly, the culture parameters of P. sordidum were optimized to maximize the amount of sulfate in EPS using a Box-Behnken experimental design and the elicitor effects of two EPS which differ in the sulfation degrees were compared. Results demonstrated that the concentrations of NaCl, NaNO 3 and MgSO 4 set at 28, 0.54 and 16.31 g/L, respectively yielded the best sulfate contents. To elucidate defense-inducing activities in date palm vitroplants, EPS with the highest sulfate content (EPS1) were prepared for comparison with those obtained under standard conditions (EPS0). A fucoidan extracted from Cystoseira compressa was used as positive control and MgSO 4 as negative control. Both EPS and the fucoidan displayed H 2 O 2 accumulation and expression of PR1, SOD, PAL and WRKY genes. Interestingly, EPS1 was significantly more bioactive than EPS0 and the fucoidan suggesting that the elicitor activity is positively correlated with the sulfate groups content of this polysaccharide., (Copyright © 2022 Elsevier Masson SAS. All rights reserved.)

Phycoremediation of heavy metals from contaminated waters by oleaginous microalgae is an eco-friendly and emerging trend. Different concentrations of toxic metals such as nickel (Ni), chromium (Cr) and cobalt (Co) were added in Tetraselmis sp. culture media. Mixture Design was used to model the effect of these metals on cell growth, lipid production and heavy metals removal. Tetraselmis sp. was identified as an outstanding Ni, Cr and Co accumulator with bioconcentration factors of 675.17, 584.9 and 169.81 within binary mixtures (Ni × Cr), (Cr × Co) and (Cr × Co) at 6 mg/L, respectively. Optimization studies showed that the highest cell growth (9.22 × 10 5 cells/mL), lipid content (31% Dry Weight) and metals removal (91%) were obtained with the optimum binary mixture Ni (54.45 %) and Cr (45.45 %). This work presents interesting results revealing the potential of Tetraselmis sp. for nickel removal up to 97 % combined to its potential for biodiesel production., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

A novel sulfated xylogalactan (JASX) was extracted and purified from the rhodophyceae Jania adhaerens . JASX was characterized by chromatography (GC/MS-EI and SEC/MALLS) and spectroscopy (ATR-FTIR and 1 H/ 13 C NMR) techniques. Results showed that JASX was constituted by repeating units of (→3)-β-d-Gal p -(1,4)-3,6-α-l-AnGal p -(1→) n and (→3)-β-d-Gal p -(1,4)-α-l-Gal p -(1→) n substituted on O -2 and O -3 of the α-(1,4)-l-Gal p units by methoxy and/or sulfate groups but also on O -6 of the β-(1,3)-d-Gal p mainly by β-xylosyl side chains and less by methoxy and/or sulfate groups. The M w , M n , Đ, [ η ] and C* of JASX were respectively 600 and 160 kDa, 3.7, 102 mL.g -1 and 7.0 g.L -1 . JASX exhibited pseudoplastic behavior influenced by temperature and monovalent salts and highly correlated to the power-law model and the Arrhenius relationship. JASX presented thixotropic characteristics, a gel-like viscoelastic behavior and a great viscoelasticity character. JASX showed important antioxidant activities, outlining its potential as a natural additive to produce functional foods.

The coronavirus pandemic (COVID-19) has created an urgent need to develop effective strategies for prevention and treatment. In this context, therapies against protease M pro , a conserved viral target, would be essential to contain the spread of the virus and reduce mortality. Using combined techniques of structure modelling, in silico docking and pharmacokinetics prediction, many compounds from algae were tested for their ability to inhibit the SARS-CoV-2 main protease and compared to the recent recognized drug Paxlovid. The screening of 27 algal molecules including 15 oligosaccharides derived from sulfated and non-sulphated polysaccharides, eight pigments and four poly unsaturated fatty acids showed high affinities to interact with the protein active site. Best candidates showing high docking scores in comparison with the reference molecule were sulfated tri-, tetra- and penta-saccharides from Porphyridium sp. exopolysaccharides (SEP). Structural and energetic analyses over 100 ns MD simulation demonstrated high SEP fragments-M pro complex stability. Pharmacokinetics predictions revealed the prospects of the identified molecules as potential drug candidates., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Microalgae are photoautotrophic microorganisms known as producers of a large variety of metabolites. The taxonomic diversity of these microorganisms has been poorly explored. In this study, a newly isolated strain was identified based on the 18S rRNA encoding gene. The phylogenetic analysis showed that the isolated strain was affiliated with the Rhodomonas genus. This genus has greatly attracted scientific attention according to its capacity to produce a large variety of metabolites, including phycoerythrin. Growth and phycoerythrin production conditions were optimized using a Plackett-Burman design and response surface methodology. An expression profile analysis of the cpeB gene, encoding the beta subunit of phycoerythrin, was performed by qRT-PCR under standard and optimized culture conditions. The optimization process showed that maximum cell abundance was achieved under the following conditions: CaCl 2 = 2.1328 g/L, metal solution = 1 mL/L, pH = 7 and light intensity = 145 μmol photons/m 2 /s, whereas maximum phycoerythrin production level occurred when CaCl 2 = 1.8467 g/L, metal solution = 1 mL/L, pH = 7 and light intensity = 157 μmol/m 2 /s. In agreement, positive transcriptional regulation of the cpeB gene was demonstrated using qRT-PCR. This study showed the successful optimization of abiotic conditions for highest growth and phycoerythrin production, making Rhodomonas sp. suitable for several biotechnological applications.

The beneficial properties of algae make them perfect functional ingredients for food products. Algae have a high energy value and are a source of biologically active substances, proteins, fats, carbohydrates, vitamins, and macro- and microelements. They are also rich in polyunsaturated fatty acids, proteins, mycosporine-like amino acids, polysaccharides, polyphenols, carotenoids, sterols, steroids, lectins, halogenated compounds, polyketides, alkaloids, and carrageenans. Different extraction parameters are used depending on the purpose and the substances to be isolated. In this study, the following parameters were used: hydromodule 1:10 and an extraction duration of 1-2 h at the extraction temperature of 25-40 °C. A 30-50% solution of ethanol in water was used as an extractant. Algae extracts can be considered as potential natural sources of biologically active compounds with antimicrobial activity and antiviral properties. The content of crude protein, crude fat, and carbohydrates in U. Prolifera , C. racemosa var. peltata ( Chlorophyta ), S. oligocystum and S. fusiforme (SF-1) was studied. It was found that C. muelleri ( Bacillariophyta ), I. galbana ( Haptophyta ), and T. weissflogii ( Bacillariophyta ) contain about 1.9 times more omega-3 than omega-6 fatty acids. N. gaditana ( Ochrophyta ), D. salina ( Chlorophyta ), P. tricornutum ( Bacillaryophyta ) and I. galbana ( Haptophyta ) extracts showed inhibitory activity of varying intensities against E. coli or P. aeruginosa . In addition, algae and algae-derived compounds have been proposed to offer attractive possibilities in the food industry, especially in the meat sector, to evolve functional foods with myriad functionalities. Algae can increase the biological activity of food products, while the further study of the structure of compounds found in algae can broaden their future application possibilities.

The COVID-19 new variants spread rapidly all over the world, and until now scientists strive to find virus-specific antivirals for its treatment. The main protease of SARS-CoV-2 (M pro ) exhibits high structural and sequence homology to main protease of SARS-CoV (93.23% sequence identity), and their sequence alignment indicated 12 mutated/variant residues. The sequence alignment of SARS-CoV-2 main protease led to identification of only one mutated/variant residue with no significant role in its enzymatic process. Therefore, M pro was considered as a high-profile drug target in anti-SARS-CoV-2 drug discovery. Apigenin analogues to COVID-19 main protease binding were evaluated. The detailed interactions between the analogues of Apigenin and SARS-CoV-2 M pro inhibitors were determined as hydrogen bonds, electronic bonds and hydrophobic interactions. The binding energies obtained from the molecular docking of M pro with Boceprevir, Apigenin, Apigenin 7-glucoside-4'-p-coumarate, Apigenin 7-glucoside-4'-trans-caffeate and Apigenin 7-O-beta-d-glucoside (Cosmosiin) were found to be -6.6, -7.2, -8.8, -8.7 and -8.0 kcal/mol, respectively. Pharmacokinetic parameters and toxicological characteristics obtained by computational techniques and Virtual ADME studies of the Apigenin analogues confirmed that the Apigenin 7-glucoside-4'-p-coumarate is the best candidate for SARS-CoV-2 M pro inhibition.

Sulfated exopolysaccharides (EPS) from Porphyridium cruentum strain were extracted and their antioxidant and anti-bacterial potentials were evaluated based on DPPH free radical, ABTS •+ radical cation and DNA nicking assays, and against four foodborne pathogenic bacteria, respectively. They showed also interesting functional, foaming and emulsion properties. Moreover, microbiological and chemical effects of EPS at 0.5, 1 and 2% on refrigerated minced beef meat were undertaken. Chemical analyses revealed that the treated meat underwent significant decrease (P < 0.05) of primary and secondary lipid oxidation. By the end of the storage period, exopolysaccharides at 2% reduced the metmyoglobin and carbonyl group accumulation compared to control samples and were more efficient (P < 0.05) against microflora proliferation. Furthermore, two multivariate exploratory techniques namely Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA) were applied successfully to all obtained data describing the main characteristics attributed to refrigerated meat samples. Overall, these findings indicated that EPS from P. cruentum are worthy being developed as functional and bioactive components for the meat industry., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Metal elements are widely used in various industrial activities and are considered as common water source contaminants. Thus, the development of cost-effective, simple design and efficient processes for trace metal elements removal from contaminated water sources is of great interest. The effects of cadmium, lead and chromium on growth, biomolecules accumulation and metabolic responses of Amphora coffaeiformis, Navicula salinicola and Dunaliella salina isolated from Tunisian coasts were tested. The bioremediation capacities of the three microalgae strains and the mechanisms involved in ions metal removal were also investigated. N. salinicola and D. salina seem to be better tolerating to Cr, while A. coffaeiformis and N. salinicola showed high resistance to Pb. The expression profile analyses by qRT-PCR of the antioxidant defense-related genes revealed that Cd, Pb and Cr treatments induce the up-regulation of catalase and superoxide dismutase coding genes for A. coffaeiformis and D. salina. Regarding N. salinicola, the catalase coding gene seems to be overexpressed after Cd, Pb and Cr exposure while only Cd and Cr induce superoxide dismutase gene overexpression. Moreover, the phytochelatin synthase (a metal chelator synthesis-related gene) was up-regulated in N. salinicola, A. coffaeiformis and D. salina after Cr exposure and also in A. coffaeiformis and D. salina after Cd exposure. While Pb treatments induce overexpression of phytochelatin synthase coding gene only for D. salina. Studied strains showed promising metal removal efficiencies for both Pb and Cr ions metals reached 95% for D. salina. Ion metal removal mechanisms study revealed that intracellular bioaccumulation process is used by D. salina for Cr up-taking. However, both intracellular and extracellular removal mechanisms are involved for Pb and Cr removal using A. coffaeiformis, N. salinicola and for Pb removal using D. salina. FTIR analysis demonstrated that several functional groups as carboxyl, hydroxyl, amino, phosphate and sulfate may participate in the bioadsorption process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The anti-inflammatory and antihyperglycemic effects of polysaccharides extracted from Alhagi &nbsp; maurorum Medik. seeds, spontaneous shrub collected in Southern of Algerian Sahara were investigated. Their water extraction followed by alcoholic precipitation was conducted to obtain two water-soluble polysaccharides extracts (WSPAM1 and WSPAM2). They were characterized using Fourier transform infrared, 1 H/ 13 C Nuclear Magnetic Resonance, Gas Chromatography-Mass Spectrometry and Size Exclusion Chromatography coupled with Multi-Angle Light Scattering. The capacity of those fractions to inhibit α-amylase activity and thermally induced Bovine Serum Albumin denaturation were also investigated. WSPAM1 and WSPAM2 were galactomannans with a mannose/galactose ratio of 2.2 and 2.4, respectively. The SEC-MALLS analysis revealed that WSPAM1 had a molecular weight of 1.4 × 10 6 Da. The investigations highlighted antinflammatory and antihyperglycemic effects in a dose-dependant manner of WSPAM1 and WSPAM2.

Polysaccharides are complex macromolecules long regarded as energetic storage resources or as components of plant and fungal cell walls. They have also been described as plant mucilages or microbial exopolysaccharides. The development of glycosciences has led to a partial and difficult deciphering of their other biological functions in living organisms. The objectives of glycobiochemistry and glycobiology are currently to correlate some structural features of polysaccharides with some biological responses in the producing organisms or in another one. In this context, the literature focusing on bioactive polysaccharides has increased exponentially during the last two decades, being sometimes very optimistic for some new applications of bioactive polysaccharides, notably in the medical field. Therefore, this review aims to examine bioactive polysaccharide, taking a critical look of the different biological activities reported by authors and the reality of the market. It focuses also on the chemical, biochemical, enzymatic, and physical modifications of these biopolymers to optimize their potential as bioactive agents.

In the last 20 years, xanthophylls from microalgae have gained increased scientific and industrial interests. This review highlights the essential issues that concern this class of high value compounds. Firstly, their chemical diversity as the producer microorganisms was detailed. Then, the use of conventional and innovative extraction techniques was discussed. Upgraded knowledge on the biosynthetic pathway of the main xanthophylls produced by photosynthetic microorganisms was reviewed in depth, providing new insightful ideas, clarifying the function of these active biomolecules. In addition, the recent advances in encapsulation techniques of astaxanthin and fucoxanthin, such as spray and freeze drying, gelation, emulsification and coacervation were updated. Providing information about these topics and their applications and advances could be a help to students and young researchers who are interested in chemical and metabolic engineering, chemistry and natural products communities to approach the complex thematic of xanthophylls.

Karenia selliformis is a marine dinoflagellate responsible for fish-kill events. Its presence has been reported along the Tunisian coasts (south-eastern Mediterranean Sea) since the 1990s. In the present study, a quantitative-PCR assay, based on the internal transcribed spacer (ITS) molecular marker, was developed to detect and quantify K. selliformis in environmental bivalve mollusk samples and in seawater samples. The assay was optimized, and its specificity was confirmed using cross-reactivity experiments against microalgal species commonly found on the Tunisian coasts and/or closely related to K. selliformis. Calibration curves were performed by tenfold dilutions of plasmid DNA harboring target sequence and genomic DNA, attaining a limit of detection of around 5 copies of target DNA per reaction, far below one K. selliformis cell per reaction. The field application of the developed assay showed a powerful detection capability. Thus, the designed assay could contribute to the deployment of in-field diagnostic tools for K. selliformis blooms monitoring., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Amylases are enzymes required for starch degradation and are naturally produced by many microorganisms. These enzymes are used in several fields such as food processing, beverage, and medicine as well as in the formulation of enzymatic detergents proving their significance in modern biotechnology. In this study, a three-stage growth mode was applied to enhance starch production and amylase detection from Chlorella vulgaris. Stress conditions applied in the second stage of cultivation led to an accumulation of proteins (75% DW) and starch (21% DW) and a decrease in biomass. Amylase activities were detected and they showed high production levels especially on day 3 (35 U/ml) and day 5 (22.5 U/ml) of the second and third stages, respectively. The bioinformatic tools used to seek amylase protein sequences from TSA database of C. vulgaris revealed 7 putative genes encoding for 4 α-amylases, 2 β-amylases, and 1 isoamylase. An in silico investigation showed that these proteins are different in their lengths as well as in their cellular localizations and oligomeric states though they share common features like CSRs of GH13 family or active site of GH14 family. In brief, this study allowed for the production and in silico characterization of amylases from C. vulgaris., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Phycoremediation is being considered as an eco-friendly and safe technology for toxics eradication from contaminated aquatic systems. The zinc biosorption capacity of Dunaliella sp. AL-1 was demonstrated. Zinc impacted cell growth and photosynthetic pigments accumulation showing exposure time and concentration-dependent effects. The investigation of the antioxidant protective response to zinc exposition proved a stimulation of guaiacol peroxidase (GPX) activity and an increased rate of total phenolics, flavonoids, condensed tannins and glutathione (GSH). The Box-Behnken design was used to optimize zinc removal conditions by Dunaliella sp. AL-1 strain. The maximum experimental zinc uptake was obtained when zinc concentration, algae dose, initial pH, and contact time were set at 25 mg/L, 0.5 g/L, 7.59 and 13 h 43 min, respectively. Under completely optimized conditions, the fraction of zinc removed intracellularly was much lower than the adsorbed on the cell surface. FTIR analysis Dunaliella sp. AL-1 biomass demonstrated that several functional groups as OH, CH 2 , CO, PO, COO and CO may participate in the biosorption process. A comparative proteomic analysis through nano-HPLC coupled to LC-MS/MS, was performed from pre- and post-zinc treatments cells. Among 199 identified proteins, 60 were differentially expressed of which 41 proteins were down-regulated against 19 up-regulated ones. Target proteins have been demonstrated to be implicated in different metabolic processes mainly photosynthesis and antioxidant defenses., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Polysaccharides from marine algae are one novel source of plant defense elicitors for alternative and eco-friendly plant protection against phytopathogens. The effect of exopolysaccharides (EPS) produced by Porphyridium sordidum on elicitation of Arabidopsis thaliana defense responses against Fusarium oxysporum was evaluated. Firstly, in order to enhance EPS production, a Box-Behnken experimental design was carried out to optimize NaCl, NaNO 3 and MgSO 4 concentrations in the culture medium of microalgae. A maximum EPS production (2.45 g/L) higher than that of the control (0.7 g/L) was observed for 41.62 g/L NaCl, 0.63 g/L NaNO 3 and 7.2 g/L MgSO 4 concentrations. Structurally, the EPS contained mainly galactose, xylose and glucose. Secondly, the elicitor effect of EPS was evaluated by investigating the plant defense-related signaling pathways that include activation of Salicylic or Jasmonic Acid-dependent pathway genes. A solution of 2 mg/mL of EPS has led to the control of fungal growth by the plant. Results showed that EPS foliar application induced phenylalaline ammonia lyase and H 2 O 2 accumulation. Expression profile analysis of the defense-related genes using qRT-PCR revealed the up-regulation of Superoxide dismutases (SOD), Peroxidase (POD), Pathogenesis-related protein 1 (PR-1) and Cytochrome P450 monooxyge-nase (CYP), while Catalase (CAT) and Plant defensin 1.2 (PDF1.2) were not induced. Results suggest that EPS may induce the elicitation of A . thaliana 's defense response against F. oxysporum , activating the Salicylic Acid pathway.

Microalgae have been poorly investigated for new-lipolytic enzymes of biotechnological interest. In silico study combining analysis of sequences homologies and bioinformatic tools allowed the identification and preliminary characterization of 14 putative lipases expressed by Chlorella vulagaris . These proteins have different molecular weights, subcellular localizations, low instability index range and at least 40% of sequence identity with other microalgal lipases. Sequence comparison indicated that the catalytic triad corresponded to residues Ser, Asp and His, with the nucleophilic residue Ser positioned within the consensus GXSXG pentapeptide. 3D models were generated using different approaches and templates and demonstrated that these putative enzymes share a similar core with common α/β hydrolases fold belonging to family 3 lipases and class GX. Six lipases were predicted to have a transmembrane domain and a lysosomal acid lipase was identified. A similar mammalian enzyme plays an important role in breaking down cholesteryl esters and triglycerides and its deficiency causes serious digestive problems in human. More structural insight would provide important information on the enzyme characteristics.

Exopolysaccharide (EPS) from marine microalgae are promising sources of a new generation of drugs. However, lot of them remain to be discovered and tested. In this study, EPS produced by Porphyridium marinum and its oligomers prepared by High Pressure Homogenizer have been tested for different biological activities, i.e., antibacterial, anti-fungal and antibiofilm activities on Candida albicans , as well as for their effects on the viability of murine breast cancer cells. Results have shown that all EPS samples present some biological activity. For antibacterial and antibiofilm activities, the native EPS exhibited a better efficiency with Minimum Inhibitory Concentration (MIC) from 62.5 µg/mL to 1000 µg/mL depending on the bacterial strain. For Candida albicans , the biofilm formation was reduced by about 90% by using only a 31.3 µg/mL concentration. Concerning breast cancer cells, lower molar masses fractions appeared to be more efficient, with a reduction of viability of up to 55%. Finally, analyses of polymers composition and viscosity measurements were conducted on all samples, in order to propose hypotheses involving the activities caused by the intrinsic properties of polymers.

Proteases are industrially important catalysts. They belong to a complex family of enzymes that perform highly focused proteolysis functions. Given their potential use, there has been renewed interest in the discovery of proteases with novel properties and a constant thrust to optimize the enzyme production. In the present study, a novel extracellular neutral protease produced from Arthrospira platensis was detected and characterized. Its proteolytic activity was strongly activated by β-mercaptoethanol, 5,5-dithio-bis-(2-nitrobenzoic acid) and highly inhibited by Hg 2+ and Zn 2+ metal ions which support the fact that the studied protease belongs to the cysteine protease family. Using statistical modelling methodology, the logistic model has been selected to predict A. platensis growth-kinetic values. The optimal culture conditions for neutral protease production were found using Box-Behnken Design. The maximum experimental protease activities (159.79 U/mL) was achieved after 13 days of culture in an optimized Zarrouk medium containing 0.625 g/L NaCl, 0.625 g/L K 2 HPO 4 and set on 9.5 initial pH. The extracellular protease of A. platensis can easily be used in the food industry for its important activity at neutral pH and its low production cost since it is a valuation of the residual culture medium after biomass recovery., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Polysaccharides extracted from marine algae have attracted much attention due to their biotechnological applications, including therapeutics, cosmetics, and mainly in agriculture and horticulture as biostimulants, biofertilizers, and stimulators of the natural defenses of plants. This study aimed to evaluate the ability of alginate isolated from Bifurcaria bifurcata from the Moroccan coast and oligoalginates derivatives to stimulate the natural defenses of tomato seedlings. Elicitation was carried out by the internodal injection of bioelicitor solutions. The elicitor capacities were evaluated by monitoring the activity of phenylalanine ammonia-lyase (PAL) as well as polyphenols content in the leaves located above the elicitation site for 5 days. Alginate and oligoalginates treatments triggered plant defense responses, which showed their capacity to significantly induce the PAL activity and phenolic compounds accumulation in the leaves of tomato seedlings. Elicitation by alginates and oligoalginates showed an intensive induction of PAL activity, increasing from 12 h of treatment and remaining at high levels throughout the period of treatment. The amount of polyphenols in the leaves was increased rapidly and strongly from 12 h of elicitation by both saccharide solutions, representing peaks value after 24 h of application. Oligoalginates exhibited an effective elicitor capacity in polyphenols accumulation compared to alginate polymers. The alginate and oligosaccharides derivatives revealed a similar elicitor capacity in PAL activity whereas the accumulation of phenolic compounds showed a differential effect. Polysaccharides extracted from the brown seaweed Bifurcaria bifurcate and oligosaccharides derivatives induced significantly the phenylpropanoid metabolism in tomato seedlings. These results contribute to the valorization of marine biomass as a potential bioresource for plant protection against phytopathogens in the context of eco-sustainable green technology.

Prorocentrum lima (P. lima) is a widely spread dinoflagellate in the Mediterranean Sea and it has become increasingly involved in harmful algal blooms. The purpose of this study is to develop a probe-based real-time polymerase chain reaction (PCR) targeting the ITS1-5.8S-ITS2 region for the detection and absolute quantification of P. lima based on linear and circular DNA standards. The results have shown that the quantitative PCR (q-PCR), using circular plasmid as a template, gave a threshold cycle number 1.79-5.6 greater than equimolar linear standards. When microalgae, commonly found in aquatic samples were tested, no cross-amplification was observed. The q-PCR brought about a good intra and inter-run reproducibility and a detection limit of 5 copies of linear plasmid per reaction. A quantitative relationship between the cell numbers and their corresponding plasmid copy numbers was attained. Afterwards, the effectiveness of the developed protocol was tested with 130 aquatic samples taken from 19 Tunisian sampling sites. The developed q-PCR had a detection sensitivity of up to 1 cell. All the positive samples were taken from three sampling sites of Medenine Governorate with cell abundances that ranged from 22 to 156,000 cells L -1 of seawater. The q-PCR assay revealed a high sensitivity in monitoring the aquatic samples in which the low concentrations of P. lima were not accurately detected by light microscopy. Indeed, this approach is at the same time rapid, specific and sensitive than the traditional microscopy techniques and it represents a great potential for the monitoring of P. lima blooms., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Quantitative PCR (qPCR) is the method of choice for specific detection and quantification of harmful algal bloom (HAB) species. Development of qPCR assay for simultaneous enumeration of species that frequently co-exist in HABs is required. A high sensitivity TaqMan qPCR assay, using probe and primers, located at ITS1-5.8S-ITS2 rDNA region, detecting, specifically, Karenia selliformis, K. bidigitata, and K. mikimotoi, was designed. ITS1-5.8S-ITS2 rDNA region copy numbers per Karenia cell genome were estimated to 217.697 ± 67.904, allowing cell quantification. An application of the designed methodology in field samples has been conducted, and it showed high sensitivity (detection of around 10 -1 cell/100 mg of bivalve mollusk tissue, equivalent to about 20 copies of the target sequence). We suggest that the optimized method could contribute to early detection of three closely related Karenia species in seafood cultivating areas to promote control quality, guarantee a fast and effective intervention, and improve public health prevention.

Simkania negevensis is an emerging Chlamydia-like bacterium related to human respiratory diseases. An early and accurate detection of this pathogen could be useful to monitor the potential infectious risks and to set suitable outbreak control measures. In Tunisia, distribution and abundance of S. negevensis remain until now largely unknown. In the present work, a qPCR assay, targeting the 16S rRNA gene, for fast detection and quantification of S. negevensis was developed and validated. A high specificity for S. negevensis detection displaying no cross-reaction with the closely related Chlamydia spp. or the other tested microorganisms was noticed. qPCR assay performance was considered very satisfying with detection limits of 5 DNA copies per reaction. qPCR assay validation was performed by screening 37 clinical specimens and 35 water samples. S. negevensis wasn't detected in respiratory samples, but it was found in four cases of water samples. We suggest that the qPCR assay developed in this study could be considered sufficiently characterized to initiate the quantification of S. negevensis in environmental samples., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Bioactive compounds with diverse chemical structures play a significant role in disease prevention and maintenance of physiological functions. Due to the increase in industrial demand for new biosourced molecules, several types of biomasses are being exploited for the identification of bioactive metabolites and techno-functional biomolecules that are suitable for the subsequent uses in cosmetic, food and pharmaceutical fields. Among the various biomasses available, macroalgae are gaining popularity because of their potential nutraceutical and health benefits. Such health effects are delivered by specific diterpenes, pigments (fucoxanthin, phycocyanin, and carotenoids), bioactive peptides and polysaccharides. Abundant and recent studies have identified valuable biological activities of native algae polysaccharides, but also of their derivatives, including oligosaccharides and (bio)chemically modified polysaccharides. However, only a few of them can be industrially developed and open up new markets of active molecules, extracts or ingredients. In this respect, the health and nutraceutical claims associated with marine algal bioactive polysaccharides are summarized and comprehensively discussed in this review.

: The potential reprotoxicity of bifenthrin remains unclear if only the common clinical indicators of reproductive disease are examined. The present study aimed to investigate the efficacy of Spirulina platensis , a microalga rich in antioxidant compounds, against bifenthrin-induced testicular oxidative damage in male mice. At the first, we demonstrate that administration of bifenthrin resulted in a decline of testosterone level and in deterioration of sperm quality that was correlated with significant transcription changes of some specific mRNA and microRNA involved in cholesterol transport, testosterone synthesis, and spermatogenesis. At the biochemical level, we found that oxidative stress was obvious in the bifenthrin group, as evidenced by increase in malondialdehyde (MDA), protein carbonyls (PCO), reactive oxygen species (ROS), and nitrite oxide (NO) that was correlated with activation of genes related to mitochondrial apoptotic signal pathways. We then brought, for the first time to our knowledge, solid and complete experimental evidences that administration of mice with Spirulina extract was sufficient to protect against deleterious effects BF in testicular tissues by abrogating the change in antioxidant enzyme activities; the increase in MDA, PCO, and NO concentrations; and the altered expression level of miRNA and mRNA involved in spermatogenesis. We finally demonstrate that Spirulina restores the production of testosterone in mice as well as epididymal sperm viability and motility. These results suggest a potential antitoxic activity of Tunisian Spirulina deserving further attention., Competing Interests: The authors declare no conflict of interest.