Exploring the dynamics of algae-associated microbiome during the scale-up process of Tetraselmis sp. microalgae: A metagenomics approach.

Microalgae have become a key source of valuable compounds, promoting commercial scale applications. However, biological contamination is one of the most critical problems associated with large scale algal production, especially in open systems such as raceway ponds. The current research is the first to assess the effectiveness of open raceway ponds in maintaining a pure culture of Tetraselmis sp., starting from 20 L culture up to 10,000 L culture. Microbial profiling of each successive stage revealed lower abundance of eukaryotic organisms, whereas bacterial abundance increased notably resulting in a significant decrease in Tetraselmis sp. abundance. Furthermore, several bacteria with algae growth-promoting properties were found throughout the various culture stages including Balneola, Roseovarius, and Marinobacter. However, some algae-suppressive bacteria were evidenced at later stages such as Ulvibacter, Aestuariicoccus, and Defluviimonas. Overall, due to the increasing bacterial concentration, considerations limiting bacterial contamination need to be taken.

Synthesis of silver nanoparticles using Bacillus velezensis M3-7 lipopeptides: Enhanced antifungal activity and potential use as a biocontrol agent against Fusarium crown rot disease of wheat seedlings.

Bacillus velezensis M3-7 is a hyperactive mutant, 12-fold improved in its antifungal activity, obtained during a previous study from the wild strain BLB371 after a combination of random mutagenesis and medium component optimization. This study explores the use of this mutant in synthesizing silver nanoparticles (Ag-NPs) for the control of Fusarium crown rot disease (FCR) in wheat seedlings. LC-MS/MS analysis proved that both strains co-produced different families of lipopeptides and that mutagenesis caused the hyper-production of iturin A C14 and C15, the liberation of iturin A C10 and C12, and the inhibition of fengycin release. Our aim was a further improvement in the antifungal activity of the wild strain and the mutant M3-7 in order to control Fusarium crown rot disease (FCR) in wheat seedlings. Therefore, a nanotechnology approach was adopted, and different lipopeptide concentrations produced by the wild strain and the mutant M3-7 were used as capping agents to synthesize silver nanoparticles (Ag-NPs) with enhanced antifungal activity. Ag-NPs formed using 3 mg·mL-1 of the mutant lipopeptides were found to exhibit a good distribution, improved antifungal activity, a promising potential to be used as a biofortified agent for seed germination, and an effective compound to control FCR in wheat seedlings.

Halobacteria-Based Biofertilizers: A Promising Alternative for Enhancing Soil Fertility and Crop Productivity under Biotic and Abiotic Stresses-A Review.

Abiotic and biotic stresses such as salt stress and fungal infections significantly affect plant growth and productivity, leading to reduced crop yield. Traditional methods of managing stress factors, such as developing resistant varieties, chemical fertilizers, and pesticides, have shown limited success in the presence of combined biotic and abiotic stress factors. Halotolerant bacteria found in saline environments have potential as plant promoters under stressful conditions. These microorganisms produce bioactive molecules and plant growth regulators, making them a promising agent for enhancing soil fertility, improving plant resistance to adversities, and increasing crop production. This review highlights the capability of plant-growth-promoting halobacteria (PGPH) to stimulate plant growth in non-saline conditions, strengthen plant tolerance and resistance to biotic and abiotic stressors, and sustain soil fertility. The major attempted points are: (i) the various abiotic and biotic challenges that limit agriculture sustainability and food safety, (ii) the mechanisms employed by PGPH to promote plant tolerance and resistance to both biotic and abiotic stressors, (iii) the important role played by PGPH in the recovery and remediation of agricultural affected soils, and (iv) the concerns and limitations of using PGHB as an innovative approach to boost crop production and food security.

Evidence of the drying technique's impact on the biomass quality of Tetraselmis subcordiformis (Chlorophyceae).

Rapid drying, cost-effective and safe, will increase the viability of using microalgae for several bio-industrial applications. In this study, five different drying techniques of microalgal biomass were investigated. These include freeze drying, oven drying, air drying, sun drying, and microwave drying. Morphology, metabolite content, FAME profiling, chlorophyll content, total organic carbon, and total nitrogen were analyzed. Results showed that the freeze-drying technique preserves the highest amounts of chlorophyll, proteins, and lipids. Oven drying underperformed as it retained the lowest amount of chlorophyll, protein, and lipid content. More importantly, FAME profiling results showed that air drying was the best technique in maintaining the highest amount of polyunsaturated fatty acids and more specifically docosahexaenoic acid (DHA). Furthermore, this process requires the least capital and energy needs. The findings from this study confirmed that the drying technique affects the microalga biomass quality.

Asterarcys quadricellulare (Chlorophyceae) protects H9c2 cardiomyoblasts from H2O2-induced oxidative stress.

Oxidative stress has recently been identified as an important mediator of cardiovascular diseases. The need to find efficient antioxidant molecules is essential in the disease's prevention. Therefore, the present study aimed to evaluate the potential of microalgae bioactive in protecting H9c2 cardiomyoblasts from H2O2-induced oxidative stress. Four microalgal species were investigated for their antioxidant capacity. A qualitative assessment of oxidative stress in H9c2 cardiomyoblasts stained with DCFH-DA, treated with the highly active microalgae extracts, was performed. The protein expression of total caspase-3 was also examined to investigate whether the extract protects H9c2 cardimyoblasts from H2O2-induced apoptosis. High antioxidant activity was observed for the hexanoic extracts after 10 days of cultivation. Asterarcys quadricellulare exhibited the highest antioxidant capacity of 110.59 ± 1.75 mg TE g-1 dry weight and was tested against H9c2 cardiomyoblasts, which were initially subjected to H2O2-induced oxidative stress. This hexanoic extract protected against H2O2 induced oxidative stress with a similar scavenging capacity as N-Acetylcysteine. Furthermore, total caspase-3 was increased following treatment with the hexanoic extract, suggesting that A. quadricellulare also had anti-apoptotic properties. The outcome of our study highlighted the possible use of the local A. quadricellulare strain QUCCCM10 as a natural, safe, and efficient antioxidant to prevent cardiovascular diseases.

Evaluation of Roholtiella sp. Extract on Bell Pepper (Capsicum annuum L.) Yield and Quality in a Hydroponic Greenhouse System.

This study was carried out to investigate the impacts of cyanobacteria (Roholtiella sp.) high-value product extract (HVPE) and water resuspended biomass WRB treatments on bell pepper production using the hydroponic system under greenhouse conditions. Six cyanobacteria treatments (6 ml L-1, 4 ml L-1, and 2 ml L-1 - HVPE, 6 ml L-1, 4 ml L-1, and 2 ml L-1 - WRB, and TR0 as control) were evaluated using the foliar application method. The results showed that foliar application of HVPE with treatments of 2 ml L-1, 4 ml L-1, and 6 ml L-1 produced significantly higher values of physical growth parameters of bell pepper (BP) plants (shoot length, the number of leaves, plant leaf length, plant leaf width, and the diameter of the shoot), SPAD index, yield components (the fruit length, fruit width, the number of fruit per plant, and fresh weight per fruit), biochemical composition [ascorbic acid, phenolic acid, and total soluble solids (TSS)], and the total yield compared to the control group TR0. Also, significant higher values of growth parameters (shoot length, the number of leaves, plant leaf length, plant leaf width, the diameter of the shoot), SPAD index, yield components (the fruit length, fruit width, the number of fruits per plant, and fresh weight per fruit), biochemical composition [ascorbic acid, phenolic acid, and total soluble solids (TSS)], and the total yield were obtained with foliar spraying WRB at 2 ml L-1, 4 ml L-1, and 6 ml L-1 compared to the control group TR0. Consequently, the treated bell pepper with Roholtiella sp. HVPE and WRB were more efficient in enhancing production and chemical constituents compared with the control group.

Marine microbial bioprospecting: Exploitation of marine biodiversity towards biotechnological applications-a review.

The increase in the human population causes an increase in the demand for nutritional supplies and energy resources. Thus, the novel, natural, and renewable resources became of great interest. Here comes the optimistic role of bioprospecting as a promising tool to isolate novel and interesting molecules and microorganisms from the marine environment as alternatives to the existing resources. Bioprospecting of marine metabolites and microorganisms with high biotechnological potentials has gained wide interest due to the variability and richness of the marine environment. Indeed, the existence of extreme conditions that increases the adaptability of marine organisms, especially planktons, allow the presence of interesting biological species that are able to produce novel compounds with multiple health benefits and high economical value. This review aims to provide a comprehensive overview of marine microbial bioprospecting as a growing field of interest. It emphasizes functional bioprospecting that facilitates the discovery of interesting metabolites. Marine bioprospecting was also discussed from a legal aspect for the first time, focusing on the shortcomings of international law. We also summarized the challenges facing bioprospecting in the marine environment including economic feasibility issues.

Genetic engineering to enhance microalgal-based produced water treatment with emphasis on CRISPR/Cas9: A review.

In recent years, the increased demand for and regional variability of available water resources, along with sustainable water supply planning, have driven interest in the reuse of produced water. Reusing produced water can provide important economic, social, and environmental benefits, particularly in water-scarce regions. Therefore, efficient wastewater treatment is a crucial step prior to reuse to meet the requirements for use within the oil and gas industry or by external users. Bioremediation using microalgae has received increased interest as a method for produced water treatment for removing not only major contaminants such as nitrogen and phosphorus, but also heavy metals and hydrocarbons. Some research publications reported nearly 100% removal of total hydrocarbons, total nitrogen, ammonium nitrogen, and iron when using microalgae to treat produced water. Enhancing microalgal removal efficiency as well as growth rate, in the presence of such relevant contaminants is of great interest to many industries to further optimize the process. One novel approach to further enhancing algal capabilities and phytoremediation of wastewater is genetic modification. A comprehensive description of using genetically engineered microalgae for wastewater bioremediation is discussed in this review. This article also reviews random and targeted mutations as a method to alter microalgal traits to produce strains capable of tolerating various stressors related to wastewater. Other methods of genetic engineering are discussed, with sympathy for CRISPR/Cas9 technology. This is accompanied by the opportunities, as well as the challenges of using genetically engineered microalgae for this purpose.

Microalgal-based feed: promising alternative feedstocks for livestock and poultry production.

There is an immediate need to identify alternative sources of high-nutrient feedstocks for domestic livestock production and poultry, not only to support growing food demands but also to produce microalgae-source functional foods with multiple health benefits. Various species of microalgae and cyanobacteria are used to supplement existing feedstocks. In this review, microalgae have been defined as a potential feedstock for domestic animals due to their abundance of proteins, carbohydrates, lipids, minerals, vitamins, and other high-value products. Additionally, the positive physiological effects on products of animals fed with microalgal biomass have been compiled and recommendations are listed to enhance the assimilation of biomolecules in ruminant and nonruminant animals, which possess differing digestive systems. Furthermore, the role of microalgae as prebiotics is also discussed. With regards to large scale cultivation of microalgae for use as feed, many economic trade-offs must be considered such as the selection of strains with desired nutritional properties, cultivation systems, and steps for downstream processing. These factors are highlighted with further investigations needed to reduce the overall costs of cultivation. Finally, this review outlines the pros and cons of utilizing microalgae as a supplementary feedstock for poultry and cattle, existing cultivation strategies, and the economics of large-scale microalgal production.

Marine health of the Arabian Gulf: Drivers of pollution and assessment approaches focusing on desalination activities.

The Arabian Gulf is one of the most adversely affected marine environments worldwide, which results from combined pollution drivers including climate change, oil and gas activities, and coastal anthropogenic disturbances. Desalination activities are one of the major marine pollution drivers regionally and internationally. Arabian Gulf countries represent a hotspot of desalination activities as they are responsible for nearly 50% of the global desalination capacity. Building desalination plants, up-taking seawater, and discharging untreated brine back into the sea adversely affects the biodiversity of the marine ecosystems. The present review attempted to reveal the potential negative effects of desalination plants on the Gulf's marine environments. We emphasised different conventional and innovative assessment tools used to assess the health of marine environments and evaluate the damage exerted by desalination activity in the Gulf. Finally, we suggested effective management approaches to tackle the issue including the significance of national regulations and regional cooperation.

Application of Cyanobacteria (Roholtiella sp.) Liquid Extract for the Alleviation of Salt Stress in Bell Pepper (Capsicum annuum L.) Plants Grown in a Soilless System.

Salinity is one of the abiotic stresses that affect crop growth and productivity in arid and semi-arid regions. Unfortunately, there are few known methods to mitigate the deleterious impacts of salt stress on the development and yield of vegetable crops. Blue-green algae (cyanobacteria) are endowed with the potential to curb the negative impacts of salt stress as they are characterized by biostimulant properties. The present work aimed to investigate the effects of Roholtiella sp. as a foliar extract on the growth characteristics, physiological and biochemical responses of bell pepper (Capsicum annuum L.) plants under varying levels of salinity conditions. A soilless water experiment was carried out in a greenhouse where bell pepper seedlings were grown under five salt concentrations (0, 50, 200, 150, and 200 mM of NaCl). Growth characteristics, pigments content, relative water content, and antioxidant activity (CAT) were determined. Our results showed that growth parameters, relative water content (RWC), chlorophyll a & b concentrations under salinity conditions were negatively affected at the highest concentration (200 mM). Interestingly, the application of Roholtiella sp. foliar extract enhanced the plant growth characteristics as shoot length increased by 17.014%, fresh weight by 39.15%, dry and weight by 31.02%, at various salt treatments. Moreover, chlorophyll a and b increased significantly compared with seedlings sprayed with water. Similarly, RWC exhibited a significant increase (92.05%) compared with plants sprayed with water. In addition, antioxidants activities and accumulation of proline were improved in Roholtella sp. extract foliar sprayed seedlings compared to the plants foliar sprayed with water. Conclusively, at the expiration of our study, the Rohotiella sp. extract-treated plants were found to be more efficient in mitigating the deleterious effects caused by the salinity conditions which is an indication of an enhancement potential of tolerating salt-stressed plants when compared to the control group.

Sustainable Food Production and Nutraceutical Applications from Qatar Desert Chlorella sp. (Chlorophyceae).

Microalgae isolated from the Qatari desert was identified as thermotolerant, with a rich metabolite profile that is appropriate for use as food and health supplements. In this research, a species of Chlorella, QUCCCM3, from the Qatar University Culture Collection of Cyanobacteria and Microalgae, was investigated for its growth characteristics and metabolite compositions for use as potential feedstock for food production. The strain was cultivated at 30, 35, and 40 °C, covering the annual average low and high temperatures in Qatar. The highest growth rates were recorded for cultures at 30 °C with 0.64 ± 0.04 day-1, followed by a growth rate of 0.54 ± 0.06 day-1 at 40 °C, indicating its thermotolerance ability. The biomass exhibited a high protein content (43 ± 2.3%), with existence of lysine (4.13%) as an essential amino acid, and docosahexaenoic acid, linoleic acid, and alpha-linolenic acid as important omega fatty acids present. On the other hand, Chlorella sp. QUCCCM3 also exhibited a high capacity for scavenging free radicals with an antiproliferative effect against chronic myeloid leukemia K562 cancer cells. The results indicate that Chlorella sp. QUCCCM3 is a promising candidate that can be produced year-round, in the Qatar environment, for commercial applications such as feed and nutraceutical supplements.

Algae-Derived Bioactive Compounds with Anti-Lung Cancer Potential.

Lung cancer is one of the major causes of death worldwide. Natural molecules with anti-lung cancer potential are of a great interest and considered as very promising alternative to substitute or enhance the efficiency of the conventional drugs. Recently, algae as source of high value-added compounds are considered as very promising source of these bioactive molecules. These are secondary metabolites that consist mainly of derivatives of peptides, carbohydrates, and lipids with various structures. Accordingly, various mechanisms by which different algae molecules demonstrate attenuation of tumor angiogenesis were stated and discussed. The mode of action of the algae bioactives is closely related to their nature and chemical structure. Furthermore, this literature review considers the synergistic effect between microalgae bioactives and conventional drugs and discuss the economic feasibility of producing microalgae bioactives at large scale to conclude with some future perspectives related to algae-based drug discovery.

Overproduction of the Bacillus thuringiensis Vip3Aa16 toxin and study of its insecticidal activity against the carob moth Ectomyelois ceratoniae.

The vip3Aa16 gene of Bacillus thuringiensis strain BUPM95 was cloned and expressed in Escherichia coli. Optimization of Vip3A16 protein expression was conducted using Plackett-Burman design and response surface methodology. Accordingly, the optimum Vip3A16 toxin production was 170µg/ml at 18h post-induction time and 39°C post-induction temperature. This corresponds to an improvement of 21times compared to the starting conditions. The insecticidal activity, evaluated against Ectomyelois ceratoniae, displayed an LC50 value of 40ng/cm(2) and the midgut histopathology of Vip3Aa16 fed larvae showed vacuolization of the cytoplasm, brush border membrane destruction, vesicle formation in the apical region and cellular disintegration.

Histopathological effects and determination of the putative receptor of Bacillus thuringiensis Cry1Da toxin in Spodoptera littoralis midgut.

Bacillus thuringiensis subsp. aizawai strain HD133, known by its effectiveness against Spodoptera species, produces many insecticidal proteins including Cry1Ab, Cry1Ca and Cry1Da. In the present study, the insecticidal activity of Cry1Da against Spodoptera littoralis was investigated. It showed toxicity with an LC(50) of 224.4 ng/cm(2) with 95% confidence limits of (178.61-270.19) and an LC(90) of 467.77 ng/cm(2) with 95% confidence limits of (392.89-542.65). The midgut histopathology of Cry1Da fed larvae showed vesicle formation in the apical region, vacuolization and destruction of epithelial cells. Biotinylated-activated Cry1Da toxin bound protein of about 65 kDa on blots of S. littoralis brush border membrane preparations. This putative receptor differs in molecular size from those recognized by Cry1C and Vip3A which are active against this polyphagous insect. This difference in midgut receptors strongly supports the use of Cry1Da as insecticidal agent, particularly in case of Cry and/or Vip-resistance management.

Investigation of antimicrobial activity and statistical optimization of Bacillus subtilis SPB1 biosurfactant production in solid-state fermentation.

During the last years, several applications of biosurfactants with medical purposes have been reported. Biosurfactants are considered relevant molecules for applications in combating many diseases. However, their use is currently extremely limited due to their high cost in relation to that of chemical surfactants. Use of inexpensive substrates can drastically decrease its production cost. Here, twelve solid substrates were screened for the production of Bacillus subtilis SPB1 biosurfactant and the maximum yield was found with millet. A Plackett-Burman design was then used to evaluate the effects of five variables (temperature, moisture, initial pH, inoculum age, and inoculum size). Statistical analyses showed that temperature, inoculum age, and moisture content had significantly positive effect on SPB1 biosurfactant production. Their values were further optimized using a central composite design and a response surface methodology. The optimal conditions of temperature, inoculum age, and moisture content obtained under the conditions of study were 37°C, 14 h, and 88%, respectively. The evaluation of the antimicrobial activity of this compound was carried out against 11 bacteria and 8 fungi. The results demonstrated that this biosurfactant exhibited an important antimicrobial activity against microorganisms with multidrug-resistant profiles. Its activity was very effective against Staphylococcus aureus, Staphylococcus xylosus, Enterococcus faecalis, Klebsiella pneumonia, and so forth.

Characterization of Tunisian Bacillus thuringiensis strains with abundance of kurstaki subspecies harbouring insecticidal activities against the lepidopteran insect Ephestia kuehniella.

The study of 257 crystal-producing Bacillus thuringiensis isolates from bioinsecticide free soil samples collected from different sites in Tunisia, was performed by PCR amplification, using six primer pairs specific for cry1, cry2, cry3, cry4, and vip3A genes, by the investigation of strain plasmid pattern, crystal morphology and delta-endotoxin content and by the assessment of insecticidal activities against the lepidopteran insect Ephestia kuehniella. Based on plasmid pattern study, 11 representative strains of the different classes were subjected to morphological and molecular analyses. The comparison of the PFGE fingerprints confirmed the heterogeneity of these strains. B. thuringiensis kurstaki strains, harbouring at the same time the genes cry1A, cry2, cry1Ia, and vip3A, were the most abundant (65.4%). 33.34% of the new isolates showed particular delta-endotoxin profiles but no PCR products with the used primer sets. B. thuringiensis israelensis was shown to be also very rare among the Tunisian B. thuringiensis isolates diversity. These findings could have considerable impacts for the set up of new pest control biological agents.

Evidence of the involvement of E358, A498 and C571 of a new Cry1Ac delta-endotoxin of Bacillus thuringiensis in its high insecticidal activity against Ephestia kuehniella.

A new cry1Ac-type gene was cloned from Bacillus thuringiensis strain BLB1, sequenced and expressed. The deduced amino acid sequence of the polypeptide has a predicted molecular mass of 132.186 kDa. The amino acid sequence alignment of BLB1 Cry1Ac with those of the published ones showed that this is a new delta-endotoxin. When compared with Cry1Ac of Bacillus thuringiensis strain HD1, it was found that BLB1 Cry1Ac harbours three mutations: V358E localized in domain II and V498A and Y571C localized in domain III. When the BLB1 Cry1Ac toxin was expressed in an acrystalliferous strain of B. thuringiensis (HD1CryB), bipyramidal crystals were produced. The spore-crystal mixture of this recombinant strain was at least two-fold more active against larvae of the lepidopteran Ephestia kuehniella than that of the recombinant strain expressing Cry1Ac of HD1. The study of the structural effect of these mutations suggested that they may stabilize key regions involved in the binding of the domains II and III to insect receptors.

Improvement of Bacillus thuringiensis bacteriocin production through culture conditions optimization.

BUPM4 is a Bacillus thuringiensis subsp. kurstaki strain, isolated from Tunisian soil, producing an original bacteriocin named Bacthuricin F4. The optimization of the latter production conditions was carried out under several physicochemical conditions. It was found that the highest bacteriocin activity was reached at low aeration while bacteriocin synthesis yields were strongly reduced at higher ones. A balance between growth and bacteriocin synthesis, both highly dependent on aeration, was taken into account for the overproduction of bacteriocin. Both glucose and glycerol were shown to be necessary for Bacthuricin F4 maximal synthesis. In addition, the optimal carbon/nitrogen ratio for bacteriocin production is 9. In such optimal conditions, more than 4-fold greater bacteriocin production was obtained than when using TSB medium.

A new Tunisian strain of Bacillus thuringiensis kurstaki having high insecticidal activity and delta-endotoxin yield.

BLB1 is a new Bacillus thuringiensis kurstaki strain, isolated from a Tunisian soil sample. Assay of toxicity of BLB1 crystal proteins resulted in an LC50 of 70.32 ng of toxin per mg of flour against third instar Ephestia kuehniella with confidence limits of (31.6-109.04 ng). This LC50 is less than that of the commercial strains HD1 used as a reference. The characterization of this strain by scanning transmission electron microscopy, analysis of its cry genes content by PCR-sequencing, and analysis of its delta-endotoxin patterns demonstrate that it belongs to the same subgroup than HD1, but ruled out the involvement of cry gene content or protoxin activation in the hypertoxicity of this strain. Taking into account the delta-endotoxin/spore ratio for each strain, and by allowing the estimation of the production level per spore, it might be concluded that BLB1 production is the highest, when compared with that of HD1. On the basis of its toxicity, BLB1 could be considered as a strain of great interest and would allow the production of quantities of bioinsecticides at low cost.