Chemical Characterization and Biological Properties Assessment of Euphorbia resinifera and Euphorbia officinarum Moroccan Propolis.

Although the plants of the genus Euphorbia are largely exploited by therapists in Morocco, the composition and antibacterial activities of propolis from these plants are still unknown. To address this gap, this study aimed to characterize the pollen type, the volatile compounds, and the phenolic and mineral profiles of three Euphorbia propolis samples collected in Morocco and evaluate their antimicrobial activities. The minimum inhibitory concentration of the propolis samples was determined by the microdilution method, and the anti-adherence activity was evaluated by the crystal violet assay. The examination of anti-quorum-sensing proprieties was performed using the biosensor Chromobacterium violaceum CV026. Pollen analysis revealed that Euphorbia resinifera pollen dominated in the P1 sample (58%), while E. officinarum pollen dominated in the P2 and P3 samples (44%). The volatile compounds were primarily composed of monoterpene hydrocarbons, constituting 35% in P1 and 31% in P2, with α-pinene being the major component in both cases, at 16% in P1 and 15% in P2. Calcium (Ca) was the predominant mineral element in both E. resinifera (P1) and E. officinarum (P2 and P3) propolis samples. Higher levels of phenols, flavonoids and dihydroflavonoids were detected in the E. officinarum P2 sample. The minimum inhibitory concentration (MIC) value ranged from 50 to 450 µL/mL against Gram-positive and Gram-negative bacteria. Euphorbia propolis displayed the ability to inhibit quorum sensing in the biosensor C. violaceum CV026 and disrupted bacterial biofilm formation, including that of resistant bacterial pathogens. In summary, the current study evidences the potential use of E. officinarum propolis (P2 and P3) to combat important features of resistant pathogenic bacteria, such as quorum sensing and biofilm formation.

Biochemical characterization of Euphorbia resinifera floral cyathia.

Euphorbia resinifera O. Berg is a plant endemic to the Northern and Central regions of Morocco known since the ancient Roman and Greek times for secreting a poisonous latex containing resiniferatoxin. However, E. resinifera pseudo-inflorescences called cyathia are devoid of laticifers and, therefore, do not secrete latex. Instead, they exudate nectar that local honey bees collect and craft into honey. Honey and cyathium water extracts find a broad range of applications in the traditional medicine of Northern Africa as ointments and water decoctions. Moreover, E. resinifera monofloral honey has received the Protected Geographic Indication certification for its outstanding qualities. Given the relevance of E. resinifera cyathia for bee nutrition, honey production, and the health benefit of cyathium-derived products, this study aimed to screen metabolites synthesized and accumulated in its pseudo-inflorescences. Our analyses revealed that E. resinifera cyathia accumulate primary metabolites in considerable abundance, including hexoses, amino acids and vitamins that honey bees may collect from nectar and craft into honey. Cyathia also synthesize volatile organic compounds of the class of benzenoids and terpenes, which are emitted by flowers pollinated by honey bees and bumblebees. Many specialized metabolites, including carotenoids, flavonoids, and polyamines, were also detected, which, while protecting the reproductive organs against abiotic stresses, also confer antioxidant properties to water decoctions. In conclusion, our analyses revealed that E. resinifera cyathia are a great source of antioxidant molecules and a good food source for the local foraging honeybees, revealing the central role of the flowers from this species in mediating interactions with local pollinators and the conferral of medicinal properties to plant extracts.

Growth stimulation of two legumes (Vicia faba and Pisum sativum) using phosphate-solubilizing bacteria inoculation.

The application of chemical fertilizers for plant growth and protection is one of the reasons for the environment and ecosystem destruction, thus, sustainable agriculture is gaining popularity in research and among farming communities. Although most soils are high in total phosphorus (P), a large portion is unavailable to plants and regarded as a growth-limiting factor. P-solubilizing bacteria (PSB) exploitation is a newly developed bio-solution for enhancing rhizosphere P availability; however, the effect of these bacteria on soil quality and the different phases of plant growth remains unknown. This study aims to evaluate the impact of five strains of PSB, isolated from legume rhizosphere, on the growth of two plants (Vicia faba and Pisum sativum) and certain soil properties. The efficient strains of PSB used are characterized by the P-solubilization, the ACC deaminase activity, the fixation of N, and the IAA, HCN, and siderophores production. The activity of these bacteria is tested in vitro and in vivo under controlled conditions on the growth of the two plants supplemented with the rock P (RP). According to our findings, all PSBs strains outperformed the control in terms of enhancing the growth of the tested legumes with a percentage ranging from 77.78 to 88.88%, respectively. The results showed that all treatments significantly improved plant parameters like nitrogen- (N) and P-content in the plants (67.50, 23.11%), respectively. Also, an increase in the fresh and dry weights of above- (41.17, 38.57%) and below-ground biomasses (56.6, 42.28%), respectively. Compared to the control, this leads to an increase of 72% in root length, 40.91% in plant dry weight, and 40.07% in fresh weight. Rhizospheric soil in PSBs treatments displayed high levels of N, P, and organic matter. All treatments were found to have significantly higher levels of alkaline phosphatase, basal soil respiration, and ß-glucosidase activity than the control. It is concluded that multi-traits PSB can be an alternative for utilizing chemical fertilizers to enhance soil quality and plant growth. Despite the potency of PSBs, its use as a source for the development of sustainable agriculture implies focusing on crop species and adaptation, stress tolerance and climate resilience.

Production, purification, and characterization of inulinase from Streptomyces anulatus.

Inulinase is an enzyme that catalyzes inulin to d-fructose. This enzyme can be extracted from plants, but it is difficult to obtain it in large quantities, so its production cost is high. Therefore, microbial inulinase has great potential for industrial needs. In the last decade, there have been very few reports on actinobacterial inulinases, especially on purification and characterization of inulinase process extraction. This study aims to select actinomycetes that possess high inulinase activity from the soil. To screen inulinase-producing bacteria, modified Czapex-Dox agar supplemented with 1% inulin powder was used. The most effective isolate was Streptomyces sp. EFBO8, morphological and genotypic identification methods, confirmed that the strain is Streptomyces anulatus and that its nucleotide sequence has been deposited in GenBank under accession number OQ073700. To optimize inulinase production, kinetics were performed by using S. anulatus strain, which proved to be most productive with a value of 24,024 EU/mL. The enzyme was purified from the culture filtrate by precipitation with ammonium sulfate (NH4 )2 SO4 , followed by column chromatography Sephadex (G-50) separation. Purified protein has a molecular mass of 3331.83 Da.

Physicochemical Properties and Functional Characteristics of Ecologically Extracted Shrimp Chitosans with Different Organic Acids during Demineralization Step.

The current study aims to develop eco-friendly and economical chitosans with a wide range of applications using organic acids for shrimp shells demineralization. Chitosan samples were extracted from shrimp (Parapenaeus longirostris) shells and the demineralization step was performed with three organic acids (citric, acetic, and lactic) and two mineral acids (hydrochloric and sulfuric). The chitosans were characterized by Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and scanning electron microscopy (SEM). The chitosans' physicochemical properties were also determined. The characteristic bands and functional groups of the chitosans were identified by FTIR spectra. The chitosans' crystallinity order was as follows: ChHCl > ChCitric > ChH2SO4 > ChLactic > ChAcetic. The chitosans' morphological characteristics revealed a smooth surface and fibrous structures with pores. Chitosans extracted by organic acids showed the highest extraction yields. ChHCl and ChCitric had higher degrees of deacetylation values; 83.67% and 81.47%, respectively. The solubility was proportional to the degree of deacetylation. Furthermore, ChH2SO4 and ChCitric had lower molecular weight values; 149 kDa and 183 kDa, respectively. Organic acids are as effective as mineral acids for shrimp shells demineralization. The developed process opens up possibilities to produce chitin and chitosan in a more eco-friendly way and at a lower cost in many industrial sectors.

Isolation and characterization of phosphate solubilizing bacteria naturally colonizing legumes rhizosphere in Morocco.

Low-cost and environmentally friendly agricultural practices have received increasing attention in recent years. Developing microbial inoculants containing phosphate (P) solubilizing bacteria (PSB) represents an emerging biological solution to improve rhizosphere P availability. The present study aims to explore PSB strains isolated from soils located at different bioclimatic stages in Morocco and present in various legumes rhizosphere to improve agronomic microbial fertilizer's effectiveness. It was also aimed to test the isolated strains for their ability to solubilize P in NBRIP medium with Tricalcium P (Ca3 (PO4)2) (TCP), rock phosphate (RP), and their combination as a source of phosphorus, by (22) experiment design. Bacterial strains with a high P solubility index (PSI) were selected, characterized, and compared to commercial control. The vanadate-molybdate method was used to estimate P solubilization activity. Stress tolerance to salinity, acidity, drought, and temperature was tested. From all isolated strains (64), 12 were screened as promising biotechnological interest because of their P solubilization and their good resistance to different drastic conditions. Besides, the strain WJEF15 showed the most P solubility efficiency in NBRIP solid medium with a PSI of 4.1; while the WJEF61 strain was located as the most efficient strain in NBRIP-TCP liquid medium by releasing 147.62 mg.l-1 of soluble P. In contrast, in the NBRIP-RP medium, the strain WJEF15 presented maximum solubilization with 25.16 mg.l-1. The experiment design showed that a combination of RP and TCP with max level progressively increases P solubilization by 20.58%, while the WJEF63 strain has the most efficient concentration of 102.69 mg.l-1. Indeed, among the selected strains, four strains were able to limit tested fungi growth. Thus, results reveal a potential effect of selecting PSBs to support cropping cultures as plant growth-promoting rhizobacteria (PGPR).

Botryosphaeria Dieback (Lasiodiplodia viticola): An Imminent Emerging Threat to the Moroccan Vineyards.

A decline of various grapevines (Vitis vinifera L.) in the province of Doukkala in Morocco was observed in 2021. The causal pathogen was identified as Lasiodiplodia viticola based on morphological characteristics and phylogenetic analysis of the internal transcribed region (ITS), the ß-tubulin gene (TUB) and calmodulin (cmdA). Koch's postulates were confirmed by successful re-isolation of L. viticola from plants inoculated with the pathogen under controlled conditions. The disease was shown to be prevalent in Bni Hilal (71.43%), Laamria (60%), and Boulaouane (40%) districts, but was quasi-absent in Lmechrek. To understand the dominance of L. viticola as one of the grapevine trunk pathogens, effects of temperature (10-40 °C) and pH (pH 3-pH 12) on growth and sporulation were investigated. The species were able to grow in a range of temperatures ranging from 15 to 40°C and showed a higher growth rate at 35 °C. The fungus were also characterized by a broad optimum pH ranging between 3-12. This study is the first report dealing with L. viticola associated with grapevine trunk diseases in Morocco. Additional studies are therefore required to understand the high occurrence of this disease in vineyards, which is likely due to climate changes. A good understanding of this complex disease might help to develop a reliable and sustainable preventive control strategy.

A Panoramic View on Grapevine Trunk Diseases Threats: Case of Eutypa Dieback, Botryosphaeria Dieback, and Esca Disease.

Grapevine trunk diseases (GTD) are currently one of the most devastating and challenging diseases in viticulture, leading to considerable yield losses and a remarkable decline in grapevine quality. The identification of the causal agents is the cornerstone of an efficient approach to fighting against fungal diseases in a sustainable, non-chemical manner. This review attempts to describe and expose the symptoms of each pathology related to GTD, the modes of transmission, and the harmfulness of recently reported agents. Special attention was given to new diagnostic tests and technologies, grapevine defense mechanisms, molecular mechanisms of endophytes fungal colonization, and management strategies used to control these threats. The present extended review is, therefore, an updated state-of-the-art report on the progress in the management of vineyards.

Molecular and clinical assessment of maturity-onset diabetes of the young revealed low mutational rate in Moroccan families.

Background: Maturity-onset diabetes of the young (MODY) is a monogenic form of diabetes characterized by autosomal dominant inheritance. To offer an adequate patient management and therapeutic treatment for MODY patients, in addition to an early efficient diagnosis of their asymptomatic relatives, it is crucial to set an accurate molecular diagnosis. Hence, our aim was to determine the frequency of HNF1A and GCK genes among Moroccan-suspected MODY patients. Methods: Twenty suspected MODY patients were screened for HNF1A and GCK mutations using Sanger sequencing and MLPA methods. Segregation analysis of identified mutations was performed among family members. The pathogenic nature of missense variants was predicted using bioinformatic tools. Results: A total of two mutations were revealed among all patients raising the diagnostic rate to 10%. We identified a large novel GCK deletion (c.209-?_1398+?del) by MLPA in one patient and a previously reported missense substitution (c.92G > A) in HNF1A gene. Conclusion: This is the first investigation to perform the molecular diagnosis of MODY suspected patients. Our findings constitute a primary contribution towards unraveling the genetic landscape involved in the pathogenesis of MODY disease in Morocco.

Comparative Study of the Antioxidant and Enzyme Inhibitory Activities of Two Types of Moroccan Euphorbia Entire Honey and Their Phenolic Extracts.

Honey is a natural food product very famous for its health benefits for being an important source of antioxidant and phenolic compounds. Euphorbia honeys obtained from different regions of Morocco were evaluated for their ability to inhibit acetylcholinesterase, lipoxygenase, tyrosinase and xanthine oxidase activities. Their antioxidant properties were evaluated using the: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging capacity, nitric oxide scavenging activity (NO) and scavenging ability of superoxide anion radical. Then, the phenolic extracts of the same entire honey samples were evaluated by liquid chromatography coupled to diode array detection and mass spectrometry (LC-DAD-MS) and tested for the biological activities previously evaluated on the entire honeys, in order to conduct a comparative study between both (honey and phenolic extracts). The chromatographic profiles for the studied Euphorbia honey extracts were different. Phenolic compounds gallic acid, 4-hydroxybenzoic acid and p-coumaric acid were detected in all samples, whereas kampferol was only present in two samples. Physicochemical parameters and total phenolic content were also determined. Entire honey that recorded the highest rate of phenols was sample M6 (E. resinifera) = 69.25 mg GAE/100 g. On the other hand, the phenolic extracts had better antioxidant and enzyme inhibitory activities than the entire honeys, regardless the monofloral honey type. In conclusion, the studied Euphorbia honeys may have a great potential as antioxidant, anti-inflammatory and anti-tyrosinase sources for pharmaceutical and cosmetic applications.

Antioxidant activity and enzyme inhibitory potential of Euphorbia resinifera and E. officinarum honeys from Morocco and plant aqueous extracts.

Natural products may be applied in a wide range of domains, from agriculture to food and pharmaceutical industries. In this study, the antioxidant properties and the capacity to inhibit some enzymatic activities of Euphorbia resinifera and Euphorbia officinarum aqueous extracts and honeys were assessed. The physicochemical characteristics were also evaluated. Higher amounts of iron, copper and aluminium were detected in E. officinarum honey, which may indicate environmental pollution around the beehives or inadequate storage of honey samples. This honey sample showed higher amounts of total phenols and better capacity for scavenging superoxide anion free radicals and DPPH free radicals as compared with E. resinifera honey, but poorer capacity for inhibiting lipoxygenase, acetylcholinesterase, tyrosinase and xanthine oxidase. The ratio plant mass:solvent volume (1:100) and extraction time (1 - 2 h) were associated with higher total phenols and better antioxidant activities and lipoxygenase, acetylcholinesterase and tyrosinase inhibitory activities, regardless of the plant species. The aqueous extracts had systematically higher in vitro activities than the respective honey samples.

Study of Nutritional and Organoleptic Quality of Formulated Juices from Jujube (Ziziphus lotus L.) and Dates (Phoenix dactylifera L.) Fruits.

The present work aims to elaborate many juice formulas (F1 to F8) from two dried fruits (jujubes: Ziziphus lotus L. and dates: Phoenix dactylifera L.). Physicochemical and biochemical characterization of the formula juices shows that juices rich in dates fruits (F1, F3, F5, and F7) are loaded, on average, in total sugars (129.5 g/l), proteins (3.02 g/l), lipids (1.08 g/l), and carotenoid (0.02 mg/l), while juices rich in jujube fruits (F2, F4, F6, and F8) are overloaded, on average, in phenolic compounds (697 mg/l), flavonoids (6.32 mg/l), condensed tannins (2.1 mg/l), hydrolysable tannins (359.5 mg/l), and viscosity (1.062 mm 10-3 s). All formulations developed have a pH that rotates between 5.12 and 5.20. Total antioxidant capacity (CAT) reveals that formulations F1, F3, F5, and F7 show a strong activity compared to the other formulas. The DPPH test shows that all formulated juices have the same antioxidant profile with IC50 values lower than the template (BHT and Vit C). The FRAP test reveals that F2, F4, F6, and F8 formulas have a strong reducing power. Organoleptic evaluation by a jury shows that F4 formula is the best in terms of odor, aroma, and aftertaste agreeability.

Morphological, Biochemical, and Climatological Analysis of Three Moroccan Henna Verities.

In this study, we aimed to evaluate planted Henna in three sites in Morocco, namely, Alnif, Tafraoute Sidi Ali, and Tazzarine. Morphometric study shows that Tafraoute Sidi Ali Henna variety has highest geometric and weight parameters (length of 27.48 mm, width of 10.92 mm, specific mass of 25.1 mg/leaf, leaf area of 51, 53 mm2, and rib's number of 9.41) when compared to Alnif and Tazzarine varieties. On the other hand, biochemical analysis shows that Tazzarine Henna variety, in the first rank, is characterized by high levels of total sugars (11.27 g/100 g), reducing sugars (5.59 g/100 g), proteins (4.4 g/100g), lipids (3.05 g/100g), phenolic compounds (31.9 g/100 g), flavonoids (5.68 g/100 g), and tannins (5.5 g/100 g). Chromatographic study shows that Tazzarine Henna variety is rich in monocyclic and polycyclic phenolic compounds. Climatic conditions analysis shows that the morphometric and biochemical diversity is related to hydrous and thermal profiles of studied sites. As a conclusion of this work, we can recommend the use of morphometric analysis and phytochemical and chromatographic analysis to determine the quality of Henna in Morocco and elsewhere.

Study of the Biochemical Biodiversity of Camel Milk.

Due to its balanced composition in basic nutrients, its richness in vitamins and in minerals, camel milk has a special place in the daily diet of southern populations. The present study aimed at examining some physicochemical and biochemical characteristics of camel milk (Camelus dromedarius) in Morocco. To achieve this aim, three batches of samples of camel milk were collected from various regions (Dakhla, Errachida, and Fès-Meknes) undergoing physicochemical and biochemical analyses. Results showed that Moroccan camel milk is characterized by slight hydronium potential (pH=6.5), low Dornic acidity (15°D), low density (1.029 kg/l), and high content of ashes (8.06 g/l). Likewise, samples had a high content of macronutrients (Fats: 34.09 g/l; Proteins: 32.4 g/l; Sugar: 49.8 g/l) and micronutrients (Vitamin C: 27.53 mg/l; Flavonoids: 29.05 mg EQ/l: total phenolic compounds: 35.45mg GAE/l). In this respect, working on multiple specimens from different Moroccan regions highlighted an analytical diversity from the south to the north. Comparative study of samples from numerous territories all over the world has confirmed this diversity. North African milk is characterized by high content of proteins, fats, and sugar. On the other hand, oriental milk is peculiarized by high vitamin C content.