Exploring Technological, Safety and Probiotic Properties of Enterococcus Strains: Impact on Rheological Parameters in Fermented Milk.

Enterococci, known for their resilience, are commonly found in food, the environment, and the gastrointestinal tracts of humans and animals. In recent research, six strains of enterococcus were isolated from bat guano. These include Enterococcus mundtii SRBG1, Enterococcus gallinarum SRBG3, Enterococcus faecium SRBG2, Enterococcus casseliflavus EC1, and Enterococcus devriesei CAU 1344. Identification was done using 16S DNA analysis. Each strain underwent evaluation for its technological properties (such as tolerances to various NaCl concentrations and temperatures, as well as amylolytic, ß-galactosidase, lipolytic, and proteolytic activities, and EPS production) and selected probiotic properties (including safety profile, resistance to 0.3 percent bile salts and gastric juice with a pH of 2.5, lysozyme tolerance, and antibacterial and antibiofilm activities against four foodborne pathogens). The results were analyzed using Principal Component Analysis. This analysis revealed that E. mundtii SRBG1 and E. gallinarum SRBG3, followed by E. faecium SRBG2, were most closely associated with a broad range of technological characteristics and were subsequently used for fermenting skimmed milk. The rheological properties of the samples indicated a shear-thinning or non-Newtonian behavior. Furthermore, during storage of the fermented milk at 4 °C over periods of 1, 7, 14, and 21 days, there were no significant changes in bacterial count (at around 7 log10 CFU/mL) and pH when fermented with the three evaluated strains.

The Impact of Origanum vulgare Supplementation on Human Asthenozoospermic Sperm Parameter Quality.

Male infertility is a complex multifactorial disease and a real health problem; 50% of infertile men have identifiable causes detectable by fundamental sperm analysis. Numerous research studies have shown the possibility of treating abnormal semen samples with some drugs before artificial insemination, yet evidence of the drug's effectiveness remains minimal. In our previous work, we tested the effectiveness of some essential oils, such as eucalyptus (Eucalyptus globulus Labill.), oregano (Origanum vulgare L.), and sage (Salvia officinalis L.) on sperm parameters. The essential oil of oregano showed the best ameliorative effect. In present, we examined the effect of the essential oil of O. vulgare on the physiological parameters and the specific activity of certain antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), lipid peroxidation rate (MDA), and GAPDH and antioxidant and metabolic biomarkers, characterizing the quality of human sperm. The results showed that in vitro supplementation of oregano significantly improves the mobility and antioxidant activities, without harmful effects on the integrity of the sperm's DNA, and that the selected concentration of oregano EO is nontoxic and may be considered a therapeutic alternative to heal sperm motility problems in asthenozoospermic patients.

Essential oils from Dysphania genus: Traditional uses, chemical composition, toxicology, and health benefits.

The genus Dysphania belongs to the Amaranthaceae family and is known for its many health benefits. Therefore, it is commonly available worldwide and includes more than 47 species, five species have been mainly reported, and D. ambrosioides has been one of the most widely used plants for thousands of years as a remedy for a wide range of ailments. In recent investigations, the essential oils of the genus Dysphania have been examined for their antibacterial, antioxidant, and antiviral properties related to specific components such as terpenoid compounds that exhibit pharmacological activity. Moreover, some of Dysphania's compounds show a toxicological effect. Therefore, the objective of the study was to provide EO chemical composition and pharmacological data of the genus Dysphania.

Effect of encapsulated probiotic in Inulin-Maltodextrin-Sodium alginate matrix on the viability of Enterococcus mundtii SRBG1 and the rheological parameters of fermented milk.

In the current research, Enterococcus mundtii SRBG1 newly isolated from Bat guano was encapsulated using spray drying technique to create a probiotic powder using six combinations of inulin, maltodextrin and sodium alginate. The encapsulation yield, moisture content, physical characteristics, and shape were investigated. Microcapsules yields ranged from 67 to 85 percent, which is consistent with typical B-290 spray-drier yields. The moisture content showed to increase (4 ± 0.15%) with the addition of sodium alginate to inulin and maltodextrin. In the gastrointestinal conditions (simulated gastric juice and bile salts), it was shown that the viability of probiotic cells in capsules was higher than that of free cells. This demonstrated the effectiveness of combining inulin and maltodextrin to encapsulate substances in surviving in gastro-intestinal conditions. Additionally, we evaluated the non-encapsulated and encapsulated SRBG1 by assessing their impact on the rheological parameters of fermented milk. The results showed that in the absence of sodium alginate the viscosity of milk was lower than with the other protectors, which was confirmed by the quick acidification of the fermented milk by microcapsules containing sodium alginate.

Optimisation and characterization of α-D-glucan produced by Bacillus velezensis RSDM1 and evaluation of its protective effect on oxidative stress in Tetrahymena thermophila induced by H2O2.

A novel exopolysaccharide was isolated from Bacillus velezensis RSDM1, purified and characterized. The optimization was carried out by Box-Behnken design and the highest yield of EPS-RSDM1 was 1969.53 mg/L. The EPS was composed by glucose with a molecular weight of 1.78 × 104 Da and connected by α-1,6 and α-1,3. The EPS was thermally stable and showed pseudoplastic rheology. The SEM have demonstrated a porous structure. Furthermore, EPS-RSDM1 showed a protective effect against the oxidative stress induced by H2O2 in Tetrahymena thermophila. The exposure of the cells to H2O2 reduced significantly the cell survival and CAT, SOD, GR, SDH and GAPDH activities, and increased the MDA production. However, pretreatment of the cells with EPS-RSDM1 prior to H2O2 exposure elevated the cell survival and enzyme activities, and decreased the level of MDA activity. Our study suggest that EPS-RSDM1 could be useful for preventing cellular oxidation in pharmaceutical and food industries.

Isolation, Molecular Identification and Antibacterial Potential of Marine Bacteria from Deep Atlantic Ocean of Morocco.

Background: Antibiotic resistance is an important concern for the public health authorities at global level. It is detrimental to human and environmental ecosystems, thus, there is a big need for natural bioactive compounds. In this work, we aimed to find out biomolecules derived from marine bacteria that may constitute an alternative to antibiotics. Methods: We isolated and identified thirty one marine bacteria collected from deep ocean water in central coast of Safi city, Morocco. Then, we induced biomolecules production in six marine bacterial strains. The extracts were tested for their antibacterial activity against gram-negative and gram-positive bacteria such as Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 33592 and Listeria monocytogenes ATCC 19117. Furthermore, we partially analyzed the chemical composition of these biomolecules and evaluated their sensibility to different temperatures. Results: The six marine bacteria were able to produce molecules which inhibited the three pathogenic strains with high inhibition zones reaching 27 mm. These molecules were characterized by heat stability from 60 to 121°C relying on each strain. Conclusion: The produced molecules may offer a great potential to pharmaceutical industries as they may constitute an alternative to antibiotics that are becoming less effective due to the emergence of drugs resistance.

Screening, optimization and characterization of exopolysaccharides produced by novel strains isolated from Moroccan raw donkey milk.

Two exopolysaccharides (EPS) producing strains, isolated from raw donkey milk were identified as Leuconostoc mesenteroides SL and Enterococcus viikkiensis N5 using 16S rDNA sequencing. The Box Benheken design exhibited the highest yield of EPS-SL (672.342 mg/L) produced by SL and of EPS-N5 (901 mg/L) produced by N5. The molecular weight was 1.68×104 for EPS-SL and 1.55×104 Da for EPS-N5. FTIR, NMR and GC-MS analysis showed that the EPS are heteropolysaccharides. The SEM image showed that the EPS-SL was smooth and represented a lotus leaf shape and EPS-N5 revealed a stiff-like, porous appearance and was more compact than EPS-SL. The TGA analyses showed high thermal stability and degradation temperature. Additionally, the two EPSs possessed antibacterial and antioxidant activity, and the EPS-SL had the stronger antioxidant activity. Consequently, these results suggest that the functional and biological properties of EPS-SL and EPS-N5 imply the potential application in the food and pharmaceutical industries.

Comparability of CMV DNA Extraction Methods and Validation of Viral Load.

Human cytomegalovirus is a herpesvirus that has a worldwide seroprevalence of more than 60% of adults in developed countries and 90% in developing countries. Severe disabilities in newborns are characteristic of the human cytomegalovirus congenital infection, and this virus is implicated in graft rejection in transplant patients. To treat and follow-up the infection, the CMVPCR viral loads are required, and the DNA extraction step remains very important; however, the quantity, quality, and purity of extracted DNA from different biological fluids influence the results of PCR amplification, that is why for reliable results, the choice of nucleic acid extraction methods requires careful attention. Materials and methods: In this study, we compare 4 protocols, I (EZ1 DSP Virus kit), II (EZ1 Virus mini kit), III (QIAamp DSP virus kit), and IV (heating); the extractions are made from plasma collected on EDTA tubes, and the concentration of extracted DNA was measured on NanoDrop Lite followed by real-time CMVPCR using an Artus CMV QS-RGQ kit. All protocols are performed following the manufacturer's instructions. Results: This study is conducted on the samples of 135 transplant patients whose follow-up medical tests related to human cytomegalovirus infection; since most of the CMVPCR results are negative, we have chosen the 10 CMVPCR positive samples and 2 negative samples as controls to conduct this comparison study. By using NanoDrop Lite to evaluate the DNA concentration, the yield of extracted DNA is higher in our heating protocol than other protocols, the EZ1 DSP virus kit and EZ1 Virus mini kit show homogeneous quantities, and the QIAamp DSP virus kit shows very low DNA yields. Comparing cycle threshold and viral loads by real-time PCR, all these protocols identified negative samples (100%), and the previously positive samples used were as follows: protocol IV (90%), protocol II (60%), and protocol I (40%). QIAamp DSP virus kit results were not real-time PCR applicable and were non-conclusive because of the low DNA yields. Conclusion: Our developed heating method (protocol IV) is very effective, reliable, simple, fast, and cheap compared to the other protocols in our study.

Isolation and Identification of Dominant Bacteria from Raw Donkey Milk Produced in a Region of Morocco by QIIME 2 and Evaluation of Their Antibacterial Activity.

Recently, the interest in donkey milk has increased considerably because it proved high nutritive and functional values of their ingredients. Its chemical composition is widely studied, but its microbiota, especially lactic acid bacteria, remains less studied. This study focuses on analyzing, isolating, and identifying lactic acid bacteria and evaluating their capacity to produce biomolecules with antibacterial activity. Among 44 strains identified, 43 are Gram-positive, and most are catalase-negative and cocci-shaped. Five strains were selected to evaluate their antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and Escherichia coli. Different induction methods allowed to amplify the antibacterial effects against these pathogenic strains.

Insights on Health and Food Applications of Equus asinus (Donkey) Milk Bioactive Proteins and Peptides-An Overview.

Due to its similarity with human milk and its low allergenic properties, donkey milk has long been used as an alternative for infants and patients with cow's milk protein allergy (CMPA). In addition, this milk is attracting growing interest in human nutrition because of presumed health benefits. It has antioxidant, antimicrobial, antitumoral, antiproliferative and antidiabetic activity. In addition, it stimulates the immune system, regulates the gastrointestinal flora, and prevents inflammatory diseases. Although all donkey milk components can contribute to functional and nutritional effects, it is generally accepted that the whey protein fraction plays a significant role. This review aims to highlight the active proteins and peptides of donkey milk in comparison with other types of milk, emphasizing their properties and their roles in different fields of health and food applications.

Effect of Origanum vulgare Essential Oil Supplementation on the Advanced Parameters of Mobility and on the Integrity of Human Sperm DNA.

The reduced sperm mobility is one of the most important causes of male infertility. Several reports have indicated that the treatment of subnormal sperm samples with certain agents prior to artificial insemination significantly improves the fertilizing potential of sperm. We have among others some stimulants such as pentoxifylline, relaxin, prostaglandin E, and diltiazem. In our precedent work, we had tested the effect of supplementation with three essential oils, namely, sage (Salvia officinalis), oregano (Origanum vulgare), and eucalyptus (Eucalyptus globulus), on sperm cell mobility and vitality. Oregano oil had shown interesting biological properties by giving the best values of progressive mobility and vitality. In this study, we aim to verify the effect of oregano oil supplementation on the advanced parameters of mobility and on the integrity of the sperm DNA of 25 male infertile volunteers. Our results showed that oregano oil over an incubation period of 5 to 10 min of exposure significantly improves the advanced parameters of mobility, namely, curvilinear velocity (VCL), linear velocity (VSL), the mean velocity of the path (VAP), and the amplitude of the displacement (ALH). The effect of the increase in the VCL decreased the linearity (LIN), the mean line (STR), and the mean wobble (WOB). Oregano oil at 5 min had no significant effect on the DNA fragmentation index (DFI) and sperm decondensation index (SDI). However, at 10 min, it had a significant effect on both DFI and SDI. The analysis of our results showed that this plant oil rich in terpenoids and phenolic antioxidants could be a quite good in vitro additive with high potential for the world of medically assisted reproduction.

Assessment of the electrochemical behaviour of Nickel-Titanium-based orthodontic wires: Effect of some natural corrosion inhibitors in comparison with fluoride.

BACKGROUND: The aim of this study is to assess the corrosion resistance behaviour of Nickel-Titanium-based orthodontic wires (NiTi) in different concentrations of Sodium Fluoride (NaF) and the corrosion's inhibitory effect of the extracts of some medicinal plants (essential oils, hydrosols and extract). MATERIAL AND METHODS: In this study we used NiTi (3M) and CuNiTi (ORMCO, 35°C, California) orthodontic wires. The following electrolytes were prepared: Lactate Ringer solution with additions of 0.1%, 0.5% or 1% of Sodium Fluoride and the extracts of different plants: Artemisia, Syzygium aromaticum (Clove) and Celtis australis. Corrosion resistance was studied using anodic potentiodynamic polarisation and electrochemical impedance spectroscopy measurements. At the end of the experiment, microscopic images of wires were performed. ANOVA test with the comparison of Bonferroni and Tukey tests were performed to elucidate comparisons among all groups. RESULTS: The higher sodium fluoride concentration is related to negative corrosion potential for both NiTi and CuNiTi orthodontic wire. Hydrosols are associated to positive values of corrosion potential. CuNiTi has a lower corrosion resistance than NiTi. CONCLUSIONS: The prescription of toothpastes containing sodium fluoride should be reduced especially for patients wearing fixed orthodontic appliances. Eugenol may be considered as alternative of sodium fluoride for orthodontic patients for its anti-microbial and anti-corrosive effects. Key words:Corrosion behaviour, Sodium Fluoride, Nickel-Tatanium, orthodontic wires, corrosion inhibitors, aromatical plants.

The Effect of Supplementation with Some Essential Oils on the Mobility and the Vitality of Human Sperm.

The objective of this work is to study the improvement effect of some essential oils of sage (Salvia officinalis), oregano (Origanum vulgare), and eucalyptus (eucalyptus globulus) on the physiological parameters characterizing the quality of human sperm (mobility and vitality). We find natural biomolecules to improve sperm quality to increase the chances of success of very low in vitro fertilization (IVF) that stagnate around 20%. Sperm samples were mixed with different concentrations of essential oils. The effect of these essential oils on the motility and vitality of spermatozoa has been analyzed. The mobility was determined using a Computer Assisted Sperm Analysis (CASA). In the other side, the evaluation of sperm vitality was performed by staining eosin 2% and the microscopic examination is carried out via optical microscope. A drop of sperm will be mixed with a drop of eosin solution 2%, spread between the slip and coverslip, then allowed to air dry, and examined under a microscope. A significant improvement in the mobility and vitality of human spermatozoa has been noted with oregano. Eucalyptus after 10 min of exposure also significantly improves the mobility and vitality of the spermatozoa. Sage does not improve mobility for these incubation times but significantly improves vitality.

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.

Consequences of Androctonus mauretanicus and Buthus occitanus scorpion venoms on electrolyte levels in rabbits.

Androctonus mauretanicus (A. mauretanicus) and Buthus occitanus (B. occitanus) scorpions, which belong to the Buthidae family, are the most venomous scorpions in Morocco. For the first time, we investigated the effects of such scorpion venoms on serum electrolytes in subcutaneously injected rabbits. For this purpose, 3 groups of 6 albinos adult male rabbits (New Zealand) were used in this experiment. Two of the groups were given a single subcutaneous injection of either crude Am venom (5 µg/kg) or Bo venom (8 µg/kg) whereas the third group (control group) only received physiological saline solution (NaCl 0.9%). The blood samples were collected from injected rabbits via the marginal vein at time intervals of 30 min, 1 h, 2 h, 4 h, 6 h and 24 h after venom injection. The concentrations of electrolytes in the serum samples were measured. Our study indicates that scorpion envenomation in vivo, rabbit animal model, caused severe and persistent hypomagnesaemia and hypochloremia, which are accompanied of hypernatremia, hyperkalemia and hypercalcaemia. The intensity of electrolytes imbalance was clearly superior in the case of A. mauretanicus scorpion venom (although a lower quantity of venom was injected). This is coherent with the experimental data which indicate that A. mauretanicus venom is more toxic than B. occitanus venom.

Assessment of GAPDH expression by quantitative real time PCR in blood of Moroccan AD cases.

INTRODUCTION: Neuroproteomics studies have showed the high affinity interactions between GAPDH - ß-amyloid in Alzheimer disease. The aim of our study is to complete our previous studies by assessing the mechanism responsible of decreased expression of GAPDH protein in the blood of Moroccan AD cases probably due to an alteration at the transcriptional level or at the post translational level. METHODS: The mRNA expression of GAPDH was assessed by quantitative real time PCR in AD cases and healthy controls. RESULTS: Our result revealed a significant difference of mRNA expression level of GAPDH in AD cases as compared to healthy controls (P<0.05). CONCLUSION: This data is consistent with several studies by showing the direct involvement of GAPDH in amyloid aggregation by undergoing several modifications, which influence its chemical structure and its biological activity.

Peroxiredoxin-2 up-regulation in inflammatory bowel disease: Friend or foe?

BACKGROUND: Inflammatory bowel diseases (IBD) are chronic multi-factorial inflammatory disorders. Accumulating investigations have provided compelling evidence that describe the interplay of a complex genetic landscape and inappropriate inflammatory response to intestinal microbes in disease etiopathogenesis but still pose challenges in diagnostic practices. METHOD: In this study, comparative proteomic analysis was conducted to identify disease specific proteins underlying IBD pathogenetic mechanisms. Total blood proteins of the IBD patients and healthy subjects were analyzed with one-dimensional electrophoresis; differentially expressed bands were excised and subjected to matrix-assisted laser desorption ionization-time of flight mass spectrometry along with nanoflow liquid chromatography electrospray ionization-tandem mass spectrometry analysis. Presence of glycosylation, hydroxylation, and phosphorylation post-translational modifications was further investigated by immunoprecipitation. RESULTS: Peroxiredoxin-2 (PRDX2) and hemoglobin-subunits proteins, which are closely involved in the response to oxidative stress, were identified. PRDX2 was selected for further validation using western blot and reverse transcription-polymerase chain reaction. PRDX2 overexpression was restricted to the protein level within the membrane fraction. Immunoprecipitation identified PRDX2 to be post-translationally glycosylated and phosphorylated. CONCLUSION: Our findings demonstrate the implication of PRDX2 in IBD. Future studies are required to establish its functional role and to determine the clinical utility.

A proteomic approach for the involvement of the GAPDH in Alzheimer disease in the blood of Moroccan FAD cases.

Several articles have highlighted the potential involvement of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in neurodegeneration by showing a non-glycolytic activity of GAPDH specifically in the brains of subjects with Alzheimer's disease (AD). The novel aim of this study was to elucidate the critical role of GAPDH and its interaction with ß-amyloid in the blood of Moroccan patients with familial AD (FAD) carrying presenilin mutations and in sporadic late onset AD (LOAD). Our results show a significant decrease in the activity of GAPDH in blood samples from patients with FAD as compared to sporadic cases and healthy controls. The expression level of GAPDH in brain specimens from mutant tau transgenic mice and patients with FAD was unchanged as compared to healthy controls. In contrast, the expression level of GAPDH in blood samples from mutant tau transgenic mice and patients with FAD was decreased as compared to sporadic cases and healthy controls. Moreover, there is an accumulation of ß-amyloid aggregates in the blood samples of patients with FAD and an increase in amyloid fibrils in both the blood and brain samples of these patients. Our study adds new insight to previous ones by showing the involvement of GAPDH in AD, which may influence the pathogenesis of this neurodegenerative disease.

Preliminary Study to Assess the Performance of Mengovirus Elution from Sludge.

In the virus detection protocol for sludge, the viral elution step from solids to solution is critical. In this study, mengoviruses were detected in artificially contaminated sludge with a qRT-PCR assay. The viral yields ranged between 19 and 66 % for 60 % sludge. This study demonstrates that mengovirus can be used as a sample process control for analysis of sewage sludge.

Cloning and heterologous overexpression of three gap genes encoding different glyceraldehyde-3-phosphate dehydrogenases from the plant pathogenic bacterium Pseudomonas syringae pv. tomato strain DC3000.

The gammaproteobacterium Pseudomonas syringae pv. tomato DC3000 is the causal agent of bacterial speck, a common disease of tomato. The mode of infection of this pathogen is not well understood, but according to molecular biological, genomic and proteomic data it produces a number of proteins that may promote infection and draw nutrients from the plant. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a major enzyme of carbon metabolism that was reported to be a surface antigen and virulence factor in other pathogenic microorganisms, but its possible role in the infection process of P. syringae has so far not been studied. Whole-genome sequence analyses revealed the occurrence in this phytopathogenic bacterium of three paralogous gap genes encoding distinct GAPDHs, namely two class I enzymes having different molecular mass subunits and one class III bifunctional D-erythrose-4-phosphate dehydrogenase/GAPDH enzyme. By using genome bioinformatics data, as well as alignments of both DNA and deduced protein sequences, the three gap genes of P. syringae were one-step cloned with a His-Tag in pET21a vector using a PCR-based strategy, and its expression optimized in Escherichia coli BL21 to achieve high yield of the heterologous proteins. In accordance with their distinct molecular phylogenies, these bacterial gap genes encode functional GAPDHs of diverse molecular masses and nicotinamide-coenzyme specificities, suggesting specific metabolic and/or cellular roles.