Mycotoxins occurrence in milk and cereals in North African countries - a review.

North African countries; Algeria, Egypt, Libya, Morocco and Tunisia suffer from mycotoxin contamination. Various studies have indicated the presence of mycotoxins in raw milk and cereals (i.e. wheat, barley, maize and cereal-based products). Aflatoxins (AFs), Aflatoxin M1 (AFM1), Ochratoxin A (OTA), Fumonisin (FB1) and Zearalenone (ZEN)-mycotoxin are the most detected due to climatic change in the region. In this review, we will present the kind of foods and feeds cereals and milk based products contaminated and the level of their contaminated mycotoxin. On the other hand, researchers try to find biologic methods to remove/mitigate mycotoxins in food and feed using bio-products. But the research works concerning legislations and mycotoxin risk assessment still rare. Therefore, it appears necessary to make review on the current status of mycotoxins in North African countries in order to explore data related to contamination of basic food in this region and to highlight the problem to the policy-makers to establish a serious legislation on this matter. On the other hand, to give more information to the worldwide readers about the impact of climate change on the food and feed pollution on mycotoxins in the Mediterranean Sea region.

The ameliorative effect of Lactobacillus paracasei BEJ01 against FB1 induced spermatogenesis disturbance, testicular oxidative stress and histopathological damage.

Fumonisin B1 (FB1) is a possible carcinogenic molecule for humans as classified by the International Agency for Research on Cancer (IARC) in 2B group. In livestock, it is responsible for several mycotoxicoses and economic losses. Lactobacillus strains, inhabitants of a wide range of foodstuffs and the gastrointestinal tract, are generally recognized as safe (GRAS). Thus, the aim of this work was to evaluate the protective effect of Lactobacillus paracasei (LP) against FB1-induced reprotoxicities including testicular histopathology, sperm quality disturbance, and testosterone level reduction.Pubescent mice were divided randomly into four groups and treated for 10 days. Group 1: Control; Group 2: FB1 (100 µg/kg b.w); Group 3: LP (2 × 109 CFU/kg b.w); Group 4: LP (2 × 109 CFU/kg b.w) and FB1 (100 µg/kg b.w). After the end of the treatment, animals were sacrificed. Plasma, epididymis, and testis were collected for reproductive system studies.Our results showed that FB1 altered epididymal sperm quality, generated oxidative stress, and induced histological alterations. Interestingly, these deleterious effects have been counteracted by the LP administration in mice.In conclusion, LP was able to prevent FB1-reproductive system damage in BALB/c mice and could be validated as an anti-caking agent in an animal FB1-contaminated diet.

Non-competitive interactions between hydroxychloroquine and azithromycin: Systematic density functional, molecular dynamics, and docking calculations.

In this study, density functional theory (DFT) and docking calculations were systematically performed to study the non-competitive interaction between Hydroxychloroquine (HCQ) and azithromycin (AZTH). The calculated changes in Gibbs free energy and enthalpy (at 310 K) were positive, indicating the non-spontaneous formation of HCQ-AZTH specifically in water media. Docking calculation confirmed the obtained DFT result as evident from the different binding sites of both drugs to the SARS-CoV-2 main protease and human angiotensin-converting enzyme 2 (ACE2) proteins. The HCQ-AZTH structure revealed enhanced electrochemical properties, suggesting the synergy between HCQ and AZTH without affecting their therapeutic efficacy against SARS-CoV-2.

Antibacterial and anti-inflammatory activities of cardamom (Elettaria cardamomum) extracts: Potential therapeutic benefits for periodontal infections.

Cardamom (Elettaria cardamomum) is a traditional aromatic plant for which several pharmacological properties have been associated. In this study, the antibacterial activity of two cardamom extracts (fruit and seeds), rich in volatile compounds, against major periodontal pathogens was evaluated. Moreover, the ability of the extracts to exert anti-inflammatory activity was tested. Both cardamom fruit and seed extracts exerted an antibacterial effect against Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, and Prevotella intermedia (minimum inhibitory concentrations: 0.5% [v/v], 0.25%, 0.062%, 0.125%, respectively and minimum bactericidal concentrations: 1%, 0.25%, 0.062%, 0.25%, respectively). The cell membrane of P. gingivalis was disrupted by a treatment with cardamom extracts suggesting the bactericidal mode of action. The extracts also inhibited biofilm formation although it correlated with a growth reduction. Moreover, the cardamom extracts significantly decreased the secretion of IL-1ß, TNF-α, and IL-8 by lipopolysaccharide-stimulated macrophages. Evidence were brought that the anti-inflammatory activity may result from inhibition of the NF-κB signaling pathway. This study is the first to provide evidence that cardamom fruit and seed extracts through their antibacterial and anti-inflammatory properties may be therapeutic agents of interest against periodontal infections.

Analysis of a long term starved Pseudomonas aeruginosa ATCC27853 in seawater microcosms.

The persistence of pathogenic bacteria in the marine environment has been thoroughly investigated. The potential threat that these microorganisms pose to public health in recreational waters has always been a concern. In this study, the persistence and the response of Pseudomonas aeruginosa ATCC27853 to starvation and osmotic stress were studied after its incubation in sterilized seawater during 12 months. Three different colonial variants were isolated: A7 after one month, and A81 and A82 after 8 months of incubation period. The incubation effect on the bacterial phenotype and genotype were studied by analyzing modifications in morphology, antibiotic and metal resistance, molecular typing (PFGE and MLST), pigment production and virulence factors. The starved variants showed three different colony forms, but an indistinguishable PFGE pattern and belonged to ST155, as P. aeruginosa ATCC27853. The starved variants maintained the susceptibility to the 13 tested antibiotics, with the exception of the imipenem-resistant A82 strain, which also showed a small colony variant phenotype and the highest values of tolerance to the CuSO4 + NaCl combination. Significant differences were detected in the pigment production, the elastase activity and cytotoxic potential of the starved isolates in comparison to P. aeruginosa ATCC27853. Long-term exposure to stress, such as the incubation in seawater, was shown to induce different responses in P. aeruginosa, including virulent and resistant phenotypes.

Virucidal Effect of Guggulsterone Isolated from Commiphora gileadensis.

Commiphora gileadensis, locally known as becham, is a plant used in traditional Arabian medicine for treating headache, constipation, stomach, joint pain, and inflammatory disorders. Several studies have reported its antibacterial properties; however, no study has demonstrated its antiviral activity. This study aimed to evaluate the antiviral activity of C. gileadensis as well as to isolate its active compound and investigate its mode of action. This activity was evaluated using 4 viruses, herpes simplex virus type 2 (HSV-2), respiratory syncytial virus type B (RSV-B), coxsackie virus B type 3, and adenovirus type 5 by performing the plaque reduction assay and the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays for enveloped and nonenveloped viruses, respectively. The methanol extract of C. gileadensis leaves only showed antiviral activity against enveloped viruses with a selectivity index of 11.19 and 10.25 for HSV-2 and RSV-B, respectively. The study of the mechanism underlying antiviral activity demonstrated a virucidal effect by direct contact with these target viruses. The active compound, isolated using bio-guided assays involving TLC, was identified as guggulsterone by HPLC-diode array detection coupled with electrospray ionization mass spectrometry. Guggulsterone is an antagonist of the bile acid receptor and a modulator of cholesterol metabolism; however, its antimicrobial properties have been reported for the first time in this study.

Synthesis and evaluation of antibacterial and antibiofilm activities of pyridin-2-yl hexanoate.

Synthesized Pyridin-2-yl hexanoate was screened for its antibacterial and antibiofilm activities using the broth microdilution method and XTT assay respectively. The obtained results revealed that synthesized molecule inhibited the growth of pathogenic bacteria (MICs value ranged from 80 to 640 µg ml-1). Moreover, it induced a strong antibiofilm effect against Gram positive cocci (BIC50 was 366, 378 and 412 µg ml-1 for Streptococcus mutans ATCC 25175, Streptococcus salivarius ATCC 13419 and Streptococcus oralis ATCC 6249 respectively). Basis on these results, pyridin-2-yl hexanoate may be considered as effective compound with antibacterial and antibiofilm activities.

Use of carvacrol, thymol, and eugenol for biofilm eradication and resistance modifying susceptibility of Salmonella enterica serovar Typhimurium strains to nalidixic acid.

The Aims of the study was to evaluate the antibacterial susceptibility and the biofilm eradication of three natural compounds carvacrol (CAR), thymol (TH) and eugenol (EUG), alone or in combination with nalidixic acid (NA) against twelve Salmonella Typhimurium strains. The minimum inhibitory concentration (MIC) and the minimum biofilm eradication concentration (BEC50) of the tested compounds (CAR, TH and EUG) and their combinations with NA were evaluated. In order to assess whether these bacteria had active efflux pumps, ethidium bromide (EtBr) accumulation assays was achieved using spectrophotometric accumulation assays. Moreover, scanning electron microscopy was used to visualize the bacterial biofilm formation on stainless steel surfaces after exposed to NA, CAR, TH and EUG alone and in combination. TH was the most effective essential oil, with the lowest MICs values ranging from 32 to 128 µg/mL followed by EUG and CAR. In addition, the combination of NA with the different compounds enhances antibiotic susceptibility of the tested bacterial strains. These results were confirmed by EtBr accumulation assays. A pronounced effect in decreasing biofilm mass was also noticed. Moreover, SEM revealed that bacterial membrane was disrupted and a complete loss of membrane integrity was also evident. The combination of natural compounds with antibiotic enhances bacterial susceptibility to NA. This combination ameliorates eradication of biofilm formed by S. Typhimurium on polystyrene microtitre plates. Additionally, this synergy induces an alteration of the bacterial cell surface visualized by SEM.

Synergistic effect of eugenol, carvacrol, thymol, p-cymene and γ-terpinene on inhibition of drug resistance and biofilm formation of oral bacteria.

Dental caries remains the most prevalent oral infectious disease worldwide. In this study, the antibacterial and the antibiofilm activities of five essential oils (EO's): eugenol (EUG), carvacrol (CAR), thymol (TYH), p-cymene (CYM) and γ-terpinene (TER) were tested (alone or in combinaison with tetracycline) against oral bacteria. In addition, their potential roles to enhance the accumulation of ethidium bromide (EtBr) in bacterial cells were tested. Our results indicated that EO's induced a selective antimicrobial activity. A synergistic effect of EO's and tetracycline (TET) was noticed with a reduction rate ranged from 2 to 8-fold. In addition, the efflux of EtBr was inhibited with a decrease in loss of EtBr from the bacteria. On the other hand a significant anti-biofilm activities of EO's (alone or combined with antibiotics) was noticed. In conclusion the tested EO's may be considered as a potential natural source with a resistance-modifying activity and may be applied to eradicate bacterial biofilm.

Culture conditions improvement of Crassostrea gigas using a potential probiotic Bacillus sp strain.

It is well demonstrated that some probiotics improve rearing water quality and thereby have beneficial effects on reared organisms. We conducted this study to determine the effect of Bacillus consortium on Crassostrea gigas reared in contemned seawater with indigo dye priory treated with Bacillus or no treated. This effect was studied by assessing hemocytes death using flow cytometry analysis. We found that the percentage of decolorization of indigo dye in polluted seawater in presence of C. gigas increased from 41% to 90% when using Bacillus consortium. In these conditions, the hemocytes mortality of reared C. gigas decreased from 87% to 56%. We have demonstrated also that seawater contemned with priory treated indigo with Bacillus consortium is less toxic than seawater contemned with the no treated indigo. The percentage of hemocytes death is 81% for the contemned seawater with indigo and 56% for no contemned seawater. This consortium shows a protector effect of C. gigas against Vibrio harveyi contemning reared seawater.

Antibacterial and efflux pump inhibitors of thymol and carvacrol against food-borne pathogens.

In this study thymol (THY) and carvacrol (CAR), two monoterpenic phenol produced by various aromatic plants, was tested for their antibacterial and efflux pump inhibitors potencies against a panel of clinical and foodborne pathogenes. Our results demonstrated a substantial susceptibility of the tested bacteria toward THY and CAR. Especially, THY displayed a strong inhibitory activity (MIC's values ranged from 32 to 64 µg/mL) against the majority of the tested strains compared to CAR. Moreover, a significant reduction in MIC's of TET and benzalkonium chloride (QAC) were noticed when tested in combinations with THY and CAR. Their synergic effect was more significant in the case of THY which resulted a reduction of MIC's values of TET (2-8 fold) and QAC (2-8 fold). We noted also that THY and CAR inhibited the ethidium bromide (EtBr) cell efflux in a concentration-dependent manner. The rate of EtBr accumulation in food-borne pathogen was enhanced with THY and CAR (0, 250 and 500 µg/mL). The lowest concentration causing 50% of EtBr efflux inhibition (IC 50) was noticed in Salmonella enteritidis (1129) at 150 µg/mL of THY and 190 µg/mL of CAR respectively. These findings indicate that THY and CAR may serve as potential sources of efflux pump inhibitor in food-borne pathogens.

Anti-bacterial and anti-biofilm activity of probiotic bacteria against oral pathogens.

In this study, three lactic acid bacteria (LAB), isolated from barley, traditional dried meat and fermented olive were characterized and tested for their anti-bacterial and anti-biofilm activities against oral bacteria. Our results revealed that the tested LAB were γ-hemolytic and were susceptible to four antibiotics. All the strains were resistant to low pH, bile salt, pepsin and pancreatin. Furthermore, FB2 displayed a high aut-oaggregative phenotype (99.54%) while FF2 exhibited the best co-aggregation rate. Concerning the microbial adhesion to solvent, FB2 was the most hydrophobic strain (data obtained with chloroform and n-hexadecane). In addition Pediococcus pentosaceus FB2 and Lactobacillus brevis FF2 displayed a significant inhibitory effect against Streptococcus salivarius B468 (MIC = 10%). Moreover the selected strains were able to inhibit biofilm formation of Bacillus cereus ATCC14579 (MBIC50 = 28.16%) and S. salivarius B468 (MBIC50 = 42.28%). The selected LAB could be considered as candidate probiotics for further application in functional food and mainly in the prevention of oral diseases.

Molecular identification of potential denitrifying bacteria and use of D-optimal mixture experimental design for the optimization of denitrification process.

Three bacterial strains (TE1, TD3 and FB2) were isolated from date palm (degla), pistachio and barley. The presence of nitrate reductase (narG) and nitrite reductase (nirS and nirK) genes in the selected strains was detected by PCR technique. Molecular identification based on 16S rDNA sequencing method was applied to identify positive strains. In addition, the D-optimal mixture experimental design was used to optimize the optimal formulation of probiotic bacteria for denitrification process. Strains harboring denitrification genes were identified as: TE1, Agrococcus sp LN828197; TD3, Cronobacter sakazakii LN828198 and FB2, Pedicoccus pentosaceus LN828199. PCR results revealed that all strains carried the nirS gene. However only C. sakazakii LN828198 and Agrococcus sp LN828197 harbored the nirK and the narG genes respectively. Moreover, the studied bacteria were able to form biofilm on abiotic surfaces with different degree. Process optimization showed that the most significant reduction of nitrate was 100% with 14.98% of COD consumption and 5.57 mg/l nitrite accumulation. Meanwhile, the response values were optimized and showed that the most optimal combination was 78.79% of C. sakazakii LN828198 (curve value), 21.21% of P. pentosaceus LN828199 (curve value) and absence (0%) of Agrococcus sp LN828197 (curve value).

Use of juglone as antibacterial and potential efflux pump inhibitors in Staphylococcus aureus isolated from the oral cavity.

In this study the minimal inhibitory concentration (MICs) of tetracycline (Tet), erythromycin (Ery) and benzalkonium chloride (BC) in absence and in presence of a sub-MIC of juglone (Jug) were determined. In addition, the Ethidium bromide (EtBr) efflux assay was performed to assess the effect of Jug on EtBr cells accumulation. Our results showed a selective antimicrobial activity of Jug against the tested strains. A synergistic effect of Jug, drugs (Tet and Ery) and disinfectant (BC) was noticed with a reduction rate varied from 2 to 16-fold. In addition, the efflux of EtBr was inhibited depending on the Jug concentration. In the presence of Jug, a decrease in loss of EtBr from bacteria was observed. The concentration inducing 50 % of EtBr efflux inhibition after 15 min was about 182 µg ml-1 for S. aureus ATCC 25923, 236 µg ml-1 for S. aureus B193 and 195 µg ml-1 for S. aureus B456. It appears from this study that Jug may be used as a natural source for resistance-modifying activity in same bacteria.

Drug resistance of bacterial dental biofilm and the potential use of natural compounds as alternative for prevention and treatment.

Oral diseases, such as dental caries and periodontal disease are directly linked with the ability of bacteria to form biofilm. The development of dental caries involves acidogenic and aciduric Gram-positive bacteria colonizing the supragingival biofilm (Streptococcus, Lactobacillus and Actinomycetes). Periodontal diseases have been linked to anaerobic Gram-negative bacteria forming a subgingival plaque (Porphyromonas gingivalis, Actinobacillus, Prevotella and Fusobacterium). Cells embedded in biofilm are up to 1000-fold more resistant to antibiotics compared to their planctonic ones. Several mechanisms have been proposed to explain biofilms drug resistance. Given the increased bacterial resistance to antibiotics currently used in dentistry, a great importance is given to natural compounds for the prevention of oral bacterial growth, adhesion and colonization. Over the past decade, interest in drugs derived from medicinal plants has markedly increased. It has been well documented that medicinal plants and natural compounds confer considerable antibacterial activity against various microorganisms including cariogenic and periodontal pathogens. This paper provides a review of the literature focusing on the studies on (i) biofilm in the oral cavity, (ii) drug resistance of bacterial biofilm and (iii) the potential use of plant extracts, essential oils and natural compounds as biofilm preventive agents in dentistry, involving their origin and their mechanism of biofilm inhibition.

Molecular detection of bacteria associated to dental caries in 4-12-year-old Tunisian children.

The occurrence of several microbial species in the oral cavity of 4-12-year-old Tunisian children was investigated. Samples were taken from 158 children (81 caries actives and 77 caries free). Genomic DNA was extracted and analyzed for the presence of 17 microbial species using a polymerase chain reaction assay. All samples were positive for at least one of the target microbial strains. Streptococcus mutans was the most prevalent species (76.5%) detected in genomic DNA collected from carious lesions. Other prevalent species were Candida spp (63%), Streptococcus salivarius (59%) and Streptococcus oralis (42%). The frequency of Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus casei-group in caries lesions was 29.5%, 34.5% and 22% respectively. Pathogenic bacteria such as Staphylococcus aureus was found in 28.5% of carious lesion samples compared to 15.5% in the control. Frequency of Porphyromonas endodontali, Actinomyces radicidentis and Treponema denticola recovery did not differ significantly between origins of samples. PCR analysis of genomic DNA detect various oral bacteria that differ between caries actives and caries-free children. In addition, the association of same aciduric bacteria (S. mutans, S. salivarius, L. acidophilus) and caries formation was noticed.

Detection and characterization of the most common foodborne pathogens by using multiplex PCR procedure.

Food Microbial contamination is one of the most serious problems. A large percentage of food-borne illnesses are caused by food-borne pathogens, and diarrheal agents comprise more than half of the overall prevalence of food-borne illnesses in the globe, and more commonly in developing countries. This study aimed to identify the most-common foodborne organisms from foods in Khartoum state by PCR. A total of 207 food samples (raw milk, fresh cheese, yogurt, fish, sausage, mortadella, and eggs) were collected. DNA was extracted from food samples by guanidine chloride protocol, and then species-specific primers were used to identify Escherichia coli O157: H7, Listeria monocytogenes, Salmonella spp., Vibrio cholerae, V. parahaemolyticus, and Staphylococcus aureus. Out of 207 samples, five (2.41%) were positive for L. monocytogenes, one (0.48%) was positive for S. aureus, and one (0.48%) was positive for both Vibrio cholerae and Vibrio parahaemolyticus. From 91 fresh cheese samples, 2 (2.19%) were positive for L. monocytogenes, and one (1.1%) sample was positive for two different foodborne pathogens (V. cholerae and V. parahaemolyticus). Out of 43 Cow's milk samples, three (7%) samples were positive for L. monocytogenes, and out of 4 sausage samples, one (25 %) was positive for S. aureus. Our study revealed the presence of L. monocytogenes and V. cholera in raw milk and fresh cheese samples. Their presence is considered a potential problem and needs intensive hygiene efforts and standard safety measures before, during, and after food processing operations.

Whole genome sequencing analysis and Box-Behnken design for the optimization of the decolourization of mixture textile dyes by halotolerant microbial consortium.

The use of dyes in textile industries has resulted in substantially contaminated soil, water and ecosystem including fauna and flora. So, the application of eco-friendly approach for dyes removal is in great demand. The goal of this research was to develop and test a bacterial consortium for biodegrading dyes in artificial textile effluent (ATE) derived from mixture of Indigo carmine (40 mg/l); Malachite green (20 mg/l); Cotton bleu (40 mg/l); Bromocresol green (20 mg/l) and CI Reactive Red 66 (40 mg/l) dissolved in artificial seawater. The Box-Behnken design (BBD) which combine six variables with three levels each was used to determine the potential removal of dyes in ATE, by the selected microbial consortium (M31 and M69b). The experimental process indicated that decolourization of ATE reached 77.36 % under these conditions values: salinity (30 g/l), pH (9), peptone (5 g/l), inoculum size (1.5 108 CFU/ml), agitation (150 rpm) and contact time (72 h). The decolourization was confirmed by FTIR spectrum analysis of ATE before and after bacterial treatment. Bacterial strains used in this study were identified as Halomonas pacifica M31 and Shewanella algae M69b using 16 rDNA sequences. Moreover, the total genome analysis of M31 and M69b validated the implication of bacterial genes in mixture dyes removal. Therefore, the effect of the selected bacterial consortium on ATE removal was confirmed and it may be used in industrial wastewater treatment to issuing environmental safety.

In silico screening of the effectiveness of natural compounds from algae as SARS-CoV-2 inhibitors: molecular docking, ADMT profile and molecular dynamic studies.

Marine species are known as rich sources of metabolites largely involved in the pharmaceutical industry. This study aimed to evaluate in silico the effect of natural compounds identified in algae on the SARS-CoV-2 Main protease, RNA-dependent-RNA polymerase activity (RdRp), endoribonuclease (NSP15) as well as on their interaction with viral spike protein. A total of 45 natural compounds were screened for their possible interaction on SARS-CoV-2 target proteins using Maestro interface for molecular docking, molecular dynamic (MD) simulation to estimate compounds binding affinities. Among the algal compounds screened in this study, three (Laminarin, Astaxanthin and 4'-chlorostypotriol triacetate) exhibited the lowest docking energy and best interaction with SARS-CoV-2 viral proteins (Main protease, RdRp, Nsp15, and spike protein). The complex of the main protease with laminarin shows the most stable RMSD during a 150 ns MD simulation time. Which indicates their possible inhibitory activity on SARS-CoV-2.Communicated by Ramaswamy H. Sarma.

Enhanced textile dye removal from wastewater using natural biosorbent and Shewanella algae B29: Application of Box Behnken design and genomic approach.

In this study, Box-Behnken design combining seven factors at three levels were used to optimize the elimination of CI Reactive Red 66 in artificial seawater, by the combination of eco-friendly bio-sorbents and acclimated halotolerant microbial strain. Results showed that macro-algae and cuttlebone (2 %) were the best natural bio-sorbent. Additionally, the selected halotolerant strain able to rapidly remove dye was identified as Shewanella algae B29. The optimization process revealed that decolourization of CI Reactive Red 66 yields reached 91.04 % under the following variable values: dyes concentration (100 mg/l), salinity (30 g/l), peptone (2 %), pH (5), algae C (3 %), cuttlebone (1.5 %) and agitation (150 rpm). The whole genome analysis of S. algae B29 demonstrated the presence of several genes coding for valuable enzymes involved in textile dyes biotransformation, adaptation to stress as well as biofilm formation implying its potential use in biological textile wastewater treatment.