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The development of multi-drug-resistant bacteria pushed the scientific community to look for new alternatives to solve the problem. Propolis is a beehive substance and one of the richest natural products in bioactive compounds with antibacterial activity. This study was aimed to investigate the possible synergistic interaction between propolis and antibacterial drugs, such as essential oils (EOs) and antibiotics, in order to find increased activity with decreased concentrations. Two ethanol extracts of propolis were used for the test, which were collected from the north of Morocco. The chemical composition was determined by UHPLC-MS. The synergistic effect of propolis extracts with EOs and antibiotics was tested using the checkerboard technique. The chemical analysis showed the presence of more that 100 compounds in propolis extracts, belonging mainly to flavonoids. The combination of propolis with the other antibacterial drugs showed different types of interactions with FIC index values varied from 0.18 to 1, but no antagonist effect was noticed. With FICI

Camphene (C10H16) also-called2,2-dimethyl-3-methylidenebicyclo[2.2.1] heptane is a volatile compound belonging to the terpenoid family, in particular to the group of monoterpene hydrocarbons. It is...

The current study aimed to develop, optimize, and evaluate the real-time polymerase chain reaction (qPCR) specificity and sensitivity on pure isolated strains of Campylobacter and on enriched broth for direct detection of these cells. To achieve this, HipO and CadF gene were purchased to target C. jejuni and C. coli, respectively. Using chessboard titration method, three critical reagents concentration of qPCR were optimized and described. After that, 130 of collected isolates from food samples were confirmed by the developed protocols. Lastly, 27 enriched broths culture-positive for Campylobacter spp. were subjected to direct qPCR. The earlier results indicated that the diluted DNA templates improved the amplification plot intensity (∆Rn ˃ 225.000) and the amendment of the amount of specific primers, probes, as well as the concentration of MgCl2 on the amplification curves affected positively the fluorescence signal for both C. jejuni and C. coli established protocols. The confirmation of 130 of C. jejuni and C. coli isolated from broiler chickens, turkeys, cattle, and handling surface swabs by qPCR optimized protocols demonstrated similar outcomes as phenotypic tests. From 27 broiler chickens’ samples, C. coli were the most isolated strain from culture method, biochemically confirmed, compared to C. jejuni. In contrast, the application of qPCR directly on enriched broth revealed higher percentage (74.07%) of co-existence of C. coli and C. jejuni from the same analyzed samples. The present study developed and validated reliable, rapid, robust, and specific qPCR method for screening, quantifying, and confirming Campylobacter in food safety monitoring.

Recently, a new coronavirus, named Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2) has emerged in Wuhan, Hubei province, China SARS-CoV-2 caused a global epidemic (COVID-19), which continues to kill thousands of people around the world today With an exclusively interhuman transmission mode, in the absence of a vaccine, the spreading of this new coronavirus is accelerating remarkably With the studies of the molecular biology of the virus, molecular and serological diagnostic tests are implemented However, the pathophysiology is not well understood and variable, which gives different symptoms, with a significant mortality rate Indeed, several organs such as the lungs, brain, kidneys, immune system, and heart are affected directly and/or indirectly © 2020 by the authors

Campylobacter remains the leading cause responsible of human gastroenteritis worldwide. The current study aimed to compare the internal DNA extraction protocol with two commercially available kits, using C. jejuni (ATCC® 29428TM) and C. coli (ATCC®43478TM) and to validate and optimize the internal protocol through artificial contamination and confirmation of Campylobacter spp. from broiler chickens, turkeys, and beef meats samples. The extraction processes were carried out following the internal protocols steps and the manufacturer's instructions of PureLinkTM Genomic DNA Mini Kit and Wizard® Genomic DNA Purification Kit, respectively. The agarose gel electrophoresis system was used to control the DNA quality. After that, forty Campylobacter spp. isolates were confirmed, and finally, an artificial contamination of the aforementioned reference strains at three different concentrations was performed with sterilized minced meats. This is the first work comparing the PureLinkTM Genomic DNA Mini Kit and Wizard® Genomic DNA Purification Kit with internal genomic DNA extraction protocol for Campylobacter spp. The results indicated that the internal protocol provided similar efficiency to the two kits. All confirmed isolates were successfully amplified, in which28 isolates were C. coli and 12 were C. jejuni, as revealed by biochemical tests. A positive amplification was also observed in the three contaminated food matrices, after enrichment, at all examined doses. Except some reactions that were negative at 1 CFU/mL of C. jejuni and C. coli. This was explained by the detection limits of both internal protocol and qPCR. Based on our findings, three crucial steps in determining the extraction of DNA quality of this protocol were amended. Hence, the study highlighted the importance of validating simpler, cheaper and faster DNA extraction protocol, for each laboratory, as part of future risk assessment, control and monitoring programs of Campylobacter frequency required in molecular studies.

Ethnopharmacological relevance Centaurium erythraea is an important medicinal plant in many countries, e.g. Morocco, Algeria, Italy, Spain, Portugal, and countries of Balkan Peninsula. It is used in folk medicine to treat various illnesses. It is also used as an antiapoplectic, anticoagulant, anticholagogue, antipneumonic, hematocathartic, and as a hypotensive agent. Aim of the review In this review, previous reports on the taxonomy, botanical description, geographic distribution, ethnomedicinal applications, phytochemistry, pharmacological properties, and toxicity of Centaurium erythraea were critically summarized. Materials and methods Scientific search engines including PubMed, ScienceDirect, SpringerLink, Web of Science, Scopus, Wiley Online, SciFinder, and Google Scholar were consulted to collect data on C. erythraea. The data presented in this work summarized the main reports on C. erythraea phytochemical compounds, ethnomedicinal uses, and pharmacological activities. Results C. erythraea is used in traditional medicine to treat various diseases such as diabetes, fever, rhinitis, stomach ailments, urinary tract infections, dyspeptic complaints, loss of appetite, and hemorrhoids, and as diuretic. The essential oils and extracts of C. erythraea exhibited numerous biological properties such as antibacterial, antioxidant, antifungal, antileishmanial, anticancer, antidiabetic, anti-inflammatory, insecticidal, diuretic, gastroprotective, hepatoprotective, dermatoprotective, neuroprotective, and inhibitory agent for larval development. Phytochemical characterization of C. erythraea revealed the presence of several classes of secondary metabolites such as xanthonoids, terpenoids, flavonoids, phenolic acids, and fatty acids. Conclusions Ethnomedicinal studies demonstrated the use of C. erythraea for the treatment of various disorders. Pharmacological reports showed that C. erythraea especially its aerial parts and roots exhibited potent, and beneficial activities. These findings confirmed the link between the traditional medicinal use and the results of the scientific biological experiments. Considering these results, further investigation using diverse in vivo pharmacological assays are strongly recommended to validate the results of its traditional use. Toxicological tests and pharmacokinetic studies are also required to validate the safety and efficacy of C. erythraea and its bioactive contents.

Histamine poisoning is a significant public health problem. Therefore, the monitoring of histamine content in fish and fishery products is considered to be a crucial measure in the seafood industry. In the present study, a simple and rapid densitometric thin-layer chromatographic (TLC) method for histamine determination in fish samples was developed and validated. The samples were homogenized with 10% trichloroacetic acid and histamine was efficiently extracted. Then, an appropriate derivatization procedure was adopted with dansyl chloride. Once the derivatization was carried out, the samples were applied to silica gel TLC plates and developed by ascending chromatography with chloroform-triethylamine (6:4, v/v) as the mobile phase. The intensity of the histamine-dansyl derivative spots was measured by densitometry at 365 nm, and the quantitation was performed by BIO-1D image processing software. The validation of this method revealed good linearity and specificity over a concentration range from 6.25 to 100 mg/kg. Adequate precision was shown by relative standard deviations (RSD) smaller than 4.82%, accuracy ranged from &minus, 6.88% to 5.28%, and satisfactory recoveries ranging from 93% to 105% were obtained. The Limit of Detection and the Limit of Quantification were calculated at 4.4 mg/kg and 10.5 mg/kg, respectively. In addition, the effectiveness of the proposed method was assessed by the analysis of various samples, and the obtained results were confirmed with those achieved by the HPLC-UV method. Moreover, the developed method was found to be simple, cheap, and suitable for application to analyze several samples simultaneously.

The present work was carried out to understand the occurrence and antimicrobial susceptibility of Campylobacter spp., in various samples in Northern of Morocco. For this purpose, a random sampling was undertaken from butcher shops, traditional markets, and slaughterhouse. First, the research of Campylobacter was performed according to the Moroccan standard NM ISO: 10272-1 (2008). Second, the isolates were identified by biochemical tests and real time PCR. After the biochemical and molecular identification of suspected colonies, a disk diffusion method was executed to determine the sensitivity of Campylobacter spp. against 18 antibiotics. The results showed a moderate prevalence of Campylobacter species (130/466) recovered mainly on the Campylobacter blood base agar, where C. coli (108/130) were more prevalent comparable to C. jejuni (22/130) in poultry and cattle meat, raw milk, cloacal and surface swabs, and stool of patient suffering from diarrhea. The findings supported also the sensitivity of multiplex qPCR to detect Campylobacter strains compared to Moroccan standard NM ISO: 10272-1 (2008). Among our isolates, C. jejuni were the most susceptible strain toward colistin, florfenicol, gentamicin, streptomycin, and erythromycin. Nonetheless, the presence of multidrug Campylobacter resistant strains was highly observed in C. jejuni isolated, particularly, from broiler chickens toward the antibiotic classes of cephalosporin, penicillin, monobactam, quinolone, fluoroquinolone, sulfamide, as well as tetracycline. This may be due to common use of these drugs in veterinary medicine and farms as growth factor, which limits the usefulness of these molecules. Hence, the study highlights the importance of resistance profile monitoring of these pathogens in Northern of Morocco, in order to develop appropriate control measures and to reduce the emergence of multidrug-resistant strains.

Histamine level determination in fish and fish products is important for human safety, fish quality and food industry. For this reason, a rapid, robust, and precise method is needed. To achieve this objective, an HPLC method with fluorescence detection was developed and validated. Histamine in fish samples was efficiently extracted with perchloric acid, and purified with ion-exchange solid-phase extraction cartridge. A pre-column derivatization was adopted with ortho-phthalaldehyde (OPA) in the presence of the reducing agent 2-mercaptoethanol, and the stability of the histamine-OPA derivatives was achieved with the acidification of the reaction medium. In terms of validation, besides the excellent linear correlations, satisfactory recoveries at all spiking levels ranging between 0 and 200 mg kg−1 were attained, with limit of detection calculated at 1.8 mg kg−1, whereas limit of quantification determined at 5 mg kg−1. The proposed method was successfully used in the analysis of reference materials and proficiency tests, and was found to be suitable, accurate, and rapid for detection and quantification of histamine in various fish samples.

Campylobacteriosis is a zoonotic disease caused by Campylobacter, mostly associated with consumption of contaminated foodstuffs and water. Campylobacter jejuni and Campylobacter coli recognized as the leader of foodborne diarrheal illness in humans. The frequency of these microorganisms in poultry is fairly high than Salmonella and more challenging to measure, which represent an expensive burden charge on public health due to their difficulties to master them, especially with the fast increase rates of multidrug-resistant of thermophilic Campylobacter strains. It is well recognized that Campylobacter spp. is a fastidious cell, difficult to isolate in laboratories owing to their requirements and sensibility. That’s why; these factors must be taken into consideration during recovery protocols. A variety of phenotyping tests have been reported and widely used for confirmation and identification of Campylobacter species. Nonetheless, Whole Genome Sequencing (WGS) and Culture-Independent Diagnostic Tests (CIDTs) are new eras of hopeful technologies, mainly involved in the detection and characterization of threaten public health pathogens. This review aimed to describe the culture methods, phenotypic and genotypic schemes used to isolate, identify, and characterize Campylobacter isolates, through discussing the current knowledge and gaps related to the application of these techniques over others performed for typing this microaerophilic genus.

Ethnopharmacological relevance Daphne gnidium L., (Lazaz or Metnan) is a perennial plant that grows around the Mediterranean basin, in Southern Europe, North Africa and the Middle East. It is used in different countries for hair care and to treat several diseases including skin cancer, diabetes, nervous breakdowns, sinusitis, poisoning, rheumatic disorders, odontalgia, muscular pain, and gastrointestinal infections. It is also used as anti-inflammatory, insecticide, and anti-parasitic remedy. Aim of the review In this review, previous studies on D. gnidium including its botanical description, taxonomy, geographical distribution, medicinal use, phytochemistry, and pharmacological properties were critically highlighted and discussed for suggesting the exploration of this specie and its bioactive compounds in medical applications. Materials and methods Data on D. gnidium were gathered from Scientific search engines including PubMed, ScienceDirect, SpringerLink, Web of Science, Scopus, Wiley Online, SciFinder, and Google Scholar. Reports on D. gnidium written in English published before September 2020 were summarized. Results In traditional medicine, D. gnidium is used to treat diabetes, gastrointestinal infections, skin cancer, nervous breakdowns, and sinusitis. The extracts and essential oil of D. gnidium exhibited several biological properties such as antibacterial, antifungal, antiviral, antigenotoxic, hemolytic, anti-inflammatory, immunomodulatory, neuroprotective, allelopathic, and insecticidal effects. Phytochemical investigations identified several chemical classes of secondary metabolites in D. gnidium essential oil and extracts including terpenoids, coumarins, flavonoids, fatty acids, and alkanes. Conclusions The findings presented in this study showed a link between the traditional medicinal use and scientific biological results about D. gnidium. However, further investigations should be carried out to support medical and cosmetic applications of this species. Indeed, D. gnidium and its main compounds should be confirmed concerning their safety and their bioavailability. Moreover, pharmacodynamic studies should be conducted to support their efficacy in medical applications.