19 research outputs found

    Systematic review of a RAMSAR wetland and UNESCO biosphere reserve in a climate change hotspot (Ichkeul Lake, Tunisia)

    Full text link
    Tunisia\u27s Ichkeul Lake is among the most productive ecosystems in the Mediterranean, with a great regional value thanks to its diversity of habitats. It is an important overwintering area for waterfowl species. It is a RAMSAR wetland, a National Park, a UNESCO World Heritage, and a Biosphere Reserve. This review paper provides a broad overview of the climatic, hydraulic, biogeochemical features, bio-resources, and bio-productivity of the Lake. The interconnectivity between the different environmental components of the lake is presented, highlighting the main characteristics of this vital ecosystem. Its ecosystem consists of a permanent lake bordered by temporary marshes. It is connected to the Mediterranean Sea via Bizerte Lagoon under a typical semi-arid to sub-humid bio-climate with wet and dry seasons. The winter rainfall fills up the rivers and lake with freshwater that overflows into the Tinja River. In summer, high evaporation reduces the water level and allows seawater to enter the wetland from Bizerte Lagoon. The ecosystem is threatened by pollution, the damming of its main rivers, and climate change. The unsustainable water management has resulted in fundamental environmental modifications, as evidenced by the large variation in the salinity, water level, productivity of water plants, and the decline in venue and stop-overs of waterbirds. The current situation is a warning that indicates a general perturbation of the resources of this particular site and of Tunisian wetlands in general, especially that the Mediterranean region has been designated as a climate change hotspot. Accurate hydrological management is needed to boost the physical functioning of the ecosystem, and to gain deeper knowledge of the different phases of the water cycle and its relationship to other long-term environmental cycles for sustainable water management strategies in the most water-scarce region in the world

    Differentiation of stx1A gene for detection of Escherichia coli serotype O157: H7 and Shigella dysenteriae type 1 in food samples using high resolution melting curve analysis

    No full text
    Abstract Escherichia coli serotype O157: H7 and Shigella dysenteriae type 1 as the Shiga toxin‐producing bacteria cause some acute gastrointestinal and extraintestinal diseases such as hemorrhagic uremic syndrome and bloody diarrhea in human. Stx genes are the key virulence factors in these pathogens. The aim of this study was to develop HRMA assay to differentiate stx1A gene for detection of E. coli serotype O157: H7 and Sh. dysenteriae type 1 and determine the prevalence of these pathogens in food samples using this method. PCR‐HRMA assay and gold standard methods have been carried out for identification of pathogens among 135 different food samples. We found HRMA method a sensitive and specific assay (100 and 100%, respectively) for differentiation of stx1A gene, consequently, detection of these pathogens in food samples. Also, the highest prevalence of E. coli serotype O157: H7 and Sh. dysenteriae type 1 harboring stx1A gene was observed in raw milk and vegetable salad samples, respectively. HRMA as a rapid, inexpensive, sensitive and specific method is suggested to be used for differentiation of stx1A gene to detect E. coli serotype O157: H7 and Sh. dysenteriae type 1 as the key pathogens for safety evaluation of food samples

    Development of high-resolution melting (HRM) assay to differentiate the species of Shigella isolates from stool and food samples

    No full text
    Shigella species, a group of intracellular foodborne pathogens, are the main causes of bacillary dysentery and shigellosis in humans worldwide. It is essential to determine the species of Shigella in outbreaks and food safety surveillance systems. The available immunological and molecular methods for identifying Shigella species are relatively complicated, expensive and time-consuming. High resolution melting (HRM) assay is a rapid, cost-effective, and easy to perform PCR-based method that has recently been used for the differentiation of bacterial species. In this study, we designed and developed a PCR-HRM assay targeting rrsA gene to distinguish four species of 49 Shigella isolates from clinical and food samples and evaluated the sensitivity and specificity of the assay. The assay demonstrated a good analytical sensitivity with 0.01–0.1 ng of input DNA template and an analytical specificity of 100% to differentiate the Shigella species. The PCR-HRM assay also was able to identify the species of all 49 Shigella isolates from clinical and food samples correctly. Consequently, this rapid and user-friendly method demonstrated good sensitivity and specificity to differentiate species of the Shigella isolates from naturally contaminated samples and has the potential to be implemented in public health and food safety surveillance systems

    RAPD and ERIC-PCR coupled with HRM for species identification of non-dysenteriae Shigella species; as a potential alternative method

    No full text
    Species identification of Shigella isolates are so prominent for epidemiological studies and infection prevention strategies. We developed and evaluated RAPD and ERIC-PCR coupled with HRM for differentiation of non-dysenteriae Shigella species as potential alternative methods. After isolation of eighteen Shigella strains from faecal specimens collected from children under 2 years of age with diarrhea (n = 143), the species of the isolates were identified by slide agglutination assay. Also, species were identified using developed RAPD-PCR-HRM and ERIC-PCR-HRM techniques. Differentiation of the data sets was measured by principal component analysis as a dimension reduction method. Then, sensitivity and specificity of the methods were evaluated. We found RAPD-PCR-HRM method with high sensitivity and specificity (100 and 85% respectively) to identify non-dysenteriae Shigella species in clinical specimens. However, sensitivity and specificity of ERIC-PCR-HRM were evaluated 33 and 46% respectively and significantly lower than that of RAPD-PCR-HRM assay. Regardless of inherent poor reproducibility of DNA fingerprinting-based methods, RAPD-PCR-HRM assay can be considered as a potential alternative method to identify non-dysenteriae species of Shigella in clinical specimens. As we observed in the current study, HRM technique is more rapid, inexpensive, and sensitive than gel electrophoresis method to characterize PCR amplicons

    ï»żComparing PCR-generated artifacts of different polymerases for improved accuracy of DNA metabarcoding

    No full text
    Accuracy of PCR amplification is vital for obtaining reliable amplicon-sequencing results by metabarcoding. Here, we performed a comparative analysis of error profiles in the PCR products by 14 different PCR kits using a mock eukaryotic community DNA sample mimicking metabarcoding analysis. To prepare a mock eukaryotic community from the marine environment, equal amounts of plasmid DNA from 40 microalgal species were mixed and used for amplicon-sequencing by a high-throughput sequencing approach. To compare the differences in PCR kits used for this experiment, we focused on the following seven parameters: 1) Quality, 2) Chimera, 3) Blast top hit accuracy, 4) Deletion, 5) Insertion, 6) Base substitution and 7) Amplification bias amongst species. The results showed statistically significant differences (p < 0.05) for all of the seven parameters depending on the PCR kits used. These differences may result from the different DNA polymerases included in each kit, although the result can also be influenced by PCR reaction conditions. Simultaneous analysis of several parameters suggested that kits containing KOD plus Neo (TOYOBO) and HotStart Taq DNA polymerase (BiONEER, CA, US) at the annealing temperature of 65 °C displayed better results in terms of parameters associated with chimeras, top hit similarity and deletions

    Influence of Temperature on Growth and Production of Pectenotoxin-2 by a Monoclonal Culture of Dinophysis caudata

    No full text
    The effects of temperature on growth and production of Lipophilic Toxins (LT) by a monoclonal culture of Dinophysis caudata was studied. The cell density of D. caudata increased significantly with increasing temperature, and was the highest under 27, 30, and 32.5 °C. Temperature affected the average specific growth rate (”) during the exponential growth phase (EG), which increased from 15 °C to 30 °C, and then decreased at 32.5 °C. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed that this strain of D. caudata produced only pectenotoxin-2 (PTX-2) whose concentration increased significantly with incubation period, except at 32.5 °C. It was significantly different between temperatures ≀18 °C, ≄21 °C, and 32.5 °C. The cellular toxin production (CTP, pg·cell−1·day−1) showed variation with growth phase and temperature, except at 32.5 °C. The average net toxin production (Rtox) was not affected by temperature. During EG, the average specific toxin production rate (”tox) increased significantly with increase in temperature, reaching a peak of 0.66 ± 0.01 day−1 at 30 °C, and then decreased. Over the entire growth span, ”tox was significantly correlated to ”, and this correlation was most significant at 27 and 30 °C. During EG, ”tox was affected by both temperature and growth. This study shows that temperature affects growth and toxin production of this strain of D. caudata during EG. In addition, a positive correlation was found between toxin production and growth

    Essential oil composition and antioxidant capacity of carum copticum and its antibacterial effect on staphylococcus aureus, enterococcus faecalis and escherichia coli O157:H7

    No full text
    Antibacterial activity, also antioxidant capacity of Carum copticum was evaluated; in addition, a mechanism for its antibacterial action against both Gram positive and Gram negative bacteria were introduced. Presence of antimicrobial constituents (thymol, p‐cymene, γ‐terpinene) were confirmed by gas chromatography/mass spectrometry. Micro‐well dilution and disk diffusion assays were applied to assess the antibacterial activity of C. copticum against Staphylococcus aureus, Enterococcus faecalis and Escherichia coli O157:H7. The mechanism of the antibacterial activity was also evaluated by scanning electron microscopy. Total phenolic concentration of EO was 831.16 ± 3.75 mg of gallic acid equivalents per gram of oil. The minimum inhibitory concentrations of EO against S. aureus, E. faecalis and E. coli O157:H7 were 0.044%, 0.07% and 0.05%, (vol/vol) respectively. Moreover addition of C. copticum EO to minced beef at concentrations of 0.75 and 1% significantly enhanced fresh meat odor and color413CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ302763/2014-7; 190178/2013-
    corecore