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1.
Environ Technol ; : 1-13, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38619982

RESUMO

Microbial extracellular polymeric substances (EPS) have recently emerged as significant contributors in diverse biotechnological applications. Extracellular polymeric substances (EPS), produced by a Navicula salinicola strain, have been studied for potential applications in a specific heavy metal (lead (Pb II)) removal from wastewater. The optimisation of operational parameters, mainly pH, Pb and EPS concentrations, using the Box-Behnken design (BBD) was undertaken to enhance lead uptake. The higher Pb adsorption capacity reached 2211.029 mg/g. Hydroxyl, carbonyl, carboxyl, phosphoric, and sulfhydryl groups were identified quantitatively as potential sites for Pb adsorption. EPS exhibited a notable flocculation rate of 70.20% in kaolin clay at a concentration of 15 mg/L. They demonstrated an emulsifying activity greater than 88%, showcasing their versatile potential for both sedimentation processes and stabilising liquid-liquid systems. EPS could be excellent nonconventional renewable biopolymers for treating water and wastewater.

2.
Int J Biol Macromol ; 253(Pt 2): 126757, 2023 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-37678695

RESUMO

Marine algae are the most abundant resource in the marine environment and are still a promising source of bioactive compounds including hydrocolloids. This study contributes to the evaluation of the biological and biotechnological potentials of two water soluble polysaccharides, namely alginates (AHS) and fucoidan (FHS), extracted and purified from Halopteris scoparia, an abundant Tunisian brown macroalgae collected in Tunisia (Tabarka region). The total sugars, neutral monosaccharides, uronic acids, proteins, polyphenols, and sulfate groups contents were quantified for both fractions, as well as their functional groups and primary structural features by Fourier transform infrared spectroscopy, ionic and/or gas chromatography and nuclear magnetic resonance analyses. AHS and FHS showed significant anti-inflammatory (IC50 ≈ 1 mg/mL), anticoagulant (e.g., 27-61.7 for the activated partial thromboplastin time), antihyperglycemic (0.1-40 µg/mL) and anti-trypsin (IC50 ≈ 0.3-0.4 mg/mL) effects. FHS and a hydrolyzed fraction showed a very promising potential against herpes viruses (HSV-1) (IC50 < 28 µg/mL). Besides, AHS and two hydrolyzed fractions were able to stimulate the natural defenses of tomato seedlings, assessing their elicitor capacity, by increasing the activity of phenylalanine ammonia-lyase (66-422 %) but also significantly changing the polyphenol content in the leaves (121-243 %) and roots (30-104 %) of tomato plants.


Assuntos
Phaeophyceae , Scoparia , Alga Marinha , Alga Marinha/química , Água/metabolismo , Polissacarídeos/química , Phaeophyceae/química
3.
Chemosphere ; 338: 139391, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37414298

RESUMO

Mercury (Hg) is a global pollutant that affects numerous marine aquatic ecosystems. We isolated Chlorococcum dorsiventrale Ch-UB5 microalga from coastal areas of Tunisia suffering from metal pollution and analyzed its tolerance to Hg. This strain accumulated substantial amounts of Hg and was able to remove up to 95% of added metal after 24 and 72 h in axenic cultures. Mercury led to lesser biomass growth, higher cell aggregation, significant inhibition of photochemical activity, and appearance of oxidative stress and altered redox enzymatic activities, with proliferation of starch granules and neutral lipids vesicles. Such changes matched the biomolecular profile observed using Fourier Transformed Infrared spectroscopy, with remarkable spectral changes corresponding to lipids, proteins and carbohydrates. C. dorsiventrale accumulated the chloroplastic heat shock protein HSP70B and the autophagy-related ATG8 protein, probably to counteract the toxic effects of Hg. However, long-term treatments (72 h) usually resulted in poorer physiological and metabolic responses, associated with acute stress. C. dorsiventrale has potential use for Hg phycoremediation in marine ecosystems, with the ability to accumulating energetic reserves that could be used for biofuel production, supporting the notion of using of C. dorsiventrale for sustainable green chemistry in parallel to metal removal.


Assuntos
Mercúrio , Microalgas , Mercúrio/metabolismo , Microalgas/metabolismo , Ecossistema , Proteínas de Choque Térmico , Lipídeos
4.
Environ Technol ; : 1-12, 2023 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-36952278

RESUMO

Algae constitute a significant part of marine biodiversity. They represent a renewable source of bioactive metabolites from drug development and therapeutic fields. Fucoxanthin and ß-carotene from the brown macroalgae Halopteris scoparia, were extracted using conventional organic solvent extraction, then purified, to homogeneity, based on various chromatographic principles. Their effects on digestive enzymes and harmful bacteria were investigated. The capacities of both purified pigments to inhibit α-amylase and trypsin enzymes were evaluated. Purified fucoxanthin and ß-carotene exhibited interesting α-amylase inhibition activities, with IC50 of 300 and 500 µg/mL, respectively. Moreover, trypsin inhibition activities were detected using purified these two pigments. The antibacterial potential of the purified pigments was evaluated. ß-carotene showed to be a great antibacterial natural compound against gram-positive and gram-negative bacteria such as Listeria monocytogenes, Staphylococcus aureus and Salmonella enterica with Minimal Inhibitory Concentration (MIC) of about 0.225, 0.1125, 0.225 µg/mL, respectively. Those findings are in favor of the exploitation of H. scoparia pigments in therapeutic fields as an antidiabetic source directly by the inhibition of α-amylase and trypsin as well as antibacterial agents against gastrointestinal infections.

5.
Anal Biochem ; 667: 115080, 2023 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-36775111

RESUMO

Parachlamydia acanthamoebae and Simkania negevensis, two Chlamydia-like bacteria, have been recently recognized as emerging human respiratory pathogens. The prevalence and frequency of these bacteria in the environment and among atypical pneumonia patients are still underestimated by classical cultures, immunohistochemistry and serology which are non-specific, long and tedious methods. This study aims to develop a new duplex probe-based q-PCR assay for the simultaneous detection and quantification of P. acanthamoebae and S. negevensis. The selected hydrolysis probes displayed no cross-reaction with the closely related Chlamydia or the other tested waterborne pathogens. The assay achieved a large dynamic range for quantification (from 5 × 106 to 5 DNA copies/reaction). Efficiencies of FAM and JOE label probes weren't affected when they were combined. They were close to 100%, indicating the linear amplification. The application of this diagnostic tool resulted in 9/47 (19%) and 4/47 (8.5%) positive water samples for P. acanthamoebae and S. negevensis, respectively. P. acanthamoebae was also covered from 2/78 (2.5%) respiratory specimens and only one case (1/200 = 0.5%) of P. acanthamoebae and SARS-CoV-2 co-infection was noticed. While S. negevensis wasn't detected in clinical samples, the developed duplex q-PCR was shown to be an accurate, highly sensitive, and robust diagnostic tool for the detection and quantification of P. acanthamoebae and S. negevensis.


Assuntos
COVID-19 , Humanos , SARS-CoV-2 , Reação em Cadeia da Polimerase/métodos , Teste para COVID-19
6.
Environ Sci Pollut Res Int ; 30(7): 17564-17572, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36195814

RESUMO

Human respiratory infections caused by a large variety of microbial pathogens are the most common diseases responsible for hospitalization, morbidity and mortality. Parachlamydia acanthamoebae, a Chlamydia-related bacterium, has been found to be potentially associated with these diseases. An early and accurate diagnosis of this pathogen could be useful to avoid the potential respiratory complications linked especially to COVID-19 patients and to set suitable outbreak control measures. A TaqMan-PCR assay was developed to detect and quantify Parachlamydia acanthamoebae in environmental and clinical samples from patients of all ages with COVID-19. The selected hydrolysis probe displayed no cross-reaction with the closely related Chlamydia or the other tested pathogens. This q-PCR achieved good reproducibility and repeatability with a detection limit of about 5 DNA copies per reaction. Using this q-PCR assay, Parachlamydia acanthamoebae was detected in 2/78 respiratory specimens and 9/47 water samples. Only one case (1.3%) of Parachlamydia acanthamoebae and SARS-COV-2 co-infection was noticed. To our knowledge, the combination of these two respiratory pathogens has not been described yet. This new TaqMan-PCR assay represents an efficient diagnostic tool to survey Parachlamydia acanthamoebae on a large-scale screening programs and also during outbreaks.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , SARS-CoV-2/genética , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Teste para COVID-19
7.
Biomolecules ; 12(7)2022 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-35883441

RESUMO

Microalgae are photoautotrophic microorganisms known as producers of a large variety of metabolites. The taxonomic diversity of these microorganisms has been poorly explored. In this study, a newly isolated strain was identified based on the 18S rRNA encoding gene. The phylogenetic analysis showed that the isolated strain was affiliated with the Rhodomonas genus. This genus has greatly attracted scientific attention according to its capacity to produce a large variety of metabolites, including phycoerythrin. Growth and phycoerythrin production conditions were optimized using a Plackett-Burman design and response surface methodology. An expression profile analysis of the cpeB gene, encoding the beta subunit of phycoerythrin, was performed by qRT-PCR under standard and optimized culture conditions. The optimization process showed that maximum cell abundance was achieved under the following conditions: CaCl2 = 2.1328 g/L, metal solution = 1 mL/L, pH = 7 and light intensity = 145 µmol photons/m2/s, whereas maximum phycoerythrin production level occurred when CaCl2 = 1.8467 g/L, metal solution = 1 mL/L, pH = 7 and light intensity = 157 µmol/m2/s. In agreement, positive transcriptional regulation of the cpeB gene was demonstrated using qRT-PCR. This study showed the successful optimization of abiotic conditions for highest growth and phycoerythrin production, making Rhodomonas sp. suitable for several biotechnological applications.


Assuntos
Microalgas , Ficoeritrina , Biomassa , Cloreto de Cálcio/metabolismo , Microalgas/metabolismo , Filogenia
8.
Environ Sci Pollut Res Int ; 29(59): 88699-88709, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35836051

RESUMO

Karlodinium veneficum is a toxic benthic globally distributed dinoflagellate which has direct impacts on human health and the environment. Early and accurate detection of this harmful algal bloom-forming species could be useful for potential risks monitoring and management. In the present work, a real-time PCR targeting the internal transcribed spacer ribosomal DNA region for the specific detection and absolute quantification of K. veneficum was designed. Then, the assay conditions were adjusted and validated. The developed qPCR was highly specific for the target species and displayed no cross-reactivity with closely related dinoflagellates and/or other microalgal species commonly distributed along the Tunisian coast. Its lowest detection limit was 5 rDNA copies per reaction, which is often considered satisfying. qPCR assay enumeration accuracy was evaluated using artificially inoculated environmental samples. The comparison of the cell abundance estimates obtained by qPCR assay with the theoretical estimates showed no statistically significant difference across a range of concentrations. We suggest that the qPCR approach developed in the present study may be a valuable tool to investigate the distribution and seasonal dynamics of K. veneficum in marine environments.


Assuntos
Dinoflagellida , Microalgas , Humanos , Proliferação Nociva de Algas , Reação em Cadeia da Polimerase em Tempo Real , DNA Ribossômico
9.
Plant Physiol Biochem ; 186: 99-106, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-35835079

RESUMO

Given the increasing interest that is being paid to polysaccharides derived from algae as plant natural defense stimulators, the degree of sulfation of exopolysaccharides produced by P. sordidum for inducing defense responses in date palm vitroplants was investigated. Firstly, the culture parameters of P. sordidum were optimized to maximize the amount of sulfate in EPS using a Box-Behnken experimental design and the elicitor effects of two EPS which differ in the sulfation degrees were compared. Results demonstrated that the concentrations of NaCl, NaNO3 and MgSO4 set at 28, 0.54 and 16.31 g/L, respectively yielded the best sulfate contents. To elucidate defense-inducing activities in date palm vitroplants, EPS with the highest sulfate content (EPS1) were prepared for comparison with those obtained under standard conditions (EPS0). A fucoidan extracted from Cystoseira compressa was used as positive control and MgSO4 as negative control. Both EPS and the fucoidan displayed H2O2 accumulation and expression of PR1, SOD, PAL and WRKY genes. Interestingly, EPS1 was significantly more bioactive than EPS0 and the fucoidan suggesting that the elicitor activity is positively correlated with the sulfate groups content of this polysaccharide.


Assuntos
Phaeophyceae , Phoeniceae , Porphyridium , Peróxido de Hidrogênio , Polissacarídeos , Sulfatos/farmacologia
10.
Environ Sci Pollut Res Int ; 29(42): 63953-63963, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35469376

RESUMO

Gymnodinium catenatum is a dinoflagellate known to cause paralytic shellfish poisoning (PSP), commonly associated with human muscular paralysis, neurological symptoms, and, in extreme cases, death. In the present work, we developed a real-time PCR-based assay for the rapid detection of the toxic microalgal species, G. catenatum, in environmental bivalve mollusc samples as well as seawater samples. G. catenatum-specific primers and probe were designed on the ITS1-5.8S-ITS2 rDNA region. Hydrolysis probe qPCR assay was optimized. ITS1-5.8S-ITS2 rDNA region copy numbers per G. catenatum cell genome were estimated to be 122.73 ± 5.54 copies/cell, allowing cell quantification. The application of the optimized qPCR assay for G. catenatum detection and quantification in field samples has been conducted, revealing high sensitivity (detection of around 1.3105 cells/L of seawater samples. Thus, the designed hydrolysis probe qPCR assay could be considered an efficient tool for phytoplankton monitoring whilst ensuring accuracy and sensitivity and providing cost and time savings.


Assuntos
Dinoflagellida , Proliferação Nociva de Algas , DNA Ribossômico/genética , Reação em Cadeia da Polimerase em Tempo Real , Tunísia
11.
Sci Total Environ ; 799: 149464, 2021 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-34388883

RESUMO

Metal elements are widely used in various industrial activities and are considered as common water source contaminants. Thus, the development of cost-effective, simple design and efficient processes for trace metal elements removal from contaminated water sources is of great interest. The effects of cadmium, lead and chromium on growth, biomolecules accumulation and metabolic responses of Amphora coffaeiformis, Navicula salinicola and Dunaliella salina isolated from Tunisian coasts were tested. The bioremediation capacities of the three microalgae strains and the mechanisms involved in ions metal removal were also investigated. N. salinicola and D. salina seem to be better tolerating to Cr, while A. coffaeiformis and N. salinicola showed high resistance to Pb. The expression profile analyses by qRT-PCR of the antioxidant defense-related genes revealed that Cd, Pb and Cr treatments induce the up-regulation of catalase and superoxide dismutase coding genes for A. coffaeiformis and D. salina. Regarding N. salinicola, the catalase coding gene seems to be overexpressed after Cd, Pb and Cr exposure while only Cd and Cr induce superoxide dismutase gene overexpression. Moreover, the phytochelatin synthase (a metal chelator synthesis-related gene) was up-regulated in N. salinicola, A. coffaeiformis and D. salina after Cr exposure and also in A. coffaeiformis and D. salina after Cd exposure. While Pb treatments induce overexpression of phytochelatin synthase coding gene only for D. salina. Studied strains showed promising metal removal efficiencies for both Pb and Cr ions metals reached 95% for D. salina. Ion metal removal mechanisms study revealed that intracellular bioaccumulation process is used by D. salina for Cr up-taking. However, both intracellular and extracellular removal mechanisms are involved for Pb and Cr removal using A. coffaeiformis, N. salinicola and for Pb removal using D. salina. FTIR analysis demonstrated that several functional groups as carboxyl, hydroxyl, amino, phosphate and sulfate may participate in the bioadsorption process.


Assuntos
Metais Pesados , Microalgas , Poluentes Químicos da Água , Cádmio , Cromo/análise , Chumbo , Poluentes Químicos da Água/análise
12.
Environ Sci Pollut Res Int ; 28(41): 57486-57498, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34089447

RESUMO

Karenia selliformis is a marine dinoflagellate responsible for fish-kill events. Its presence has been reported along the Tunisian coasts (south-eastern Mediterranean Sea) since the 1990s. In the present study, a quantitative-PCR assay, based on the internal transcribed spacer (ITS) molecular marker, was developed to detect and quantify K. selliformis in environmental bivalve mollusk samples and in seawater samples. The assay was optimized, and its specificity was confirmed using cross-reactivity experiments against microalgal species commonly found on the Tunisian coasts and/or closely related to K. selliformis. Calibration curves were performed by tenfold dilutions of plasmid DNA harboring target sequence and genomic DNA, attaining a limit of detection of around 5 copies of target DNA per reaction, far below one K. selliformis cell per reaction. The field application of the developed assay showed a powerful detection capability. Thus, the designed assay could contribute to the deployment of in-field diagnostic tools for K. selliformis blooms monitoring.


Assuntos
Bivalves , Dinoflagellida , Animais , DNA , Dinoflagellida/genética , Reação em Cadeia da Polimerase em Tempo Real , Água do Mar
13.
Biomolecules ; 11(2)2021 02 14.
Artigo em Inglês | MEDLINE | ID: mdl-33672873

RESUMO

Polysaccharides from marine algae are one novel source of plant defense elicitors for alternative and eco-friendly plant protection against phytopathogens. The effect of exopolysaccharides (EPS) produced by Porphyridium sordidum on elicitation of Arabidopsis thaliana defense responses against Fusarium oxysporum was evaluated. Firstly, in order to enhance EPS production, a Box-Behnken experimental design was carried out to optimize NaCl, NaNO3 and MgSO4 concentrations in the culture medium of microalgae. A maximum EPS production (2.45 g/L) higher than that of the control (0.7 g/L) was observed for 41.62 g/L NaCl, 0.63 g/L NaNO3 and 7.2 g/L MgSO4 concentrations. Structurally, the EPS contained mainly galactose, xylose and glucose. Secondly, the elicitor effect of EPS was evaluated by investigating the plant defense-related signaling pathways that include activation of Salicylic or Jasmonic Acid-dependent pathway genes. A solution of 2 mg/mL of EPS has led to the control of fungal growth by the plant. Results showed that EPS foliar application induced phenylalaline ammonia lyase and H2O2 accumulation. Expression profile analysis of the defense-related genes using qRT-PCR revealed the up-regulation of Superoxide dismutases (SOD), Peroxidase (POD), Pathogenesis-related protein 1 (PR-1) and Cytochrome P450 monooxyge-nase (CYP), while Catalase (CAT) and Plant defensin 1.2 (PDF1.2) were not induced. Results suggest that EPS may induce the elicitation of A. thaliana's defense response against F. oxysporum, activating the Salicylic Acid pathway.


Assuntos
Arabidopsis/efeitos dos fármacos , Fusarium/imunologia , Polissacarídeos/biossíntese , Porphyridium/metabolismo , Arabidopsis/imunologia , Arabidopsis/microbiologia , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia por Troca Iônica/métodos , DNA Ribossômico/genética , Interações Hospedeiro-Patógeno , Peróxido de Hidrogênio/metabolismo , Polissacarídeos/farmacologia , Porphyridium/classificação , Porphyridium/genética , RNA Ribossômico 18S/genética
14.
Sci Total Environ ; 773: 145024, 2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-33582349

RESUMO

Phycoremediation is being considered as an eco-friendly and safe technology for toxics eradication from contaminated aquatic systems. The zinc biosorption capacity of Dunaliella sp. AL-1 was demonstrated. Zinc impacted cell growth and photosynthetic pigments accumulation showing exposure time and concentration-dependent effects. The investigation of the antioxidant protective response to zinc exposition proved a stimulation of guaiacol peroxidase (GPX) activity and an increased rate of total phenolics, flavonoids, condensed tannins and glutathione (GSH). The Box-Behnken design was used to optimize zinc removal conditions by Dunaliella sp. AL-1 strain. The maximum experimental zinc uptake was obtained when zinc concentration, algae dose, initial pH, and contact time were set at 25 mg/L, 0.5 g/L, 7.59 and 13 h 43 min, respectively. Under completely optimized conditions, the fraction of zinc removed intracellularly was much lower than the adsorbed on the cell surface. FTIR analysis Dunaliella sp. AL-1 biomass demonstrated that several functional groups as OH, CH2, CO, PO, COO and CO may participate in the biosorption process. A comparative proteomic analysis through nano-HPLC coupled to LC-MS/MS, was performed from pre- and post-zinc treatments cells. Among 199 identified proteins, 60 were differentially expressed of which 41 proteins were down-regulated against 19 up-regulated ones. Target proteins have been demonstrated to be implicated in different metabolic processes mainly photosynthesis and antioxidant defenses.


Assuntos
Efrina-A5 , Zinco , Adsorção , Biomassa , Cromatografia Líquida , Concentração de Íons de Hidrogênio , Proteômica , Espectrometria de Massas em Tandem
15.
Int J Biol Macromol ; 167: 1491-1498, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33202265

RESUMO

Proteases are industrially important catalysts. They belong to a complex family of enzymes that perform highly focused proteolysis functions. Given their potential use, there has been renewed interest in the discovery of proteases with novel properties and a constant thrust to optimize the enzyme production. In the present study, a novel extracellular neutral protease produced from Arthrospira platensis was detected and characterized. Its proteolytic activity was strongly activated by ß-mercaptoethanol, 5,5-dithio-bis-(2-nitrobenzoic acid) and highly inhibited by Hg2+ and Zn2+ metal ions which support the fact that the studied protease belongs to the cysteine protease family. Using statistical modelling methodology, the logistic model has been selected to predict A. platensis growth-kinetic values. The optimal culture conditions for neutral protease production were found using Box-Behnken Design. The maximum experimental protease activities (159.79 U/mL) was achieved after 13 days of culture in an optimized Zarrouk medium containing 0.625 g/L NaCl, 0.625 g/L K2HPO4 and set on 9.5 initial pH. The extracellular protease of A. platensis can easily be used in the food industry for its important activity at neutral pH and its low production cost since it is a valuation of the residual culture medium after biomass recovery.


Assuntos
Peptídeo Hidrolases/isolamento & purificação , Peptídeo Hidrolases/metabolismo , Inibidores de Proteases/toxicidade , Spirulina/enzimologia , Análise de Variância , Biomassa , Cisteína Proteases/metabolismo , Concentração de Íons de Hidrogênio , Íons/toxicidade , Mercaptoetanol/farmacologia , Mercúrio/toxicidade , Modelos Estatísticos , Nitrobenzoatos/farmacologia , Proteólise , Spirulina/crescimento & desenvolvimento , Zinco/toxicidade
16.
J Microbiol Methods ; 178: 106081, 2020 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-33035573

RESUMO

Prorocentrum lima (P. lima) is a widely spread dinoflagellate in the Mediterranean Sea and it has become increasingly involved in harmful algal blooms. The purpose of this study is to develop a probe-based real-time polymerase chain reaction (PCR) targeting the ITS1-5.8S-ITS2 region for the detection and absolute quantification of P. lima based on linear and circular DNA standards. The results have shown that the quantitative PCR (q-PCR), using circular plasmid as a template, gave a threshold cycle number 1.79-5.6 greater than equimolar linear standards. When microalgae, commonly found in aquatic samples were tested, no cross-amplification was observed. The q-PCR brought about a good intra and inter-run reproducibility and a detection limit of 5 copies of linear plasmid per reaction. A quantitative relationship between the cell numbers and their corresponding plasmid copy numbers was attained. Afterwards, the effectiveness of the developed protocol was tested with 130 aquatic samples taken from 19 Tunisian sampling sites. The developed q-PCR had a detection sensitivity of up to 1 cell. All the positive samples were taken from three sampling sites of Medenine Governorate with cell abundances that ranged from 22 to 156,000 cells L-1 of seawater. The q-PCR assay revealed a high sensitivity in monitoring the aquatic samples in which the low concentrations of P. lima were not accurately detected by light microscopy. Indeed, this approach is at the same time rapid, specific and sensitive than the traditional microscopy techniques and it represents a great potential for the monitoring of P. lima blooms.

17.
Mol Cell Probes ; 53: 101645, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32745685

RESUMO

Simkania negevensis is an emerging Chlamydia-like bacterium related to human respiratory diseases. An early and accurate detection of this pathogen could be useful to monitor the potential infectious risks and to set suitable outbreak control measures. In Tunisia, distribution and abundance of S. negevensis remain until now largely unknown. In the present work, a qPCR assay, targeting the 16S rRNA gene, for fast detection and quantification of S. negevensis was developed and validated. A high specificity for S. negevensis detection displaying no cross-reaction with the closely related Chlamydia spp. or the other tested microorganisms was noticed. qPCR assay performance was considered very satisfying with detection limits of 5 DNA copies per reaction. qPCR assay validation was performed by screening 37 clinical specimens and 35 water samples. S. negevensis wasn't detected in respiratory samples, but it was found in four cases of water samples. We suggest that the qPCR assay developed in this study could be considered sufficiently characterized to initiate the quantification of S. negevensis in environmental samples.


Assuntos
Chlamydiales/isolamento & purificação , Infecções por Bactérias Gram-Negativas/diagnóstico , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/genética , Chlamydiales/classificação , Chlamydiales/genética , DNA Bacteriano/genética , DNA Ribossômico/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Limite de Detecção , Sensibilidade e Especificidade , Tunísia
18.
Environ Sci Pollut Res Int ; 27(29): 36889-36899, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32577959

RESUMO

Quantitative PCR (qPCR) is the method of choice for specific detection and quantification of harmful algal bloom (HAB) species. Development of qPCR assay for simultaneous enumeration of species that frequently co-exist in HABs is required. A high sensitivity TaqMan qPCR assay, using probe and primers, located at ITS1-5.8S-ITS2 rDNA region, detecting, specifically, Karenia selliformis, K. bidigitata, and K. mikimotoi, was designed. ITS1-5.8S-ITS2 rDNA region copy numbers per Karenia cell genome were estimated to 217.697 ± 67.904, allowing cell quantification. An application of the designed methodology in field samples has been conducted, and it showed high sensitivity (detection of around 10-1 cell/100 mg of bivalve mollusk tissue, equivalent to about 20 copies of the target sequence). We suggest that the optimized method could contribute to early detection of three closely related Karenia species in seafood cultivating areas to promote control quality, guarantee a fast and effective intervention, and improve public health prevention.


Assuntos
Dinoflagellida , Primers do DNA , DNA Ribossômico , Proliferação Nociva de Algas , Reação em Cadeia da Polimerase em Tempo Real
19.
Int J Biol Macromol ; 123: 42-49, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30391590

RESUMO

Cyt1A98 is a novel cytolytic protein, from BUPM98 Bacillus thuringiensis strain, characterized by its synergistic activity with B. thuringiensis kurstaki toxins against lepidopteran larvae. In this study, we evidenced that Cyt1A98 improves the toxicity of B. thuringiensis kurstaki toxins against Aedes aegypti larvae. In fact, the strain BNS3pHTcyt1A98 exhibited a larvicidal activity of about 849-fold of that of BNS3pHTBlue against A. aegypti. The molecular and biochemical characterizations, of cyt1A98 gene and its product, were achieved. Cyt1A98 had an LC50 value of about 126.56 mg l-1 against A. aegypti larvae. Compared to Cyt1Aa of B. thuringiensis israelensis, Cyt1A98 amino acid sequence harbours three substitutions of three conserved amino acids among Cyt1Aa family members (Ser42Pro, Pro82Ala, Met188Thr). The Cyt1A98 protein structural analysis evidenced more flexibility than Cyt1Aa. According to the high fluctuation observed for the residue Pro42, the amino acid at position 42 is implicated in the flexibility property of Cyt1Aa especially for the αC and αD helices, involved in the penetration into the cell membrane. The toxicity improvement could be probably due to the higher flexibility combined with the specific affinity toward dipteran larvae. The Cyt1A/B. thuringiensis kurstaki Cry toxins model provides a potential molecular genetic strategy for an efficient bioinsecticide.


Assuntos
Bacillus thuringiensis/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/farmacologia , Toxinas Bacterianas/farmacologia , Endotoxinas/farmacologia , Proteínas Hemolisinas/metabolismo , Larva/efeitos dos fármacos , Aedes/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Controle Biológico de Vetores/métodos , Sensibilidade e Especificidade , Alinhamento de Sequência
20.
Int J Biol Macromol ; 120(Pt B): 2106-2114, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30201560

RESUMO

The production of B-phycoerythrin (B-PE) from the red microalga Porphyridium marinum was optimized before to purify it and subsequently study its antioxidant activities. NaNO3, K2HPO4 and metal traces concentrations of the culture medium, and luminosity parameters were chosen, according to the Plackett-Burman design, as the most influent factors on the B-PE production by P. marinum. The optimization of these factors according to the Box-Behnken plan gave a maximum of B-PE production equal to 40 mg/g dry weight under the following conditions: NaNO3 = 3.4 g/L; K2HPO4 = 0 g/L; light intensity = 70 µmol photons/m2/s and metal solution = 1.5 mL/L. The B-PE also showed an interesting capacity to chelate Fe3+ (IC50 = 13.91 ±â€¯0.21 µg/mL) and a significant reducing power (OD700nm = 0.485 ±â€¯0.011 at 100 µg/mL). The present study reports the antioxidant potential of purified B-PE from P. marinum that could be potentially used as a source of bioactive protein for a wide range of cosmetic and pharmaceutical applications.


Assuntos
Antioxidantes/metabolismo , Biotecnologia/métodos , Indústrias , Microalgas/metabolismo , Ficoeritrina/biossíntese , Porphyridium/metabolismo
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