Evaluating aflatoxins and Sudan dyes contamination in red chili and turmeric and its health impacts on consumer safety of Lahore, Pakistan.

Spices are contaminated with aflatoxins (AFs) and Sudan dyes which are classified as class Group 1 and Group 3 human carcinogens by the International Agency for Research on Cancer (IARC) respectively and their prolonged exposure may raise a human health concern. A total of 474 samples of red chili and turmeric were collected from Lahore city and were subjected to quantitative and qualitative AFs and Sudan dyes analysis by thin layer chromatography (TLC) respectively. The number of red chili and turmeric samples with ≥10 µg/kg of total AFs (European Union standard limit) were 70% and 33% respectively and considered unfit for human consumption. The presence of Sudan dyes in red chili and turmeric samples was 67% and 27% respectively. The mean estimated daily intake (EDI) among females and males was 0.0019 µg/kg bw/day, 0.0012 µg/kg bw/day for red chili, and 0.0008 µg/kg bw/day, 0.0006 µg/kg bw/day for turmeric respectively. The mean value of margin of exposure (MOE) among females and males for ingestion of AFs-contaminated red chili and turmeric was 210.25, 332.13, 501.02, and 699.31 respectively. Therefore, the current study demands a continuous monitoring plan and the implementation of novel techniques to enhance the product's quality and protect public health.

Isolation and Characterization of Lytic Pseudomonas aeruginosa Bacteriophages Isolated from Sewage Samples from Tunisia.

Bacteriophages could be a useful adjunct to antibiotics for the treatment of multidrug-resistant Pseudomonas aeruginosa infections. In this study, lytic P. aeruginosa myoviruses PsCh, PsIn, Ps25, and Ps12on-D were isolated from Tunisian sewage samples. Phage Ps12on-D displayed an adsorption time of ~10 min, a short latency period (~10 min), and a large burst size (~115 PFU per infected cell) under standard growth conditions. All phages were active at broad temperature (4 °C to 50 °C) and pH (3.0 to 11.0) ranges and were able to lyse a wide variety of P. aeruginosa strains isolated from clinical and environmental samples worldwide. Illumina sequencing revealed double-stranded DNA genomes ranging from 87,887 and 92,710 bp with high sequence identity to Pseudomonas phage PAK_P1. All four phages based on sequence analysis were assigned to the Pakpunavirus genus. The presented characterization and preclinical assessment are part of an effort to establish phage therapy treatment as an alternative strategy for the management of multidrug-resistant P. aeruginosa infections in Tunisia.

The exposure to polyvinyl chloride microplastics and chrysene induces multiple changes in the structure and functionality of marine meiobenthic communities.

The effects of microplastics and sorbed polycyclic aromatic hydrocarbons at community levels were rarely assessed in laboratory experiments, despite their obvious advantage in reflecting better the natural conditions compared to traditionally single species-focused toxicological experiments. In the current study, the multifaceted effects of polyvinyl chloride and chrysene, acting alone or combined, on general marine meiobenthos, but with a special focus on free-living marine nematode communities were tested in a laboratory experiment carried in microcosms. The meiobenthos was exposed to two polyvinyl chloride (5 and 10 mg.kg-1 Dry Weight 'DW') and chrysene (37.5 and 75 ng.g-1 DW) concentrations, respectively, as well as to a mixture of both compounds, for 30 days. The results highlighted a significant decrease in the abundance of all meiobenthic generic groups, including nematodes, directly with increasing dosages of these compounds when added alone. The addition of chrysene adheres to microplastics, making the sediment matrix glueyer, hence inducing greater mortality among generic meiobenthic groups. Moreover, the nematofauna went through a strong restructuring phase following the exposure to both compounds when added alone, leading to the disappearance of sensitive nematodes and their replacement with tolerant taxa. However, the similarity in nematofauna composition between control and polyvinyl chloride and chrysene mixtures suggests that the toxicity of the latter could be attenuated by its physical bonding to the former pollutant. Other changes in the functional traits within the nematode communities were a decline in the fertility of females and an increase of the pharyngeal pumping power following exposure to both pollutants for the dominant species. The latter results were also supported by additional toxicokinetics analyses and in silico modeling.

Glyphosate exposure modulates lipid composition, histo-architecture and oxidative stress status and induces neurotoxicity in the smooth scallop Flexopecten glaber.

Glyphosate is the most sprayed pesticide across the globe. Its toxicity to non-target marine organisms has recently piqued the scientific community's interest. Therefore, the purpose of this study is to investigate the potentially toxic effects of glyphosate on scallops, an ecologically and economically important bivalve group. To do that, specimens of the smooth scallop Flexopecten glaber were exposed to different concentrations (10, 100, and 1000 µg L-1) of the technical-grade glyphosate acid (GLY) for 96 h. The detrimental effects of this pollutant were assayed at cellular and tissular levels. The obtained results showed that the GLY was able to induce oxidative stress in the gills and the digestive gland of F. glaber as revealed by the enhanced hydrogen peroxide (H2O2), protein carbonyls (PCO), malondialdehyde (MDA), and lipid peroxides (LOOH) levels and the altered antioxidant defense system (the glutathione GSH content and the superoxide dismutase (SOD) activity). Additionally, GLY was found to alter the fatty acid profile, to exert a neurotoxic effect through the inhibition of the acetylcholinesterase (AChE) activity, and to provoke several histopathological damages in the two organs studied. The obtained results revealed that the pure form of GLY may exert toxic effects on F. glaber even at relatively low concentrations.

Biopriming of Durum Wheat Seeds with Endophytic Diazotrophic Bacteria Enhances Tolerance to Fusarium Head Blight and Salinity.

There is growing interest in the use of bio inoculants based on plant growth-promoting bacteria (PGPB) to promote plant growth under biotic and abiotic stresses. Here, we provided a detailed account of the effectiveness of a number of endophytic PGPB strains, isolated from the roots of the halophyte Salicornia brachiata in promoting durum wheat growth and enhancing its tolerance to salinity and fusarium head blight (FHB) disease. Bacillus spp. strains MA9, MA14, MA17, and MA19 were found to have PGPB characteristics as they produced indole-3-acetic acid, siderophores, and lytic enzymes, fixed free atmospheric nitrogen, and solubilized inorganic phosphate in vitro. Additionally, the in vivo study that involved in planta inoculation assays under control and stress conditions indicated that all PGPB strains significantly (p < 0.05) increased the total plant length, dry weight, root area, seed weight, and nitrogen, protein, and mineral contents. Particularly, the MA17 strain showed a superior performance since it was the most efficient in reducing disease incidence in wheat explants by 64.5%, in addition to having the strongest plant growth promotion activity under salt stress. Both in vitro and in vivo assays showed that MA9, MA14, MA17, and MA19 strains were able to play significant PGPB roles. However, biopriming with Bacillus subtilis MA17 offered the highest plant growth promotion and salinity tolerance, and bioprotection against FHB. Hence, it would be worth testing the MA17 strain under field conditions as a step towards its commercial production. Moreover, the strain could be further assessed for its plausible role in bioprotection and growth promotion in other crop plants. Thus, it was believed that the strain has the potential to significantly contribute to wheat production in arid and semi-arid regions, especially the salt-affected Middle Eastern Region, in addition to its potential role in improving wheat production under biotic and abiotic stresses in other parts of the world.

Molecular interactions of polyvinyl chloride microplastics and beta-blockers (Diltiazem and Bisoprolol) and their effects on marine meiofauna: Combined in vivo and modeling study.

The ecotoxicological effects of beta-blockers (i.e. Diltiazem and Bisoprolol) and their interactions with the microplastic polyvinyl chloride on marine meiofauna were tested in laboratory microcosms. An experimental factorial design was applied, using meiobenthic fauna collected from the Old Harbor of Bizerte (NE Tunisia), but with a main focus on the nematode communities. The meiobenthic invertebrates were exposed to two concentrations of Diltiazem and Bisoprolol, of 1.8 µg.L-1 and 1.8 mg.L-1, respectively, and one concentration of polyvinyl chloride (i.e. 20 mg.kg-1), separately and mixed. The overall meiofauna abundance was significantly reduced in all treatments, mainly that of polychaetes and amphipods. Moreover, the juveniles-gravid female ratios of the nematode communities were the lowest in the 1.8 µg.L-1 Bisoprolol treatment and for the 1.8 mg.L-1 mixture of Diltiazem and microplastics, suggesting that different dosages influence the maturity status of the examined species. The demographic results were also supported by in silico approach. The simulation of molecular interactions revealed acceptable binding affinities (up to -8.1 kcal/mol) and interactions with key residues in the germ line development protein 3 and sex-determining protein from Coenorhabditis elegans. Overall, the experimental outcome strongly indicates synergistic interactions among the beta-blockers Diltiazem and Bisoprolol and the microplastic polyvinyl chloride on marine nematode communities.

Toxicity of hydroxychloroquine, a potential treatment for COVID-19, on free-living marine nematodes.

On March 2020, hydroxychloroquine (HCQ) was recommended as a treatment for COVID-19 high risk patients. Following the massive and widespread use of HCQ worldwide, a discernible high quantity is anticipated to end-up through the sewage systems in marine coastal areas. A closed microcosm study was undertaken herein for 30 days where meiobenthic nematodes were exposed to a range of HCQ concentrations (3.162, 31.62 and 63.24 µg.ml-1). After one month of exposure in HCQ, the total abundances and Shannon-Wiener index of the assemblages decreased, whereas the individual mass and the Trophic Diversity Index increased at the highest concentrations. Overall, a numerical negative impact was observed for the epistrate feeders and non-selective deposit feeders, however, this benefited to the omnivores-carnivores, and particularly to the Oncholaimids. Such responses of the nematodes 2B and the corresponding taxa are bioindicative of current- or post-COVID-19 crisis risks in relation with the bioaccumulation of HCQ in seafood.

Nematode traits after separate and simultaneous exposure to Polycyclic Aromatic Hydrocarbons (anthracene, pyrene and benzo[a]pyrene) in closed and open microcosms.

The majority of experimental studies carried out to date, regarding the effects of pollutants on meiofauna have been conducted by means of closed systems, and rarely using open ones. The current work explored the impact of three Polycyclic Aromatic Hydrocarbons (PAHs), anthracene, pyrene and benzo[a]pyrene, applied alone or combined, on meiobenthic nematodes using both systems. The results revealed that single PAHs impacted the nematofauna similarly in closed or open systems with a higher toxicity observed for benzo[a]pyrene. However, the closed microcosms contaminated with PAHs became organically enriched, resulting in more non-selective deposit feeders and omnivores-carnivores. Taxonomic and functional effects related to combinations of PAHs were close to those of individual treatments in closed systems, however, for open ones, the outcomes were different. The caudal morphology influenced the response of taxa during their avoidance/endurance of hydrocarbons in open systems where the effects of PAHs mixtures appeared not only additive but also synergetic. Based on the results of the study, the use of open systems is preferred to closed ones as the research outcomes were more accurate and representing better conditions prevailing in nature.

First Report of Filamentous Phages Isolated from Tunisian Orchards to Control Erwinia amylovora.

Newly discovered Erwinia amylovora phages PEar1, PEar2, PEar4 and PEar6 were isolated from three different orchards in North Tunisia to study their potential as biocontrol agents. Illumina sequencing revealed that the PEar viruses carry a single-strand DNA genome between 6608 and 6801 nucleotides and belong to the Inoviridae, making them the first described filamentous phages of E. amylovora. Interestingly, phage-infected cells show a decreased swimming and swarming motility and a cocktail of the four phages can significantly reduce infection of E. amylovora in a pear bioassay, potentially making them suitable candidates for phage biocontrol.

Chemical components, antioxidant potential and hepatoprotective effects of Artemisia campestris essential oil against deltamethrin-induced genotoxicity and oxidative damage in rats.

In the present study, we evaluated the antioxidant potential of Artemisia campestris essential oil (ACEO) and the possible protective effects against deltamethrin induced hepatic toxic effects. The ACEO showed radical scavenging activity with IC50 = 47.66 ± 2.51 µg/ml, ferric reducing antioxidant power (FRAP) potential (EC50 = 5.36 ± 0.77 µg/ml), superoxide scavenging activity (IC50 = 0.175 ± 0.007 µg/ml) and ˙OH scavenging activity (IC50 = 0.034 ± 0.007 µg/ml). The obtained results of phenolic profile demonstrated that phenolic compounds are the major contributor to the antioxidant activity of ACEO. GC-MS analysis revealed the presence of 61 components in which monoterpene hydrocarbons constitute the major fraction (38.85%). In in vivo study, deltamethrin exposure caused an increase of serum AST, ALT and ALP activities, hepatic malondialdehyde (MDA) (measured as TBARS) and conjugated dienes markers of lipid peroxidation (LPO), while antioxidant enzyme activities (SOD, CAT and GPx) decreased significantly. Furthermore, it induces DNA damage as indicated by DNA fragmentation accompanied with severe histological changes in the liver tissues. The treatment with vitamin E or ACEO significantly improved the hepatic toxicity induced by deltamethrin. It can be concluded that vitamin E and ACEO are able to improve the hepatic oxidative damage induced by deltamethrin. Therefore, ACEO is an important product in reducing the toxic effects of deltamethrin.

A Bacillus subtilis strain as probiotic in poultry: selection based on in vitro functional properties and enzymatic potentialities.

We have proposed and validate an in vitro probiotic selection, based on enzymatic potentialities associated to well-established probiotic functional properties. A new Bacillus subtilis HB2 isolate, selected based on its high extracellular enzyme production, was chosen as a probiotic candidate for application as animal feed supplement. The HB2 strain showed an excellent acid and bile salts tolerance, a strong adhesion to chick enterocytes and produced antimicrobials against pathogens. An in vivo trial in poultry farming was conducted to evaluate the HB2 probiotic performance. After 35 days, HB2 achieved the higher growth performance than the control groups. The mortality and the feed conversion ratio were significantly decreased. Finally, the HB2 treated group showed wet litter and less severe ammonia odor in the atmosphere. Our study provides new insights into the importance of enzymatic potentialities, associated with the common functional properties, as a novel approach for probiotic selection.

Investigation of Antiulcer and Antioxidant Activity of Juniperus phoenicea L. (1753) Essential Oil in an Experimental Rat Model.

Juniperus phoenicea is a tree of the Cupressaceae family that is popularly known in the south of Tunisia because of its wide application in herbal medicine, including the use of its leaves to treat many diseases such as diarrhea, rheumatism, and intestinal disorders. The aim of this study was to evaluate the ulceroprotective and antioxidant activity of essential oil extracted from the leaves of J. phoenicea (EOJp) against hydrogen chloride (HCl)/ethanol-induced ulcers in rats. The antiulcer activities of 50, 75 and 100 mg/kg body weight (b.w.) EOJp were investigated on 0.3 M HCl/ethanol-induced ulcers in rats. The essential oil yield was 0.69% with 48 compounds; α-pinene was the principal component (20.24%). In vivo pretreatment with EOJp given orally provided dose-dependent protection against HCl/ethanol-induced gastric ulcers in rats. Furthermore, pretreatment with EOJp significantly decreased malondialdehyde (MDA) content and increased the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). The activity of the antiulcerogenic EOJp could be from synergistic antioxidant and anti-secretory effects. Oral use of EOJp has excellent preventive effects on induced gastric ulcers comparable to those of the proton pump inhibitor (PPI) omeprazole.

Effects of Thymus hirtus sp. algeriensis Boiss. et Reut. (Lamiaceae) essential oil on healing gastric ulcers according to sex.

BACKGROUND: Thymus algeriensis Boiss. et Reut. (Lamiaceae), popularly known as "mougecha" or "mazoukcha" is prolific in Mediterranean regions, mostly in North Africa, and is used in folk medicine to treat of stomach diseases. METHODS: In this study, animals were induced with gastric ulcers using HCl/ethanol (0.3 M HCl/60% ethanol) and treated orally with essential oil of Thymus algeriensis (EOTa) in various doses ranging from 54 mg/kg body weight to 180 mg/kg body weight. RESULT: The dose found to be effective was 180 mg/kg body weight, since this dose brought about a maximum reduction in lesion index in female rats. In gastric tissues, levels of total glutathiones (GSH, GST and GPx) and thiobarbituric acid reactive substances (TBARS) were evaluated. The activities of the antioxidant enzymes, catalase (CAT) and superoxide dismutase (SOD) were measured. Histopathological changes were observed using a cross section of gastric tissue. Chemical analysis revealed the presence of 13 components accounting for 77.7% of the essential oil from dried leaves. Oral administration of EOTa (54, 117 and 180 ml/kg) inhibited HCl/ethanol-induced ulcers. Lesion index was significantly reduced in ulcer induced animals treated with EOTa (HCl/ethanol + EOTa) compared to those ulcerated with HCl/ethanol but with no treatment given. Females showed a greater resistance to ulcers and gastric lesions occurred less often than in males. GSH, pH, enzymic antioxidants, and adherent mucus content were all significantly increased. CONCLUSION: From the data presented in this study, it can be concluded that male rats are more sensitive to gastric ulcers induced by HCl/ethanol than females.

Do trace metals (chromium, copper, and nickel) influence toxicity of diesel fuel for free-living marine nematodes?

The objective of this study was to test the hypotheses that (1) free-living marine nematodes respond in a differential way to diesel fuel if it is combined with three trace metals (chromium, copper, and nickel) used as smoke suppressants and that (2) the magnitude of toxicity of diesel fuel differs according to the level of trace metal mixture added. Nematodes from Sidi Salem beach (Tunisia) were subjected separately for 30 days to three doses of diesel fuel and three others of a trace metals mixture. Simultaneously, low-dose diesel was combined with three amounts of a trace metal mixture. Results from univariate and multivariate methods of data evaluation generally support our initial hypothesis that nematode assemblages exhibit various characteristic changes when exposed to different types of disturbances; the low dose of diesel fuel, discernibly non-toxic alone, became toxic when trace metals were added. For all types of treatments, biological disturbance caused severe specific changes in assemblage structure. For diesel fuel-treated microcosms, Marylynnia bellula and Chromaspirinia pontica were the best positive indicative species; their remarkable presence in given ecosystem may predict unsafe seafood. The powerful toxicity of the combination between diesel fuel and trace metals was expressed with only negative bioindicators, namely Trichotheristus mirabilis, Pomponema multipapillatum, Ditlevsenella murmanica, Desmodora longiseta, and Bathylaimus capacosus. Assemblages with high abundances of these species should be an index of healthy seafood. When nematodes were exposed to only trace metals, their response looks special with a distinction of a different list of indicative species; the high presence of seven species (T. mirabilis, P. multipapillatum, Leptonemella aphanothecae, D. murmanica, Viscosia cobbi, Gammanema conicauda, and Viscosia glabra) could indicate a good quality of seafood and that of another species (Oncholaimellus mediterraneus) appeared an index of the opposite situation.