Behavioural determinants of malaria risk, prevention, and care-seeking behaviours among forest-goers in Cambodia.

BACKGROUND: Cambodia has made significant progress towards achieving malaria elimination by 2025. Cases continue to decrease and are primarily concentrated in forested areas. Forest-goers are most at risk of malaria due to their proximity to the forest, poor sleeping conditions, frequent mobility, and distance from health services. Consistent use of long-lasting insecticidal nets or hammock nets (LLINs/LLIHNs), early diagnosis and treatment of cases are central to reducing disease burden. The aim of this study was to understand forest-goers' knowledge, attitudes, and practices related to malaria prevention and care-seeking, and to identify key behavioural determinants of LLIN/LLIHN use and prompt care-seeking within 24 h of developing a fever. METHODS: A mixed-methods study design consisting of a cross-sectional survey and qualitative in-depth interviews was implemented in two Cambodian provinces. Survey participants (N = 654) were recruited using respondent driven sampling. Interview participants (N = 28) were selected using purposive sampling. Findings from the survey were analysed using univariate and bivariate analysis and multivariate weighted logistic regression. Interviews were coded and analysed using thematic content analysis. RESULTS: All study participants had heard of malaria and 98% knew that malaria was transmitted by mosquitoes. LLIN/LLIHN ownership was high (94%). Although 99% of participants perceived LLIN/LLIHN use as an important malaria prevention measure, only 76% reported using one during their last visit to the forest. Only 39% of survey participants who reported seeking care did so within the recommended 24 h from fever onset during their last febrile illness. Among all study participants, 43% did not seek any healthcare during their last febrile episode. In controlled regression models, perceived community social norms were significantly associated with LLIN/LLIHN use (OR: 2.7, 96% CI 1.99-2.64) and care-seeking within 24 h of fever onset (OR: 1.7, 95% CI 1.00-2.88). Social support from other forest-goers was also significantly associated with LLIN/LLIHN use (OR: 4.9, 95% CI 1.32-18.12). CONCLUSIONS: Study findings are consistent with other studies on LLIN/LLIHN use and care-seeking behaviours. While rates of LLIN/LLIHN ownership were high among the study population, rates of use were not as high. More concerning were the delayed care-seeking behaviours. Social behaviour change activities should incorporate social norms and social support as mechanisms for behaviour change given the identified positive correlations with LLIN/LLIHN use and prompt care-seeking.

An Eco-Friendly Approach to the Control of Pathogenic Microbes and Anopheles stephensi Malarial Vector Using Magnesium Oxide Nanoparticles (Mg-NPs) Fabricated by Penicillium chrysogenum.

The discovery of eco-friendly, rapid, and cost-effective compounds to control diseases caused by microbes and insects are the main challenges. Herein, the magnesium oxide nanoparticles (MgO-NPs) are successfully fabricated by harnessing the metabolites secreted by Penicillium chrysogenum. The fabricated MgO-NPs were characterized using UV-Vis, XRD, TEM, DLS, EDX, FT-IR, and XPS analyses. Data showed the successful formation of crystallographic, spherical, well-dispersed MgO-NPs with sizes of 7-40 nm at a maximum wavelength of 250 nm. The EDX analysis confirms the presence of Mg and O ions as the main components with weight percentages of 13.62% and 7.76%, respectively. The activity of MgO-NPs as an antimicrobial agent was investigated against pathogens Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans, and exhibited zone of inhibitions of 12.0 ± 0.0, 12.7 ± 0.9, 23.3 ± 0.8, 17.7 ± 1.6, and 14.7 ± 0.6 mm respectively, at 200 µg mL-1. The activity is decreased by decreasing the MgO-NPs concentration. The biogenic MgO-NPs exhibit high efficacy against different larvae instar and pupa of Anopheles stephensi, with LC50 values of 12.5-15.5 ppm for I-IV larvae instar and 16.5 ppm for the pupa. Additionally, 5 mg/cm2 of MgO-NPs showed the highest protection percentages against adults of Anopheles stephensi, with values of 100% for 150 min and 67.6% ± 1.4% for 210 min.

Endophytic actinomycetes Streptomyces spp mediated biosynthesis of copper oxide nanoparticles as a promising tool for biotechnological applications.

In this study, two endophytic actinomycetes isolates Oc-5 and Acv-11, were isolated from healthy leaves of medicinal plant Oxalis corniculata L. These isolates were identified as Streptomyces zaomyceticus Oc-5 and Streptomyces pseudogriseolus Acv-11 using 16S rRNA gene sequence. Biomass extract of these strains were used as a greener attempt for synthesis of copper oxide nanoparticles (CuO-NPs). The synthesized NPs were characterized by UV-Vis spectroscopy, Fourier transform infra-red (FT-IR) spectroscopy, X-ray diffraction (XRD)' transmission electron microscopy (TEM), energy dispersive X-ray (EDX) and X-ray photoelectron spectroscopy (XPS). Green synthesized NPs showed surface plasmon resonance (SPR) absorption band at 400 nm, crystalline nature, spherical-shaped with an average size of 78 nm and 80.0 nm for CuO-NPs synthesized using strain Oc-5 and Acv-11, respectively. The bioactivities of CuO-NPs were evaluated. Results revealed that CuO-NPs exhibited promising antimicrobial activity against prokaryotic and eukaryotic microbial cells (Gram positive bacteria, Gram negative bacteria, unicellular and multicellular fungi). In addition, it showed antimicrobial potential against phyto-pathogenic fungal strains Fusarium oxysporum, Pythium ultimum, Aspergillus niger and Alternaria alternata. We further explored the in vitro antioxidant activity and cytotoxicity for biosynthesized CuO-NPs. The results revealed that' scavenging and total antioxidant activity for NPs synthesized using Streptomyces pseudogriseolus Acv-11 was better than those synthesized by Streptomyces zaomyceticus Oc-5. Also, the morphological changes and cell viability for Vero and Caco-2 cell line due to NPs treatments were assessed using MTT assay method. Furthermore, Larvicidal efficacy against Musca domestica and Culex pipiens was evaluated. The results obtained in this study clearly showed that biosynthesized CuO-NPs exhibited effective bioactivity and, therefore, provide a base for the development of versatile biotechnological applications soon.

Evaluating the efficacy of probiotic bacterial strain Lactobacillus plantarum for inhibition of fungal strains associated with historical manuscript deterioration: An experimental study.

The aim of this study is to develop safe biological methods for controlling fungal deterioration of historical manuscripts. Therefore, fifteen fungal isolates were obtained from paper sheets and leather skins of a deteriorated historical manuscript (dated back to the 13th century). Those isolates were identified using both traditional methods and ITS-sequencing analysis. Aspergillus niger accounted for seven strains, Penicillium citrinum for one strain, Aspergillus flavus for three, Aspergillus fumigatus for one, Aspergillus nidulans for one, and Penicillium chrysogenum for two of the fungal strains that were obtained. The ability of fungal strains for the secretion of cellulase, amylase, gelatinase, and pectinase as hydrolytic enzymes was evaluated. The capability of the probiotic-bacterial strain Lactobacillus plantarum DSM 20174 for inhibition of fungal strains that cause severe deterioration was studied using ethyl acetate-extract. The metabolic profile of the ethyl acetate-extract showed the presence of both high- and low-molecular-weight active compounds as revealed by GC-MS analysis. The safe dose to prevent fungal growth was determined by testing the ethyl acetate extract's biocompatibility against Wi38 and HFB4 as normal cell lines. The extract was found to have a concentration-dependent cytotoxic impact on Wi38 and HFB4, with IC50 values of 416 ± 4.5 and 349.7 ± 5.9 µg mL-1, respectively. It was suggested that 100 µg mL-1 as a safe concentration could be used for paper preservation. Whatman filter paper treated with ethyl acetate extract was used to cultivate the fungal strain Penicillium citrinum AX2. According to data analysis, fungal inhibition measurement, SEM, ATR-FT-IR, XRD, color change measurement, and mechanical property assessment, the recommended concentration of ethyl acetate extract was adequate to protect paper inoculated with the highest enzymatic producer fungi, P. citrinum AX2.

Exploring the Antimicrobial, Antioxidant, and Antiviral Potential of Eco-Friendly Synthesized Silver Nanoparticles Using Leaf Aqueous Extract of Portulaca oleracea L.

Herein, the prospective applications of green fabricated silver nanoparticles (Ag-NPs) within the biomedical field were investigated. The leaf aqueous extract of Portulaca oleracea L., a safe, cheap, and green method, was used to fabricate Ag-NPs. The maximum plasmon resonance of synthesized NPs has appeared at 420 nm. The various biomolecules present in the plant extract to assemble spherical Ag-NPs with sizes of 5-40 nm were analyzed using Fourier transform infrared and transmission electron microscopy. The Ag was the major content of the formed Ag-NPs with an atomic percent of 54.95% and weight percent of 65.86%, as indicated by EDX. The crystallographic structure of synthesized NPs was confirmed by the diffraction of the X-ray. The dynamic light scattering exhibits the homogeneity and mono-dispersity nature with a polydispersity index of 0.37 in the colloidal fluid and a zeta potential value of -36 mV. The synthesized Ag-NPs exhibited promising antimicrobial efficacy toward various prokaryotic and eukaryotic pathogenic microorganisms with low MIC values of 12.5 µg mL-1 and 6.25 µg mL-1, respectively. Additionally, the P. oleracea-formed Ag-NPs showed optimistic antioxidant activity assessed by DPPH and H2O2 assay methods with the highest scavenging percentages of 88.5 ± 2.3% and 76.5 ± 1.7%, respectively, at a concentration of 200 µg mL-1. Finally, the biosynthesized Ag-NPs showed high antiviral properties toward the hepatitis A virus and Cox-B4 with inhibition percentages of 79.16 ± 0.5% and 73.59 ± 0.8%, respectively. Overall, additional research is essential to explore the Ag-NP-based aqueous extract of P. oleracea for human health. In the current investigation the use of synthesized Ag-NPs as antimicrobial, antioxidant, and antiviral agents to protect against pathogenic microbes, degenerative diseases caused by various oxidative stresses, and deadly viruses is recommended.

Impact of targeted drug administration and intermittent preventive treatment for forest goers using artesunate-pyronaridine to control malaria outbreaks in Cambodia.

INTRODUCTION: The national malaria programme of Cambodia targets the rapid elimination of all human malaria by 2025. As clinical cases decline to near-elimination levels, a key strategy is the rapid identification of malaria outbreaks triggering effective action to interrupt local transmission. We report a comprehensive, multipronged management approach in response to a  2022 Plasmodium falciparum outbreak in Kravanh district, western Cambodia. METHODS: The provincial health department of Pursat in conjunction with the Center for Parasitology, Entomology and Malaria Control (CNM) identified villages where transmission was occurring using clinical records, and initiated various interventions, including the distribution of insecticide-treated bed nets, running awareness campaigns, and implementing fever screening with targeted drug administration. Health stations were set up at forest entry points, and later, targeted drug administrations with artesunate-pyronaridine (Pyramax) and intermittent preventive treatment for forest goers (IPTf) were implemented in specific village foci. Data related to adherence and adverse events from IPTf and TDA were collected. The coverage rates of interventions were calculated, and local malaria infections were monitored. RESULTS: A total of 942 individuals were screened through active fever surveillance in villages where IPTf and TDA were conducted. The study demonstrated high coverage and adherence rates in the targeted villages, with 92% (553/600) coverage in round one and 65% (387/600) in round two. Adherence rate was 99% (551/553) in round one and 98% (377/387) in round two. The study found that forest goers preferred taking Pyramax over repeated testing consistent with the coverage rates: 92% in round one compared to 65% in round two. All individuals reachable through health stations or mobile teams reported complete IPTf uptake. No severe adverse events were reported. Only six individuals reported mild adverse events, such as loss of energy, fever, abdominal pain, diarrhoea, and muscle aches. Two individuals attributed their symptoms to heavy alcohol intake following prophylaxis. CONCLUSIONS: The targeted malaria outbreak response demonstrated high acceptability, safety, and feasibility of the selected interventions. Malaria transmission was rapidly controlled using the available community resources. This experience suggests the effectiveness of the programmatic response for future outbreaks.

An Eco-Friendly Approach Utilizing Green Synthesized Titanium Dioxide Nanoparticles for Leather Conservation against a Fungal Strain, Penicillium expansum AL1, Involved in the Biodeterioration of a Historical Manuscript.

The main hypothesis of the present research is investigating the efficacy of titanium oxide nanoparticles (TiO2-NPs) to prevent the growth of fungal strains when applied on leather under an experimental study. Therefore, fifteen fungal strains were isolated from a deteriorated historical manuscript (papers and leathers) and identified by traditional methods and ITS sequence analysis, including Aspergillus chevalieri (one isolate), A. nidulans (two strains), A. flavus (four strains), A. cristatus (one strain), A. niger (one strain), Paecilomyces fulvus (two strains), Penicillium expansum (two strains), and P. citrinum (two strains). The enzymes cellulase, amylase, pectinase, and gelatinase, which play a crucial role in biodegradation, were highly active in these fungal strains. TiO2-NPs were formed using the cell-free filtrate of the probiotic bacterial strain, Lactobacillus plantarum, and characterized. Data showed that the TiO2-NPs were successfully formed with a spherical shape and anatase phase with sizes of 2-8 nm. Moreover, the EDX analysis revealed that the Ti and O ions occupied the main component with weight percentages of 41.66 and 31.76%, respectively. The in vitro cytotoxicity of TiO2-NPs toward two normal cell lines, WI38 and HFB4, showed a low toxicity effect against normal cells (IC50 = 114.1 ± 8.1µg mL-1 for Wi38, and 237.5 ± 3.5µg mL-1 for HFB4). Therefore, concentrations of 100 µg mL-1 were used to load on prepared leather samples before inoculation with fungal strain P. expansum AL1. The experimental study revealed that the loaded TiO2-NPs have the efficacy to inhibit fungal growth with percentages of 73.2 ± 2.5%, 84.2 ± 1.8%, and 88.8 ± 0.6% after 7, 14, and 21 days, respectively. Also, the analyses including SEM, FTIR-ART, color change, and mechanical properties for leather inoculated with fungal strain AL1 in the absence of NPs showed high damage aspects compared to those inoculated with fungal strains in the presence of TiO2-NPs.

Green Synthesis of Zinc Oxide Nanoparticles Using an Aqueous Extract of Punica granatum for Antimicrobial and Catalytic Activity.

The peel aqueous extract of Punica granatum was utilized to fabricate zinc oxide nanoparticles (ZnO-NPs) as a green approach. The synthesized NPs were characterized by UV-Vis spectroscopy, Fourier transform infrared (FT-IR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy, which was attached to an energy dispersive X-ray (SEM-EDX). Spherical, well arranged, and crystallographic structures of ZnO-NPs were formed with sizes of 10-45 nm. The biological activities of ZnO-NPs, including antimicrobial and catalytic activity for methylene blue dye, were assessed. Data analysis showed that the antimicrobial activity against pathogenic Gram-positive and Gram-negative bacteria, as well as unicellular fungi, was observed to occur in a dose-dependent manner, displaying varied inhibition zones and low minimum inhibitory concentration (MIC) values in the ranges of 6.25-12.5 µg mL-1. The degradation efficacy of methylene blue (MB) using ZnO-NPs is dependent on nano-catalyst concentration, contact time, and incubation condition (UV-light emission). The maximum MB degradation percentages of 93.4 ± 0.2% was attained at 20 µg mL-1 after 210 min in presence of UV-light. Data analysis showed that there is no significant difference between the degradation percentages after 210, 1440, and 1800 min. Moreover, the nano-catalyst showed high stability and efficacy to degrade MB for five cycles with decreasing values of 4%. Overall, P. granatum-based ZnO-NPs are promising tools to inhibit the growth of pathogenic microbes and degradation of MB in the presence of UV-light emission.

Antifungal Activity of Cell-Free Filtrate of Probiotic Bacteria Lactobacillus rhamnosus ATCC-7469 against Fungal Strains Isolated from a Historical Manuscript.

Herein, twelve fungal strains were isolated from a deteriorated historical manuscript dated back to the 18th century. The obtained fungal strains were identified, using the traditional method and ITS sequence analysis, as Cladosporium herbarum (two strains), Aspergillus fumigatus (five strains), A. ustus (one strain), A. flavus (two strains), A. niger (one strain), and Penicillium chrysogenum (one strain). The ability of these fungal strains to degrade the main components of the paper was investigated by their activity to secrete extracellular enzymes including cellulase, amylase, gelatinase, and pectinase. The cell-free filtrate (CFF) ability of the probiotic bacterial strain Lactobacillus rhamnosus ATCC-7469 to inhibit fungal growth was investigated. The metabolic profile of CFF was detected by GC-MS analysis, which confirmed the low and high molecular weight of various active chemical compounds. The safe dose to be used for the biocontrol of fungal growth was selected by investigating the biocompatibility of CFF and two normal cell lines, Wi38 (normal lung tissue) and HFB4 (normal human skin melanocyte). Data showed that the CFF has a cytotoxic effect against the two normal cell lines at high concentrations, with IC50 values of 525.2 ± 9.8 and 329.1 ± 4.2 µg mL-1 for Wi38 and HFB4, respectively. The antifungal activity showed that the CFF has promising activity against all fungal strains in a concentration-dependent manner. The highest antifungal activity (100%) was recorded for a concentration of 300 µg mL-1 with a zone of inhibition (ZOI) in the ranges of 21.3 ± 0.6 to 17.7 ± 0.5 mm. At a concentration of 100 µg mL-1, the activity of CFF remained effective against all fungal strains (100%), but its effectiveness decreased to only inhibit the growth of eight strains (66%) out of the total at 50 µg mL-1. In general, probiotic bacterial strains containing CFF are safe and can be considered as a potential option for inhibiting the growth of various fungal strains. It is recommended that they be used in the preservation of degraded historical papers.

Light enhanced the antimicrobial, anticancer, and catalytic activities of selenium nanoparticles fabricated by endophytic fungal strain, Penicillium crustosum EP-1.

Selenium nanoparticles (Se-NPs) has recently received great attention over owing to their superior optical properties and wide biological and biomedical applications. Herein, crystallographic and dispersed spherical Se-NPs were green synthesized using endophytic fungal strain, Penicillium crustosum EP-1. The antimicrobial, anticancer, and catalytic activities of biosynthesized Se-NPs were investigated under dark and light (using Halogen tungsten lamp, 100 Watt, λ > 420 nm, and light intensity of 2.87 W m-2) conditions. The effect of Se-NPs was dose dependent and higher activities against Gram-positive and Gram-negative bacteria as well different Candida spp. were attained in the presence of light than obtained under dark conditions. Moreover, the viabilities of two cancer cells (T47D and HepG2) were highly decreased from 95.8 ± 2.9% and 93.4 ± 3.2% in dark than those of 84.8 ± 2.9% and 46.4 ± 3.3% under light-irradiation conditions, respectively. Significant decreases in IC50 values of Se-NPs against T47D and HepG2 were obtained at 109.1 ± 3.8 and 70.4 ± 2.5 µg mL-1, respectively in dark conditions than 19.7 ± 7.2 and 4.8 ± 4.2 µg mL-1, respectively after exposure to light-irradiation. The photoluminescence activity of Se-NPs revealed methylene blue degradation efficiency of 89.1 ± 2.1% after 210 min under UV-irradiation compared to 59.7 ± 0.2% and 68.1 ± 1.03% in dark and light conditions, respectively. Moreover, superior stability and efficient MB degradation efficiency were successfully achieved for at least five cycles.

Fungal Biodeterioration of a Historical Manuscript Dating Back to the 14th Century: An Insight into Various Fungal Strains and Their Enzymatic Activities.

This study aims to assess the deterioration aspects of a historical manuscript dating back to the 14th century that was deposited in the Library of the Arabic Language Academy, Cairo, Egypt. The study aims at the exploration of the role of various fungal strains that had colonized this deteriorated manuscript in its biodeterioration through their efficacy in the secretion of various hydrolytic enzymes. To evaluate the deterioration, various techniques, including visual inspection, attenuated total reflectance Fourier transform infrared (ATR-FTIR), scanning electron microscopy (SEM), X-Ray diffraction analysis (XRD), color change, and pH value, were utilized. The fungal strains linked to the historical document were isolated, identified, and evaluated for their deterioration activities. The findings demonstrate that the manuscript exhibits a variety of deterioration signs including color change, brittleness and weakness, erosion, and removal of the grain surface pattern in leather binding. According to the ATR-FTIR, the chemical composition of the historical paper and leather underwent some alterations. The historical paper has a lower level of cellulose crystallinity than the control sample. Penicillium chrysogenum (two isolates), P. citrinum (four isolates), Aspergillus ustus (three isolates), A. terreus (two isolates), A. chinensis (one isolate), Paecilomyces sp. (one isolate), and Induratia sp. (one isolate) were among the fourteen fungal strains identified as being associated with the historical manuscript. These fungal strains produced several hydrolytic enzymes with high activity, such as cellulase, amylase, gelatinase, and pectinase, which play a key role in biodegradation.

Enhanced Antimicrobial, Cytotoxicity, Larvicidal, and Repellence Activities of Brown Algae, Cystoseira crinita-Mediated Green Synthesis of Magnesium Oxide Nanoparticles.

Herein, the metabolites secreted by brown algae, Cystoseira crinita, were used as biocatalyst for green synthesis of magnesium oxide nanoparticles (MgO-NPs). The fabricated MgO-NPs were characterized using UV-vis spectroscopy, Fourier transforms infrared spectroscopy (FT-IR), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy linked with energy-dispersive X-ray (SEM-EDX), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). Data showed successful formation of crystallographic and spherical MgO-NPs with sizes of 3-18 nm at a maximum surface plasmon resonance of 320 nm. Moreover, EDX analysis confirms the presence of Mg and O in the sample with weight percentages of 54.1% and 20.6%, respectively. Phyco-fabricated MgO-NPs showed promising activities against Gram-positive bacteria, Gram-negative bacteria, and Candida albicans with MIC values ranging between 12.5 and 50 µg mL-1. The IC50 value of MgO-NPs against cancer cell lines (Caco-2) was 113.4 µg mL-1, whereas it was 141.2 µg mL-1 for normal cell lines (Vero cell). Interestingly, the green synthesized MgO-NPs exhibited significant larvicidal and pupicidal activity against Musca domestica. At 10 µg mL-1 MgO-NPs, the highest mortality percentages were 99.0%, 95.0%, 92.2%, and 81.0% for I, II, III instars' larvae, and pupa of M. domestica, respectively, with LC50 values (3.08, 3.49, and 4.46 µg mL-1), and LC90 values (7.46, 8.89, and 10.43 µg mL-1), respectively. Also, MgO-NPs showed repellence activity for adults of M. domestica at 10 µg mL-1 with 63.0%, 77.9%, 84.9%, and 96.8% after 12, 24, 48, and 72 h, respectively.

Alleviate the Drought Stress on Triticum aestivum L. Using the Algal Extracts of Sargassum latifolium and Corallina elongate Versus the Commercial Algal Products.

Herein, two seaweed extracts (Sargassum latifolium and Corallina elongate), and two commercial seaweed products (Canada power and Oligo-X) with a concentration of 5% were used to alleviate the drought stress on wheat plants. The extract of C. elongate had the highest capacity to ameliorate the deleterious effects of water scarcity followed by S. latifolium and the commercial products. The drought stress reduced wheat shoots length and the contents of pigments (chlorophyll and carotenoids), carbohydrates, and proteins. While the highest increment in the total carbohydrates and protein contents of the wheat shoot after two stages, 37-and 67-days-old, were noted in drought-stressed plants treated with C. elongate extract with values of (34.6% and 22.8%) and (51.9% and 39.5%), respectively, compared to unstressed plants. Decreasing the activity of antioxidant enzymes, peroxidase, superoxidase dismutase, and polyphenol oxidase in drought-stressed plants treated with algal extracts indicated amelioration of the response actions. Analysis of phytohormones in wheat plants exhibited increasing GA3 and IAA contents with percentages of (20.3-13.8%) and (72.7-25%), respectively. Interestingly, all morphological and metabolic characteristics of yield were improved due to the algal treatments compared with untreated drought-stressed plants. Overall, the algal extracts, especially those from seaweed of C. elongate, could represent a sustainable candidate to overcome the damage effects of water deficiency in the wheat plant.

Molecular biodiversity of arbuscular mycorrhizal fungi in trace metal-polluted soils.

We assessed the indigenous arbuscular mycorrhizal fungi (AMF) community structure from the roots and associated soil of Plantago major (plantain) plants growing on sites polluted with trace metals (TM) and on unpolluted sites. Uncontaminated and TM-contaminated sites containing As, Cd, Cu, Pb, Sn and Zn were selected based on a survey of the TM concentration in soils of community gardens in the City of Montréal. Total genomic DNA was extracted directly from these samples. PCR followed by denaturing gradient gel electrophoresis (PCR-DGGE), augmented by cloning and sequencing, as well as direct sequencing techniques, was all used to investigate AMF community structure. We found a decreased diversity of native AMF (assessed by the number of AMF ribotypes) in soils and plant roots harvested from TM-polluted soils compared with unpolluted soils. We also found that community structure was modified by TM contamination. Various species of Glomus, Scutellospora aurigloba and S. calospora were the most abundant ribotypes detected in unpolluted soil; ribotypes of G. etunicatum, G. irregulare/G. intraradices and G. viscosum were found in both polluted and unpolluted soils, while ribotypes of G. mosseae and Glomus spp. (B9 and B13) were dominant in TM-polluted soils. The predominance of G. mosseae in metal-polluted sites suggests the tolerance of this species to TM stress, as well as its potential use for phytoremediation. These data are relevant for our understanding of how AMF microbial communities respond to natural environments that contain a broad variety of toxic inorganic compounds and will substantially expand our knowledge of AMF ecology and biodiversity.

Plant growth-promoting properties of bacterial endophytes isolated from roots of Thymus vulgaris L. and investigate their role as biofertilizers to enhance the essential oil contents.

The main objective of the current study was to improve the essential oil contents of Thymus vulgaris L. using bio-inoculation with bacterial endophytes. Therefore, out of fourteen endophytic bacterial isolates obtained from roots of T. vulgaris, five isolates were selected based on the highest nitrogen-fixation and phosphate solubilization activity and identified as: Bacillus haynesii T9r, Citrobacter farmeri T10r, Bacillus licheniformis T11r, Bacillus velezensis T12r, and Bacillus velezensis T13r. These five strains have been recorded as ammonia, hydrogen cyanide (HCN), siderophores, and indole-3-acetic acid (IAA) producers. These strains have the efficacy to fix-nitrogen by reduction of acetylene with values of 82.133±1.4-346.6±1.4 n-mole-C2H4/ml/24 h. The IAA, gibberellic acid, abscisic acid, benzyl, kinten, and ziaten production were confirmed using HPLC. Two strains of T11r and T13r showed the highest plant growth-promoting properties and were selected for bio-inoculation of T. vulgaris individually or in a consortium with different mineral fertilization doses (0, 50, 75, and 100%) under field conditions. The highest growth performance was attained with the endophytic consortium (T11r+T13r) in the presence of 100% mineral fertilization. The GC-MS analysis of thyme oil contents showed the presence of 23 various compounds with varying percentages and the thymol fraction represented the highest percentages (39.1%) in the presence of the bacterial consortium.

Implication of plant growth-promoting rhizobacteria of Bacillus spp. as biocontrol agents against wilt disease caused by Fusarium oxysporum Schlecht. in Vicia faba L.

Out of seven Fusarium spp. isolated from infected faba bean roots, two Fusarium oxysporum were selected and showed faba bean-wilt disease severity with percentages of 68% and 47% under greenhouse conditions. The F. oxysporum showed the highest wilt disease was selected to complete the current study. Three rhizobacterial strains were isolated and identified as Bacillus velezensis Vb1, B. paramycoides Vb3, and B. paramycoides Vb6. These strains showed the highest in-vitro antagonistic activity by the dual-culture method against selected F. oxysporum with inhibition percentages of 59±0.2, 46±0.3, and 52±0.3% for Vb1, Vb3, and Vb6, respectively. These rhizobacterial strains exhibit varied activity for nitrogen-fixing and phosphate-solubilizing. Moreover, these strains showed positive results for ammonia, HCN, and siderophores production. The phytohormones production (indole-3-acetic acid, ABA, benzyl, kinten, ziaten, and GA3) and secretion of various lytic enzymes were recorded by these strains with varying degrees. Under greenhouse conditions, the rhizobacterial strains Vb1, Vb3, Vb6, and their consortium can protect faba bean from wilt caused by F. oxysporum with percentages of 70, 60, 65, and 82%, respectively. Under field conditions, the inoculation with the rhizobacterial consortium (Vb1+Vb3+Vb6) significantly increases the growth performance of the F. oxysporum-infected faba bean plant and recorded the highest wilt protection (83.3%).

Evaluate the Toxicity of Pyrethroid Insecticide Cypermethrin before and after Biodegradation by Lysinibacillus cresolivuorans Strain HIS7.

Herein, bacterial isolate HIS7 was obtained from contaminated soil and exhibited high efficacy to degrade pyrethroid insecticide cypermethrin. The HIS7 isolate was identified as Lysinibacillus cresolivuorans based on its morphology and physiology characteristics as well as sequencing of 16S rRNA. The biodegradation percentages of 2500 ppm cypermethrin increased from 57.7% to 86.9% after optimizing the environmental factors at incubation condition (static), incubation period (8-days), temperature (35 °C), pH (7), inoculum volume (3%), and the addition of extra-carbon (glucose) and nitrogen source (NH4Cl2). In soil, L. cresolivuorans HIS7 exhibited a high potential to degrade cypermethrin, where the degradation percentage increased from 54.7 to 93.1% after 7 to 42 days, respectively. The qualitative analysis showed that the bacterial degradation of cypermethrin in the soil was time-dependent. The High-Performance Liquid Chromatography (HPLC) analysis of the soil extract showed one peak for control at retention time (R.T.) of 3.460 min and appeared three peaks after bacterial degradation at retention time (R.T.) of 2.510, 2.878, and 3.230 min. The Gas chromatography-mass spectrometry (GC-MS) analysis confirmed the successful degradation of cypermethrin by L. cresolivuorans in the soil. The toxicity of biodegraded products was assessed on the growth performance of Zea mays using seed germination and greenhouse experiment and in vitro cytotoxic effect against normal Vero cells. Data showed the toxicity of biodegraded products was noticeably decreased as compared with that of cypermethrin before degradation.

Plant Growth-Promoting Endophytic Bacterial Community Inhabiting the Leaves of Pulicaria incisa (Lam.) DC Inherent to Arid Regions.

In this study, 15 bacterial endophytes linked with the leaves of the native medicinal plant Pulicaria incisa were isolated and identified as Agrobacterium fabrum, Acinetobacter radioresistant, Brevibacillus brevis, Bacillus cereus, Bacillus subtilis, Paenibacillus barengoltzii, and Burkholderia cepacia. These isolates exhibited variant tolerances to salt stress and showed high efficacy in indole-3-acetic acid (IAA) production in the absence/presence of tryptophan. The maximum productivity of IAA was recorded for B. cereus BI-8 and B. subtilis BI-10 with values of 117 ± 6 and 108 ± 4.6 µg mL-1, respectively, in the presence of 5 mg mL-1 tryptophan after 10 days. These two isolates had a high potential in phosphate solubilization and ammonia production, and they showed enzymatic activities for amylase, protease, xylanase, cellulase, chitinase, and catalase. In vitro antagonistic investigation showed their high efficacy against the three phytopathogens Fusarium oxysporum, Alternaria alternata, and Pythium ultimum, with inhibition percentages ranging from 20% ± 0.2% to 52.6% ± 0.2% (p ≤ 0.05). Therefore, these two endophytic bacteria were used as bio-inoculants for maize seeds, and the results showed that bacterial inoculations significantly increased the root length as well as the fresh and dry weights of the roots compared to the control plants. The Zea mays plant inoculated with the two endophytic strains BI-8 and BI-10 significantly improved (p ≤ 0.05) the growth performance as well as the nutrient uptake compared with an un-inoculated plant.

Comparative Study between Exogenously Applied Plant Growth Hormones versus Metabolites of Microbial Endophytes as Plant Growth-Promoting for Phaseolus vulgaris L.

Microbial endophytes organize symbiotic relationships with the host plant, and their excretions contain diverse plant beneficial matter such as phytohormones and bioactive compounds. In the present investigation, six bacterial and four fungal strains were isolated from the common bean (Phaseolus vulgaris L.) root plant, identified using molecular techniques, and their growth-promoting properties were reviewed. All microbial isolates showed varying activities to produce indole-3-acetic acid (IAA) and different hydrolytic enzymes such as amylase, cellulase, protease, pectinase, and xylanase. Six bacterial endophytic isolates displayed phosphate-solubilizing capacity and ammonia production. We conducted a field experiment to evaluate the promotion activity of the metabolites of the most potent endophytic bacterial (Bacillus thuringiensis PB2 and Brevibacillus agri PB5) and fungal (Alternaria sorghi PF2 and, Penicillium commune PF3) strains in comparison to two exogenously applied hormone, IAA, and benzyl adenine (BA), on the growth and biochemical characteristics of the P. vulgaris L. Interestingly, our investigations showed that bacterial and fungal endophytic metabolites surpassed the exogenously applied hormones in increasing the plant biomass, photosynthetic pigments, carbohydrate and protein contents, antioxidant enzyme activity, endogenous hormones and yield traits. Our findings illustrate that the endophyte Brevibacillus agri (PB5) provides high potential as a stimulator for the growth and productivity of common bean plants.

Harnessing Bacterial Endophytes for Promotion of Plant Growth and Biotechnological Applications: An Overview.

Endophytic bacteria colonize plants and live inside them for part of or throughout their life without causing any harm or disease to their hosts. The symbiotic relationship improves the physiology, fitness, and metabolite profile of the plants, while the plants provide food and shelter for the bacteria. The bacteria-induced alterations of the plants offer many possibilities for biotechnological, medicinal, and agricultural applications. The endophytes promote plant growth and fitness through the production of phytohormones or biofertilizers, or by alleviating abiotic and biotic stress tolerance. Strengthening of the plant immune system and suppression of disease are associated with the production of novel antibiotics, secondary metabolites, siderophores, and fertilizers such as nitrogenous or other industrially interesting chemical compounds. Endophytic bacteria can be used for phytoremediation of environmental pollutants or the control of fungal diseases by the production of lytic enzymes such as chitinases and cellulases, and their huge host range allows a broad spectrum of applications to agriculturally and pharmaceutically interesting plant species. More recently, endophytic bacteria have also been used to produce nanoparticles for medical and industrial applications. This review highlights the biotechnological possibilities for bacterial endophyte applications and proposes future goals for their application.