Search results for: fungal growth inhibition

Authors: Salama A. Ouf, Amera A. El-Adly, Abdelaleam H. Mohamed

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Dermatophytosis is the infection of keratinized tissues such as hair, nail and the stratum corneum of the skin by dermatophytic fungi. Infection is generally cutaneous and restricted to the non-living cornified layers because of the inability of the fungi to penetrate the deeper tissues or organs of immunocompetent hosts. In Saudi Arabia, Onychomycosis is the most frequent infection (40.3%), followed by tinea capitis (21.9%), tinea pedis (16%), tinea cruris (15.1%), and tinea corporis (6.7%). Several azole compounds have been tried to control dermatophytic infection, however, the azole-containing medicines may interfere with the activity of hepatic microsomal enzymes, sex and thyroid hormones, and testosterone biosynthesis. In this research, antibody-conjugated nanoparticles stimulated by cold plasma and laser were evaluated in vitro against some dermatophytes isolated from the common types of tinea. Different types of nanomaterials were tested but silver nanoparticles (AgNPs) were proved to be most effective against the dermatophytes under test. The use of cold plasma coupled with antibody-conjugated nano-particles has severe impact on dermatophytes where the inhibition of growth, spore germination keratinase activity was more than 88% in the case of Trichophyton rubrum, T. violaceum, Microsprum canis and M. gypseum. Complete inhibition of growth for all dermatophytes was brought about by the interaction of conjugated nanoparticles, with cold plasma and laser treatment. The in vivo test with inoculated guinea pigs achieved promising results where the recovery from the infection reached 95% in the case of M. canis –inoculated pigs treated with AgNPs pretreated with cold plasma and laser.

Keywords: cold plasma, dermatophytes, laser, silver nanoparticles

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Authors: Parviz Almasi

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Ethylene is produced as a gaseous growth regulator in all plants and their constructive parts such as roots, stems, leaves, flowers and fruits. It is considered a multifunctional phytohormone that regulates both growths including flowering, fruit ripening, inhibition of root growth, and senescence such as senescence of leaves and flowers and etc. In addition, exposure to external ethylene is caused some changes that are often undesirable and harmful. Some flowers are more sensitive to others and when exposed to ethylene; their aging process is hastened. 1-MCP is an exogenous and endogenous ethylene action inhibitor, which binds to the ethylene receptors in the plants and prevents ethylene-dependent reactions. The binding affinity of 1- MCP for the receptors is about 10 times more than ethylene. Hence, 1-MCP can be a potential candidate for controlling of ethylene injury in horticultural crops. This review integrates knowledge of ethylene biosynthesis in the plants and also a mode of action of 1-MCP in preventing of ethylene injury.

Keywords: ethylene injury, biosynthesis, ethylene sensitivity, 1-MCP

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Authors: Veenu Bala, Yashpal S. Chhonker, Bhavana Kushwaha, Rabi S. Bhatta, Gopal Gupta, Vishnu L. Sharma

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According to UNAIDS 2013 estimate nearly 52% of all individuals living with HIV are now women of reproductive age (15–44 years). Seventy-five percent cases of HIV acquisition are through heterosexual contacts and sexually transmitted infections (STIs), attributable to unsafe sexual behaviour. Each year, an estimated 500 million people acquire atleast one of four STIs: chlamydia, gonorrhoea, syphilis and trichomoniasis. Trichomonas vaginalis (TV) is exclusively sexually transmitted in adults, accounting for 30% of STI cases and associated with pelvic inflammatory disease (PID), vaginitis and pregnancy complications in women. TV infection resulted in impaired vaginal milieu, eventually favoring HIV transmission. In the absence of an effective prophylactic HIV vaccine, prevention of new infections has become a priority. It was thought worthwhile to integrate HIV prevention and reproductive health services including unintended pregnancy protection for women as both are related with unprotected sex. Initially, nonoxynol-9 (N-9) had been proposed as a spermicidal agent with microbicidal activity but on the contrary it increased HIV susceptibility due to surfactant action. Thus, to accomplish an urgent need of novel woman controlled non-detergent microbicidal spermicides benzenepropanamine analogues have been synthesized. At first, five benzenepropanamine-dithiocarbamate hybrids have been synthesized and evaluated for their spermicidal, anti-Trichomonas and anti-fungal activities along with safety profiling to cervicovaginal cells. In order to further enhance the scope of above study benzenepropanamine was hybridized with thiourea as to introduce anti-HIV potential. The synthesized hybrid molecules were evaluated for their reverse transcriptase (RT) inhibition, spermicidal, anti-Trichomonas and antimicrobial activities as well as their safety against vaginal flora and cervical cells. simulated vaginal fluid (SVF) stability and pharmacokinetics of most potent compound versus N-9 was examined in female Newzealand (NZ) rabbits to observe its absorption into systemic circulation and subsequent exposure in blood plasma through vaginal wall. The study resulted in the most promising compound N-butyl-4-(3-oxo-3-phenylpropyl) piperazin-1-carbothioamide (29) exhibiting better activity profile than N-9 as it showed RT inhibition (72.30 %), anti-Trichomonas (MIC, 46.72 µM against MTZ susceptible and MIC, 187.68 µM against resistant strain), spermicidal (MEC, 0.01%) and antifungal activity (MIC, 3.12–50 µg/mL) against four fungal strains. The high safety against vaginal epithelium (HeLa cells) and compatibility with vaginal flora (lactobacillus), SVF stability and least vaginal absorption supported its suitability for topical vaginal application. Docking study was performed to gain an insight into the binding mode and interactions of the most promising compound, N-butyl-4-(3-oxo-3-phenylpropyl) piperazin-1-carbothioamide (29) with HIV-1 Reverse Transcriptase. The docking study has revealed that compound (29) interacted with HIV-1 RT similar to standard drug Nevirapine. It may be concluded that hybridization of benzenepropanamine and thiourea moiety resulted into novel lead with multiple activities including RT inhibition. A further lead optimization may result into effective vaginal microbicides having spermicidal, anti-Trichomonas, antifungal and anti-HIV potential altogether with enhanced safety to cervico-vaginal cells in comparison to Nonoxynol-9.

Keywords: microbicidal, nonoxynol-9, reverse transcriptase, spermicide

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Authors: Guru Prasad Satsangi, Shiv Shankar Gautam

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Allelopathy is a potential area of research for sustainable agriculture. It is environmentally safe, can conserve the available resources, and also may mitigate the problems raised by synthetic chemicals. The allelo-chemicals are secondary metabolites produced by plants, which are the byproducts of the primary metabolic process. These allelo-chemicals may be stimulatory, inhibitory, or may have no effect on the growth of the other plants. It has been observed in the present study that foliar extracts of Leucaena leucocephala showed an inhibitory effect on the germination of the test crop maize. The results revealed that at different concentrations of Leucaena leucocephala foliar extract, caused a significant inhibition in germination and growth behavior of Zea mays L. seedlings. Minimum germination and growth occurred in 100 % concentration, and an increase in extract concentrations result in a decrease in the germination. Bioassay also depicted that this inhibitory effect was proportional to the concentration of the extract as the higher concentration having a lesser stimulatory effect or vice versa. The phytochemical analysis of the secondary metabolites from foliar extracts of Leucaena leucocephala L. showed the presence of tannins, saponins, phenols, alkaloids, and flavanoids. Among various extracts, the presence of methanol extract was found in a significant amount of phytochemicals, followed by the aqueous and ethanol extracts. Leaves showed a significantly higher amount of the allelochemicals.

Keywords: allelopathic effect, germination /growth behavior , foliar extracts, Leucaena leucceophala , Zea mays L.

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Authors: Hamdi Belfeki, Belgacem Chandoul, Mnasser Hassouna, Mondher Mejri

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Aqueous and ethanolic extracts of eight Tunisian aromatic and medicinal plants (TAMP) were characterized by studying their composition in polyphenols and also their antiradical and antioxidant capacities. In absence and in the presence of the various extracts, α-amylase from Bacillus subtlis activity, was measured in order to detect a potential inhibition. The total contents of polyphenols and flavonoid vary in function of TAMP and the mobile phase used for the extraction (distilled water or ethanol). The ethanolic extracts showed the most significant antiradical and antioxidant activities. Only the extracts from Coriandrum sativum showed a significant inhibiting effect on the α-amylase activity. This inhibiting capacity could be correlated with the chemical profile of the two extracts, due to the fact that they have the greatest amount of total flavonoid. The ethanolic extract has the most important antioxidant and anti-radicalizing activities among the sixteen extracts studied. The inhibition kinetics of the two coriander extracts were evaluated by pre-incubation method, using Lineweaver-Burk’s equation, obtained by linearization of Michaeilis-Menten’s expression. The results showed that both extracts exercised a competitive inhibition mechanism.

Keywords: α-amylase, antioxidant activity, aromatic and medicinal plants, inhibition

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Authors: Himanshu Lal, Bipasha Ghosh, Arun Srivastava

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A pilot study of indoor air quality in terms of bioaerosol (fungus and bacteria) and few selective microbial volatile organic compounds (MVOCs) was carried out in different indoor sections of a library for two seasons, namely monsoon and post monsoon. Bioaerosol sampling was carried out using Anderson six stage viable sampler at a flow rate of 28.3 L/min while MVOCs were collected on activated charcoal tubes ORBOTM 90 Carboxen 564.Collected MVOCs were desorbed using carbon disulphide (CS2) and analysed by GC-FID. Microscopic identification for fungus was only carried out. Surface dust was collected by sterilised buds and cultured to identify fungal contaminants. Unlike bacterial size distribution, fungal bioaerosol concentration was found to be highest in the fourth stage in different sections of the library. In post monsoon season both fungal bioaerosol (710 to 3292cfu/m3) and bacterial bioaerosol (298 to 1475cfu/m3) were fund at much greater concentration than in monsoon. In monsoon season unlike post monsoon, I/O ratio for both the bioaerosol fractions was more than one. Rain washout could be the reason of lower outdoor concentration in monsoon season. On the contrary most of the MVOCs namely 1-hexene, 1-pentanol and 1-octen-3-ol were found in the monsoon season instead of post monsoon season with the highest being 1-hexene with 7.09µg/m3 concentration. Among the six identified fungal bioaerosol Aspergillus, Cladosporium and Penicillium were found in maximum concentration while Aspergillus niger, Curvuleria lunata, Cladosporium cladosporioides and Penicillium sp., was indentified in surface dust samples. According to regression analysis apart from environmental factors other factors also played an important role. Thus apart from outdoor infiltration and human sources, accumulated surface dust mostly on organic materials like books, wooden furniture and racks can be attributed to being one of the major sources of both fungal bioaerosols as well as MVOCs found in the library.

Keywords: bacteria, Fungi, indoor air, MVOCs

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Authors: Roland Tolulope Loto, Cleophas Akintoye Loto, Abimbola Patricia Popoola

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The inhibiting action of 2-dimethylamine on the electrochemical behaviour of austenitic stainless steel (type 304) in dilute hydrochloric was evaluated through weight-loss method, open circuit potential measurement and potentiodynamic polarization tests at specific concentrations of the organic compound. Results obtained reveal that the compound performed effectively giving a maximum inhibition efficiency of 79% at 12.5% concentration from weight loss analysis and 80.9% at 12.5% concentration from polarization tests. The average corrosion potential of -321 mV was obtained the same concentration from other tests which is well within passivation potentials on the steel thus, providing good protection against corrosion in the acid solutions. 2-dimethylamine acted through physiochemical interaction at the steel/solution interface from thermodynamic calculations and obeyed the Langmuir adsorption isotherm. The values of the inhibition efficiency determined from the three methods are in reasonably good agreement. Polarization studies showed that the compounds behaved as cathodic type inhibitor.

Keywords: corrosion, 2-dimethylamine, inhibition, adsorption, hydrochloric acid, steel

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Authors: German Galvez-Garcia, Javier Albayay, Javiera Peña, Marta Lavin, George A. Michael

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The aim of this research was to investigate whether the selection of the actions needs different inhibition processes during the response selection stage. In Experiment 1, we compared the magnitude of the Spatial Stroop effect, which occurs in response selection stage, in two motor actions (lifting vs reaching) when the participants performed both actions in the same block or in different blocks (mixed block vs. pure blocks).Within pure blocks, we obtained faster latencies when lifting actions were performed, but no differences in the magnitude of the Spatial Stroop effect were observed. Within mixed block, we obtained faster latencies as well as bigger-magnitude for Spatial Stroop effect when reaching actions were performed. We concluded that when no action selection is required (the pure blocks condition), inhibition works as a unitary system, whereas in the mixed block condition, where action selection is required, different inhibitory processes take place within a common processing stage. In Experiment 2, we investigated this common processing stage in depth by limiting participants’ available resources, requiring them to engage in a concurrent auditory task within a mixed block condition. The Spatial Stroop effect interacted with Movement as it did in Experiment 1, but it did not significantly interact with available resources (Auditory task x Spatial Stroop effect x Movement interaction). Thus, we concluded that available resources are distributed equally to both inhibition processes; this reinforces the likelihood of there being a common processing stage in which the different inhibitory processes take place.

Keywords: inhibition process, motor processes, selective inhibition, dual task

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Authors: Sobhy A. El-Sohaimy, Mohamed G. Shehata, Ashwani Mathur, Amira G. Darwish, Nourhan M. Abd El-Aziz, Pammi Gauba, Pooja Upadhyay

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Sea buckthorn is a temperate bush plant native to Asian and European countries, explored across the world in traditional medicine to treat various diseases due to the presence of an exceptionally high content of phenolics, flavonoids and antioxidants. In addition to the evaluation of nutrients and active compounds, the focus of the present work was to assess the optimal levels for L. plantarum RM1 growth by applying response surface methodology (RSM), and to determine the impact of juice fermentation on antioxidant, anti-hypertension and anticancer activity, as well as on organoleptic properties. Sea buckthorn berries were shown to contain good fiber content (6.55%, 25 DV%), high quality of protein (3.12%, 6.24 DV%) containing: histidine, valine, threonine, leucine and lysine (with AAS 24.32, 23.66, 23.09, 23.05 and 21.71%, respectively), and 4.45% sugar that pro- vides only 79 calories. Potassium was shown to be the abundant mineral content (793.43%, 22.66 DV), followed by copper and phosphorus (21.81 and 11.07 DV%, respectively). Sea buckthorn juice exhibited a rich phenolic, flavonoid and carotenoid content (283.58, 118.42 and 6.5 mg/g, respec- tively), in addition to a high content of vitamin C (322.33 mg/g). The HPLC profile indicated that benzoic acid is the dominant phenolic compound in sea buckthorn berries (3825.90 mg/kg). Antiox- idant potentials (DPPH and ABTS) of sea buckthorn showed higher inhibition than ascorbic acid. Antimicrobial potentials were most pronounced against Escherichia coli BA12296 (17.46 mm). The probiotic growth was 8.5 log cfu/mL, with juice concentration, inoculum size and temperature as the main contributors to probiotic growth with a 95% confidence level. Fermentation of sea buck- thorn juice with L. plantarum RM1 enhanced the functional phenolic and flavonoid content, as well as antioxidant and antimicrobial activities. The fermentation with L. plantarum RM1 enhanced the anti-hypertension and anticancer properties of the sea buckthorn juice and gained consumers’ sensorial overall acceptance.

Keywords: sea buckthorn juice, L. plantarum RM1, fermentation, antioxidant, antimicrobial, angiotensin converting enzyme inhibition

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Authors: S. F. Afzali, W. L. Wong

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The epizootic ulcerative syndrome (EUS) causes by the oomycete fungus, Aphanomyces invadans; known to be one of the infectious fish diseases for farmed and wild fishes in fresh and brackish-water from the Asia-pacific region, America and Africa. Although, EUS had been documented by the Office International des Epizooties (OIE) since 1995, hitherto, there is neither standard chemical agents that can be used for successful treatment of this destructive infection in the time of outbreak; nor available vaccine for prevention. Plant-based remedies in controlling fish diseases are gaining much attention recently as an alternative to chemical treatments, which possess negative effects to the environment and human. In present study, Sonneratia alba, a mangrove plant belongs to the Sonneratiaceae family, was screened in vitro and in vivo for its antifungal activity against A. invadans mycelium growth and its effects on fish innate immune system and disease resistant. The in vitro tests was performed using the disc diffusion methods with measurements of minimum inhibitory concentration (MIC) and inhibition zone. For in vivo study, the S. alba extract supplemented diets were administrated at 0.0, 1.0%, 3.0%, and 5.0% on healthy goldfish, Carassius auratus, which challenged with A. invadans zoospores (100 spores/ml). To compare the significant differences in the hematological and immunological parameters obtained from the experiments, the data were analysed using the SPSS. The methanol extract of S. alba effectively inhibited the mycelial growth of A. invadans at a minimum concentration of 1000 ppm for agar and filter paper diffusion experiments. In the agar diffusion test, 500 ppm of the extract inhibited the fungus mycelial growth up to 96 hours after exposure. The mycelial growth from the edge of the pre-inoculated A. invadans agar discs treated with S. alba extracts at concentrations of 100, 500 and 1000 ppm were 15, 8 and 0 mm respectively. The results of the filter paper disc test showed that the S. alba extract at its minimal inhibitory concentration (1000 ppm) has similar qualitative inhibitory effect as malachite green at 1 ppm and formalin at 250 ppm. According to the in vivo tests findings, in the infected fish fed with 3.0% and 5.0% supplementation diet, the numbers of white blood cell and myeloperoxidase activity significantly increased after the second week of treatment. Whilst the numbers of red blood cell significantly decreased in the infected fish fed with 0.0 and 1.0% supplementation diet. After the third week of feeding, significant increases in the total protein, albumin level, lysozyme activity were recorded in the infected fish fed with 3.0% and 5.0% supplementation diet. Also, the enriched diets increased the survival rate as compared to the untreated group that suffered from 90% mortality. The present study indicated that S. alba extract may inhibit the mycelial growth of A. invadans effectively, suggesting an alternative to other chemotherapeutic agents, which brought much environmental and health concerns to the public, for EUS treatment.

Keywords: fungal pathogen, goldfish, organic extract, treatment

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Authors: Abdulaziz Bashir Kutawa, Khairulmazmi Ahmad, Mohd Zobir Hussein, Asgar Ali, Mohd Aswad Abdul Wahab, Amara Rafi, Mahesh Tiran Gunasena, Muhammad Ziaur Rahman, Md. Imam Hossain, Syazwan Afif Mohd Zobir

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Rice is a cereal crop and belongs to the family Poaceae, it was domesticated in southern China and North-Eastern India around 8000 years ago, and it’s the staple nourishment for over half of the total world’s population. Rice production worldwide is affected by different abiotic and biotic stresses. Diseases are important challenges for the production of rice, among all the diseases in rice plants, the most severe and common disease is the rice blast. Worldwide, it is one of the most damaging diseases affecting rice cultivation, the disease is caused by the non-obligate filamentous ascomycete fungus called Magnaporthe grisae or Pyricularia oryzae Cav. Nanotechnology is a new idea to improve agriculture by combating the diseases of plants, as nanoparticles were found to possess an inhibitory effect on different species of fungi. This work aimed to develop and determine the efficacy of agronanofungicides, and commercial fungicides (in-vitro and in-vivo). The agronanofungicides were developed using ionic gelation methods. In-vitro antifungal activity of the synthesized agronanofungicides was evaluated against P. oryzae using the poisoned medium technique. The potato dextrose agar (PDA) was amended in several concentrations; 0.001, 0.005, 0.01, 0.025, 0.05, 0.1, 0.15, 0.20, 0.25, 0.30, and 0.35 ppm for the agronanofungicides. Medium with the only solvent served as a control. Mycelial growth was recorded every day, and the percentage inhibition of radial growth (PIRG) was also calculated. Based on the results of the zone of inhibition, the chitosan-hexaconazole agronanofungicide (2g/mL) was the most effective fungicide to inhibit the growth of the fungus with 100% inhibition at 0.2, 0.25, 0.30, and 0.35 ppm, respectively. The least were found to be propiconazole and basamid fungicides with 100% inhibition only at 100 ppm. In terms of the glasshouse results, the chitosan-hexaconazole-dazomet agronanofungicide (CHDEN) treatment (2.5g/L) was found to be the most effective fungicide to reduce the intensity of the disease with a disease severity index (DSI) of 19.80%, protection index (PI) of 82.26%, lesion length of 1.63cm, disease reduction (DR) of 80.20%, and AUDPC (390.60 Unit2). The least effective fungicide was found to be ANV with a disease severity index (45.60%), protection index (45.24%), lesion length (3.83 cm), disease reduction (54.40%), and AUDPC (1205.75 Unit2). The negative control did not show any symptoms during the glasshouse assay, while the untreated control treatment exhibited severe symptoms of the disease with a DSI value of 64.38%, lesion length of 5.20 cm, and AUDPC value of 2201.85 Unit2, respectively. The treatments of agronanofungicides have enhanced the yield significantly with CHDEN having 239.00 while the healthy control had 113.67 for the number of grains per panicle. The use of CHEN and CHDEN will help immensely in reducing the severity of rice blast in the fields, and this will increase the yield and profit of the farmers that produced rice.

Keywords: chitosan, dazomet, disease severity, efficacy, and blast disease

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Authors: Akwu A. Nneka, Naidoo, Yougasphree

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A significantly high number of microbes exist in higher plants; these microbes include bacteria, fungi, and actinomycetes. There are reports on the benefits of endophytic fungi and their products of metabolism to the host plant and man, consequently, it is expedient to explore the changes that could arise as a result of manipulating their growth media. Grewia lasiocarpa E. Mey. ex Harv. (Malvaceae) is an indigenous Southern African plant, that belongs to a genus with known medicinal properties. Three media were used to culture the endophytic fungi viz., Potato Dextrose Agar (PDA), Malt Extract Agar (MEA), and Bacteriological Agar (BA) were used singly, and supplemented with three dilutions of the leaves and stem bark extracts. The manipulated growth media composition had a significant effect on the diversity of the isolated fungal populations. Several endophytic fungi were isolated; their distribution and diversity revealed a significant relatedness with the manipulated media. The media supplemented with the plant extracts was observed to give a significant increase in the growth rate and yield of the endophytes. To the best of our knowledge, this is the first study describing the endophytic fungi present in the leaves and stem bark of G. lasiocarpa E. Mey. ex Harv.

Keywords: Grewia lasiocarpa, plant-based extracts, endophytic fungi, Malvaceae

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Authors: H. Elmsellem, A. Aouniti, S. Radi, A. Chetouani, B. Hammouti

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The synthesis of new organic molecules offers various molecular structures containing heteroatoms and substituents for corrosion protection in acid pickling of metals. The most synthesized compounds are the nitrogen heterocyclic compounds, which are known to be excellent complex or chelate forming substances with metals. The choice of the inhibitor is based on two considerations: first it could be synthesized conveniently from relatively cheap raw materials, secondly, it contains the electron cloud on the aromatic ring or, the electro negative atoms such as nitrogen and oxygen in the relatively long chain compounds. In the present study, (NE)‐2‐methyl‐N‐(thiophen‐2‐ylmethylidene) aniline(T) was synthesized and its inhibiting action on the corrosion of mild steel in 1 M hydrochloric acid was examined by different corrosion methods, such as weight loss, potentiodynamic polarization and electrochemical impedance spectroscopy (EIS). The experimental results suggest that this compound is an efficient corrosion inhibitor and the inhibition efficiency increases with the increase in inhibitor concentration. Adsorption of this compound on mild steel surface obeys Langmuir’s isotherm. Correlation between quantum chemical calculations and inhibition efficiency of the investigated compound is discussed using the Density Functional Theory method (DFT).

Keywords: mild steel, Schiff base, inhibition, corrosion, HCl, quantum chemical

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Authors: Syed Asif Hassan, Ritu Barthwal

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Topoisomerase I and II α plays a crucial role in the DNA-maintenance in all living cells, and for this reason, inhibitors of this enzyme have been much studied. In this paper, we have described the inhibitory effect of the aqueous extract of Picrorrhiza kurroa on human topoisomerases by measuring the relaxation of superhelical plasmid pBR322 DNA. The aqueous extract inhibited topoisomerase I and II α in a concentration-dependent manner (Inhibitory concentration (IC) ≈ 25 and 50 µg, respectively). By stabilization studies of topoisomerase I-DNA complex and preincubation studies of topoisomerase I and II α with the extract; we conclude that the possible mechanism of inhibition is both; 1) stabilization of covalent complex of topo I-DNA complex and 2) direct inhibition of the enzyme topoisomerases. These findings might explain the antineoplastic activity of Picrorrhiza kurroa and encourage new studies to elucidate the usefulness of the extract as a potent antineoplastic agent.

Keywords: Picrorrhiza kurroa, topoisomerase I and II α, inhibition, antineoplastic agent

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Authors: M. Djekoun, H. Berrebah, M. R. Djebar

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Fusarium wilt is destructive disease of cereal crops with small grains. It affects yields but also the quality of the crop and economic losses arising are often very heavy. Chemical control is currently one of the most effective ways to fight against these diseases. In this study, the efficacy of three fungicides (tebuconazole, thiram, and fludioxonil-difenoconazole mixture) was tested. In vitro, on the phytopathogenic Fusarium spp. isolated from seeds of wheat. The active ingredients were tested at different concentrations: 0.06, 1.39, 2.79, 5.58, and 11.16 mg/l for tebuconazole, 0.035, 0.052, 0.105, 0.21, and 0.42 mg/l for thiram and finally, for the mixture fludioxonil-difenoconazole 4 concentrations were tested: 0.05, 0.1, 0.5 and 1 mg/l. Toxicity responses were expressed as effective concentration, which inhibits mycelial growth by 50%, (EC50). Of the three selected fungicides, thirame proved to be the most effective with EC50 value of the order of 0,15 mg/l followed by the mixture of fludioxonil-difenoconazole with 0,27mg/l and finally tebuconazole with a value of 3.79 mg/l.

Keywords: Fusarium spp., thiram, tebuconazole, fludioxonil, difenoconazole, percentage of inhibition, EC50

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Authors: Ozlem Temiz-Arpaci, Meryem Tasci, Fatma Sezer Senol, İlkay Erdogan Orhan

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Alzheimer’s disease (AD), a neurodegenerative disorder characterized by a progressive deterioration of memory and cognition, occurs more frequently in elderly people. Current treatment approaches in this disease with the major therapeutic strategy are based on the AChE and BChE inhibition. On the other hand, tyrosinase inhibition has become a target for the treatment of Parkinson’s disease (PD) since this enzyme may play a role in neuromelanin formation in the human brain and could be critical in the formation of dopamine neurotoxicity associated with neurodegeneration linked to PD. Also benzoxazoles are structural isosteres of natural nucleotides that can interact with biopolymers so that benzoxazoles showed a lot of different biological activities. In this study, a series of 2,5-disubstituted-benzoxazole derivatives were synthesized and were evaluated as possible inhibitors of acetylcholinesterase (AChE) / butyrylcholinesterase (BChE) and tyrosinase. The results demonstrated that the compounds exhibited a weak spectrum of AChE / BChE inhibitory activity ranging between 3.92% - 54.32% except compound 8 which showed no activity against AChE and compound 4 which showed no activity against BChE at the specified molar concentrations. Also, the compounds indicated lower than tyrosinase inhibitory activity of ranging between 8.14% - 22.90% to that of reference (kojic acid).

Keywords: AChE and BChE inhibition, Alzheimer’s disease, benzoxazoles, tyrosinase inhibition

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Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: trichoderma spp., meloidogyne javanica, Fusarium oxysporum f.sp.radicis lycopersici, biocontrol

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Authors: Liang Jiansheng, Zhu Wei

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Light and brassinosteroids are essential external and internal cues for plant survival. Although the coordination of light with phytohormone signals is crucial for plant growth and development, the molecular connection between light and brassinosteroid signaling during root soil penetration remains elusive. Here, we reveal that light-stabilized RACK1 couples a brassinosteroid signaling cascade to drive cell division in root meristems. RACK1 family scaffold proteins positively regulate light-induced the promotion of root elongation during soil penetration. Under the light condition, RACK1A interacts with both phyB and SPA1, then reinforces the phyB-SPA1 association to accumulate its abundance in roots. In response to brassinosteroid signals, RACK1A competes with BKI1 to attenuate the BRI1-BKI1 interaction, thereby leading to activating BRI1 actions in root development. Furthermore, RACK1A binds to BES1 to repress its DNA binding activity toward the target gene CYCD3;1. This ultimately allows to release the inhibition of CYCD3;1 transcription, and promotes cell division during root growth. Our study illustrates a new mechanistic model of how plants engage scaffold proteins in transducing light information to facilitate brassinosteroid signaling for root growth in the soil.

Keywords: root growth, cell division, light signaling, brassinosteroid signaling, soil penetration, scaffold protein, RACK1

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Authors: C. Mekala, Indumathi M. Nambi

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Nitrogen pollution in groundwater arising from wastewater and fertilizer application through vadose zone is a major problem and it causes a prime risk to groundwater based drinking water supplies. Nitrogenous compounds namely ammonium, nitrate and nitrite fate and transport in soil subsurface were studied experimentally. The major process like sorption, leaching, biotransformation involving microbial growth kinetics, and biological clogging due to biomass growth were assessed and modeled with advection-dispersion reaction equations for ammonium, nitrate and acetate in a saturated, heterogeneous soil medium. The transport process was coupled with freundlich sorption and monod inhibition kinetics for immobile bacteria and permeability reduction due to biomass growth will be verified and validated with the numerical model. This proposed mathematical model will be very helpful in the development of a management model for a sustainable and safe wastewater reuse strategies such as irrigation and groundwater recharge.

Keywords: nitrogen species transport, transformation, biological clogging, biokinetic parameters, contaminant transport model, saturated soil

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Authors: Yu-Wen Cheng, Jau-Der Ho, Liang-Huan Wu, Fan-Li Lin, Li-Huei Chen, Hung-Ming Chang, Yueh-Hsiung Kuo, George Hsiao

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Retina glial activation and neuroinflammation have been confirmed to cause devastating responses in retinodegenerative diseases. The expression and activation of matrix metalloproteinase (MMP)-9 and inducible nitric oxide synthase (iNOS) could be exerted as the crucial pathological factors in glaucoma- and blue light-induced retinal injuries. The present study aimed to investigate the retinoprotective effects and mechanisms of fungal ingredient 3,4-dihydroxybenzalacetone (DBL) isolated from Phellinus linteus in the retinal glial activation and retinodegenerative animal models. According to the cellular studies, DBL significantly and concentration-dependently abrogated MMP-9 activation and expression in TNFα-stimulated retinal Müller (rMC-1) cells. We found the inhibitory activities of DBL were strongly through the STAT- and ERK-dependent pathways. Furthermore, DBL dramatically attenuated MMP-9 activation in the stimulated Müller cells exposed to conditioned media from LPS-stimulated microglia BV-2 cells. On the other hand, DBL strongly suppressed LPS-induced production of NO and ROS and expression of iNOS in microglia BV-2 cells. Consistently, the phosphorylation of STAT was substantially blocked by DBL in LPS-stimulated microglia BV-2 cells. In the evaluation of retinoprotective functions, the high IOP-induced scotopic electroretinographic (ERG) deficit and blue light-induced abnormal pupillary light response (PLR) were assessed. The deficit scotopic ERG responses markedly recovered by DBL in a rat model of glaucoma-like ischemia/reperfusion (I/R)-injury. DBL also reduced the aqueous gelatinolytic activity and retinal MMP-9 expression in high IOP-injured conditions. Additionally, DBL could restore the abnormal PLR and reduce retinal MMP-9 activation. In summary, DBL could ameliorate retinal neuroinflammation and MMP-9 activation by predominantly inhibiting STAT3 activation in the retinal Müller cells and microglia, which exhibits therapeutic potential for glaucoma and other retinal degenerative diseases.

Keywords: glaucoma, blue light, DBL, retinal Müller cell, MMP-9, STAT, Microglia, iNOS, ERG, PLR

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Authors: David Sarpong, Waleed Khursheed, Ernest Danquah, Erin Murphy

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Shigella is a pathogenic bacterium responsible for shigellosis, a severe diarrheal disease that claims the lives of immunocompromised individuals worldwide. To develop therapeutics against this disease, an understanding of the molecular mechanisms underlying the pathogen’s physiology is crucial. Small non-coding RNAs (sRNAs) have emerged as important regulators of bacterial physiology, including as components of toxin-antitoxin systems. In this study, we investigated the role of RyfA in S. flexneri physiology and virulence. RyfA, originally identified as an sRNA in Escherichia coli, is conserved within the Enterobacteriaceae family, including Shigella. Whereas two copies of ryfA are present in S. dysenteriae, all other Shigella species contain only one copy of the gene. Additionally, we identified a putative open reading frame within the RyfA transcript, suggesting that it may be a dual-functioning gene encoding a small protein in addition to its sRNA function. To study ryfA in vitro, we cloned the gene into an inducible plasmid and observed the effect on bacterial growth. Here, we report that RyfA production inhibits the growth of S. flexneri, and this inhibition is dependent on the contained open reading frame. In-silico analyses have revealed the presence of two divergently transcribed sRNAs, RyfB1 and RyfB2, which share nucleotide complementarity with RyfA and thus are predicted to function as anti-toxins. Our data demonstrate that RyfB2 has a stronger antitoxin effect than RyfB1. This regulatory pattern suggests a novel form of a toxin-antitoxin system in which the activity of a single toxin is inhibited to varying degrees by two sRNA antitoxins. Studies are ongoing to investigate the regulatory mechanism(s) of the antitoxin genes, as well as the downstream targets and mechanism of growth inhibition by the RyfA toxin. This study offers distinct insights into the regulatory mechanisms underlying Shigella physiology and may inform the development of new anti-Shigella therapeutics.

Keywords: sRNA, shigella, toxin-antitoxin, Type 1 toxin antitoxin

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Authors: Asad Ali, Asif Jamal

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The agriculture productivity losses due to microbial pathogens have been a serious issue in Pakistan and rest of the world. Present work was designed to isolate soil borne microorganisms having the antagonistic ability against notorious phytopathogens. From the initial collection of 23 bacterial isolates, two potent strains of Bacillus were screened on the basis of their comparative efficacy against devastating fungal pathogens. The strains AK-1 and AK-5 showed excellent inhibitory indexes against the majority of tested fungal strains. It was noted that both strains of Bacillus showed significant biocontrolling activity against Aspergillus flavus, Fusarium moniliforme, Colletotricum falcatum, Botrytis cinerea, Aspergillus niger, Fusarium oxysporum, Phythopthora capsici and Rhizopus oryzae. The strain AK-1 was efficient to suppress Aspergillus species and Rhizopus oryzae while AK-5 expressed significant antagonistic activity against Fusarium, Botrytis and Colletotricum species. On the basis of in vitro assay, it can be postulated that the Bacillus strains AK-1 and AK-5 can be used as bio-protective agent against various plant diseases. In addition, their applications as natural pesticides could be very helpful to prevent the adverse effects of chemical pesticides.

Keywords: bacillus species, biocontrol agent, biopesticides, phytopathogens

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Authors: Sarah Ullrich, Juliane Rolle, Christian Beste, Nicole Wolff

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Inhibition and cognitive flexibility are essential parts of executive functions in our daily lives, as they enable the avoidance of unwanted responses or selectively switch between mental processes to generate appropriate behavior. There is growing interest in improving inhibition and response selection through brief mindfulness-based meditations. Arguably, open-monitoring meditation (OMM) improves inhibitory and flexibility performance by optimizing cognitive control and information processing. Yet, the underlying neurophysiological processes have been poorly studied. Using the Simon-Go/Nogo paradigm, the present work examined the effect of a single 15-minute smartphone app-based OMM on inhibitory performance and response selection in meditation novices. We used both behavioral and neurophysiological measures (event-related potentials, ERPs) to investigate which subprocesses of response selection and inhibition are altered after OMM. The study was conducted in a randomized crossover design with N = 32 healthy adults. We thereby investigated Go and Nogo trials in the paradigm. The results show that as little as 15 minutes of OMM can improve response selection and inhibition at behavioral and neurophysiological levels. More specifically, OMM reduces the rate of false alarms, especially during Nogo trials regardless of congruency. It appears that OMM optimizes conflict processing and response inhibition compared to no meditation, also reflected in the ERP N2 and P3 time windows. The results may be explained by the meta control model, which argues in terms of a specific processing mode with increased flexibility and inclusive decision-making under OMM. Importantly, however, the effects of OMM were only evident when there was the prior experience with the task. It is likely that OMM provides more cognitive resources, as the amplitudes of these EKPs decreased. OMM novices seem to induce finer adjustments during conflict processing after familiarization with the task.

Keywords: EEG, inhibition, meditation, Simon Nogo

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Authors: N. Naga Subrahmanyeswara Rao, Parag Arvind Deshpande

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Understanding nucleotide synthesis reaction of any organism is beneficial to know the growth of it as in Mycobacterium tuberculosis to design anti TB drug. One of the reactions of de novo pathway which takes place in all organisms was considered. The reaction takes places between phosphoribosyl pyrophosphate and orotate catalyzed by orotate phosphoribosyl transferase and divalent metal ion gives orotdine monophosphate, a nucleotide. All the reaction steps of three experimentally proposed mechanisms for this reaction were considered to develop kinetic rate expression. The model was validated using the data for four organisms. This model could successfully describe the kinetics for the reported data. The developed model can serve as a reliable model to describe the kinetics in new organisms without the need of mechanistic determination. So an organism-independent model was developed.

Keywords: mechanism, nucleotide, organism, tuberculosis

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Authors: María Graciela Aguayo, Laura Reyes, Claudia Oviedo, José Navarrete, Liset Gómez, Hugo Torres

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Most wood species used in construction deteriorate when exposed to environmental conditions that favor wood-degrading organisms’ growth. Therefore, chemical protection by impregnation allows more efficient use of forest resources extending the wood useful life. A wood protection treatment which has attracted considerable interest in the scientific community during the last decade is wood impregnation with nano compounds. Radiata pine is the main wood species used in the Chilean construction industry, with total availability of 8 million m³ sawn timber. According to the requirements of the American Wood Protection Association (AWPA) and the Chilean Standards (NCh) radiata pine timber used in construction must be protected due to its low natural durability. In this work, the impregnation with copper nanoparticles (CuNP) was studied in terms of penetration and its protective effect against wood rot fungi. Two concentrations: 1 and 3 g/L of NPCu were applied by impregnation on radiata pine sapwood. Test penetration under AWPA A3-91 standard was carried out, and wood decay tests were performed according to EN 113, with slight modifications. The results of penetration for 1 g/L CuNP showed an irregular total penetration, and the samples impregnated with 3 g/L showed a total penetration with uniform concentration (blue color in all cross sections). The impregnation wood mass losses due to fungal exposure were significantly reduced, regardless of the concentration of the solution or the fungus. In impregnated wood samples, exposure to G. trabeum resulted ML values of 2.70% and 1.19% for 1 g/L and 3 g/L CuNP, respectively, and exposure to P. placenta resulted in 4.02% and 0.70%-ML values for 1 g/L and 3 g/L CuNP, respectively. In this study, the penetration analysis confirmed a uniform distribution inside the wood, and both concentrations were effective against the tested fungi, giving mass loss values lower than 5%. Therefore, future research in wood preservatives should focus on new nanomaterials that are more efficient and environmentally friendly. Acknowledgments: CONICYT FONDEF IDeA I+D 2019, grant number ID19I10122.

Keywords: copper nanoparticles, fungal degradation, radiata pine wood, wood preservation

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Authors: Shivankar Agrawal, Indira Sarangthem

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Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA) remains one of the major causes of healthcare-associated and community-onset infections worldwide. Hence the search for non-toxic natural compounds having antibacterial activity has intensified for future drug development. The exploration of less studied niches of Earth can highly increase the possibility to discover novel bioactive compounds. Therefore, in this study, the cultivable fraction of fungi from the sediments of natural hot springs has been studied to mine potential fungal candidates with antibacterial activity against the human pathogen Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. We isolated diverse strains of thermophilic fungi from a collection of samples from sediment. Following a standard method, we isolated a promising thermophilic fungus strain IBSD19, identified as Acrophialophora levis, possessing the potential to produce an anti-Staphylococcus aureus agent. The growth conditions were optimized and scaled to fermentation, and its produced extract was subjected to chemical extraction. The ethyl acetate fraction was found to display significant activity against Staphylococcus aureus and MRSA with a minimum inhibitory concentration (MIC) of 0.5 mg/ml and 4 mg/ml, respectively. The cell membrane integrity assay and SEM suggested that the fungal metabolites cause bacteria clustering and further lysis of the cell.

Keywords: antibacterial activity, antioxidant, fungi, Staphylococcus aureus, MRSA, thermophiles

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Authors: Ming-Tzu Tsai, Jui-Ting Hsu, Chia-Chieh Lin, Feng-Tsai Chiang, Cheng-Chin Huang

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Far infrared (FIR), an invisible and short electromagnetic waves ranges from 6-14 μm also defines as the “growth ray.” Although the mechanism of FIR is still unknown, most data have suggested that FIR could accelerate the skin microcirculation by elevating the blood flow and nitric-oxide (NO) synthesis. In this present work, the effect of FIR irradiation and endothelial cell growth supplement (ECGS) on human umbilical vascular endothelial cells (HUVECs) was evaluated. To understand whether the cell viability and NO production of HUVECs affected by NO, cells with/without ECGS were treated in the presence or absence of L-NAME, an eNOS inhibitor. For FIR exposure, FIR-emitted ceramic powders consisted of a variety of well-mixed metal oxides were developed. The results showed that L-NAME did had a strong effect on the inhibition of NO production, especially in the ECGS-treated group. However, the cell viability of each group was rarely affected in the presence of L-NAME. Cells with the incubation of ECGS showed much higher cell viability compared to the control. Moreover, NO production of HUVECs exposed to FIR irradiation was significantly inhibited in the presence of L-NAME. It suggested that NO could play a role modulating the downstream signals of HUVECs during FIR exposure.

Keywords: far-infrared irradiation (FIR), nitric oxide (NO), endothelial nitric oxide synthase (eNOS), endothelial cell growth supplement (ECGS)

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Authors: Riccardo Cacciotti, Jiri Frankl, Benjamin Wolf, Michael Machacek

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Moisture fluctuations govern the occurrence of fungi-related problems in buildings, which may impose significant health risks for users and even lead to structural failures. Several numerical engineering models attempt to capture the complexity of mold growth on building materials. From real life observations, in cases with suppressed daily variations of boundary conditions, e.g. in crawlspaces, mold growth model predictions well correspond with the observed mold growth. On the other hand, in cases with substantial diurnal variations of boundary conditions, e.g. in the ventilated cavity of a cold flat roof, mold growth predicted by the models is significantly overestimated. This study, founded by the Grant Agency of the Czech Republic (GAČR 20-12941S), aims at gaining a better understanding of mold growth behavior on solid wood, under varying boundary conditions. In particular, the experimental investigation focuses on the response of mold to changing conditions in the boundary layer and its influence on heat and moisture transfer across the surface. The main results include the design and construction at the facilities of ITAM (Prague, Czech Republic) of an innovative device allowing for the simulation of changing environmental conditions in buildings. It consists of a square section closed circuit with rough dimensions 200 × 180 cm and cross section roughly 30 × 30 cm. The circuit is thermally insulated and equipped with an electric fan to control air flow inside the tunnel, a heat and humidity exchange unit to control the internal RH and variations in temperature. Several measuring points, including an anemometer, temperature and humidity sensor, a loading cell in the test section for recording mass changes, are provided to monitor the variations of parameters during the experiments. The research is ongoing and it is expected to provide the final results of the experimental investigation at the end of 2022.

Keywords: moisture, mold growth, testing, wood

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Authors: Vanja Vlajkov, Ivana PajčIn, Mila Grahovac, Marta Loc, Dragana Budakov, Jovana Grahovac

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The rhizosphere soil refers to the plant roots' dynamic environment characterized by their inhabitants' high biological activity. Rhizospheric bacteria are recognized as effective biocontrol agents and considered cardinal in alternative strategies for securing ecological plant diseases management. The need to suppress fungal pathogens is an urgent task, not only because of the direct economic losses caused by infection but also due to their ability to produce mycotoxins with harmful effects on human health. Aspergillus and Fusarium species are well-known producers of toxigenic metabolites with a high capacity to colonize crops and enter the food chain. The bacteria belonging to the Bacillus genus has been conceded as a plant beneficial species in agricultural practice and identified as plant growth-promoting rhizobacteria (PGPR). Besides incontestable potential, the full commercialization of microbial biopesticides is in the preliminary phase. Thus, there is a constant need for estimating the suitability of novel strains to be used as a central point of viable bioprocess leading to market-ready product development. In the present study, 76 potential producing strains were isolated from the rhizosphere soil, sampled from different localities in the Autonomous Province of Vojvodina, Republic of Serbia. The selective isolation process of strains started by resuspending 1 g of soil samples in 9 ml of saline and incubating at 28° C for 15 minutes at 150 rpm. After homogenization, thermal treatment at 100° C for 7 minutes was performed. Dilution series (10-1-10-3) were prepared, and 500 µl of each was inoculated on nutrient agar plates and incubated at 28° C for 48 h. The pure cultures of morphologically different strains indicating belonging to the Bacillus genus were obtained by the spread-plate technique. The cultivation of the isolated strains was carried out in an Erlenmeyer flask for 96 h, at 28 °C, 170 rpm. The antagonistic activity screening included two phytopathogenic fungi as test microorganisms: Aspergillus sp. and Fusarium sp. The mycelial growth inhibition was estimated based on the antimicrobial activity testing of cultivation broth by the diffusion method. For the Aspergillus sp., the highest antifungal activity was recorded for the isolates Kro-4a and Mah-1a. In contrast, for the Fusarium sp., following 15 isolates exhibited the highest antagonistic effect Par-1, Par-2, Par-3, Par-4, Kup-4, Paš-1b, Pap-3, Kro-2, Kro-3a, Kro-3b, Kra-1a, Kra-1b, Šar-1, Šar-2b and Šar-4. One-way ANOVA was performed to determine the antagonists' effect statistical significance on inhibition zone diameter. Duncan's multiple range test was conducted to define homogenous groups of antagonists with the same level of statistical significance regarding their effect on antimicrobial activity of the tested cultivation broth against tested pathogens. The study results have pointed out the significant in vitro potential of the isolated strains to be used as biocontrol agents for the suppression of the tested mycotoxigenic fungi. Further research should include the identification and detailed characterization of the most promising isolates and mode of action of the selected strains as biocontrol agents. The following research should also involve bioprocess optimization steps to fully reach the selected strains' potential as microbial biopesticides and design cost-effective biotechnological production.

Keywords: Bacillus, biocontrol, bioprocess, mycotoxigenic fungi

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Authors: A. Majeed, M.K. Abbasi, S. Hameed, A. Imran, T. Naqqash, M. K. Hanif

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Plant growth-promoting rhizobacteria (PGPR) are free-living soil bacteria that aggressively colonize the rhizosphere/plant roots, and enhance the growth and yield of plants when applied to seed or crops. Root associated (endophytic and rhizospheric) PGPR were isolated from Sunflower (Helianthus anus) grown in soils collected from 16 different sites of sub division Dhirkot, Poonch, Azad Jammu & Kashmir, Pakistan. A total of 150 bacterial isolates were isolated, purified, screened in vitro for their plant growth promoting (PGP) characteristics. 11 most effective isolates were selected on the basis of biochemical assays (nitrogen fixation, phosphate solubilization, growth hormone production, biocontrol assay, and carbon substrates utilization assay through gas chromatography (GCMS), spectrophotometry, high performance liquid chromatography HPLC, fungal and bacterial dual plate assay and BIOLOG GN2/GP2 microplate assay respectively) and were tested on the crop under controlled and field conditions. From the inoculation assay, the most promising 4 strains (on the basis of increased root/shoot weight, root/shoot length, seed oil content, and seed yield) were than selected for colonization studies through confocal laser scanning and transmission electron microscope. 16Sr RNA gene analysis showed that these bacterial isolates belong to Pseudononas, Enterobacter, Azospirrilum, and Citobacter genera. This study is the clear evident that such isolates have the potential for application as inoculants adapted to poor soils and local crops to minimize the chemical fertilizers harmful for soil and environment

Keywords: PGPR, nitrogen fixation, phosphate solubilization, colonization

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