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Developing an effective method for isolating bacterial genetic material from plants is a relatively challenging task and often does not yield adequately prepared material for further analyses. Previous studies often overlook connections, primarily focusing on laboratory investigations. With advancements in high-throughput sequencing techniques, we can now revisit and delve deeper into these interactions. Our study focuses on the initial phase of these investigations: genetic material isolation. Extracting bacterial DNA from aboveground plant parts, known as the phyllosphere, poses a significant challenge due to plant-derived contaminants. Existing isolation protocols frequently yield inconsistent results, necessitating continuous refinement and optimization. In our study, we developed an effective isolation protocol employing mechanical–chemical lysis, sonication, and membrane filtration. This approach yielded high-quality DNA at a concentration of 38.08 ng/µL, suitable for advanced sequencing applications. Our results underscore the effectiveness and necessity of these methods for conducting comprehensive microbiological analyses. Furthermore, our research not only lays the groundwork for further studies on lettuce microbiota, but also highlights the potential for utilizing our developed protocol in investigating other plants and their microbiomes. [ABSTRACT FROM AUTHOR]

The impact of fluorine on plants remains poorly understood. We examined duckweed growth in extracts of soil contaminated with fluorine leached from chicken manure. Additionally, fluorine levels were analyzed in fresh manure, outdoor-stored manure, and soil samples at varying distances from the manure pile. Fresh manure contained 37–48 mg F− × kg−1, while soil extracts contained 2.1 to 4.9 mg F− × kg−1. We evaluated the physiological effects of fluorine on duckweed cultured on soil extracts or in 50% Murashige–Skoog (MS) medium supplemented with fluorine concentrations matching those in soil samples (2.1 to 4.9 mg F− × L−1), as well as at 0, 4, and 210 mg × L−1. Duckweed exposed to fluorine displayed similar toxicity symptoms whether in soil extracts or supplemented medium. Fluoride at concentrations of 2.1 to 4.9 mg F− × L−1 reduced the intact chlorophyll content, binding the porphyrin ring at position 32 without affecting Mg2+. This reaction resulted in chlorophyll a absorption peak shifted towards shorter wavelengths and formation of a new band of the F−-chlorophyll a complex at λ = 421 nm. Moreover, plants exposed to low concentrations of fluorine exhibited increased activities of aminolevulinic acid dehydratase and chlorophyllase, whereas the activities of both enzymes sharply declined when the fluoride concentration exceeded 4.9 mg × L−1. Consequently, fluorine damages chlorophyll a, disrupts the activity of chlorophyll-metabolizing enzymes, and diminishes the plant growth rate, even when the effects of these disruptions are too subtle to be discerned by the naked human eye. [ABSTRACT FROM AUTHOR]

Intensive poultry production may result in substantial emissions of pollutants into the environment, including pharmaceuticals and other chemicals used in poultry farming. The objective of this study was to verify the presence of ciprofloxacin, enrofloxacin, carbamazepine, metoclopramide, trimethoprim, diflufenican, flufenacet, and p,p′-DDE in soil and water in the immediate vicinity of a poultry manure heap. The influence of soil contaminants on the growth and selected physiological parameters of seed peas and common duckweed (as indicator plants) was tested. It has been proven that the cultivation of pea plants on soil coming from the close proximity of a heap of manure results in a deterioration of both morphological parameters (root length, shoot length) and physiological parameters (chlorophyll absorption, aminolevulinic acid dehydrogenase (ALAD) activity, aminolevulinic acid (ALA) content, lipid peroxidation, mitochondrial damage or production of HSP70 proteins). Similarly, water extracts from cultivated soils had a significant effect on duckweed, and it was found that contaminant leachates are indeed detectable in soil, groundwater, and deep water. Special attention should, therefore, be paid to the location, methods of storage, and use of poultry fertilizer. [ABSTRACT FROM AUTHOR]

Antibiotics as environmental pollutants can affect both the morphology and physiology of higher plants. The first visible symptoms of damage caused by antibiotics are chlorotic changes resulting from the degradation of chlorophyll and the impairment of the chlorophyll biosynthesis pathway, followed by damage to PS II and disturbances in the electron flow between the photosystems. Research on the effects of antibiotics, on plants has recently intensified, however, our understanding of this problem is still far from complete. This paper provides a review of current data on the impacts antibiotics on plant photosynthetic apparatus. [ABSTRACT FROM AUTHOR]

With increasing soil concentrations of ciprofloxacin and tetracycline a decrease of leaf chlorophyll content was observed. Tetracycline was more detrimental than ciprofloxacin. The chlorophyll content in plants growing for ten days on a tetracycline containing soil decreased by 68%. The decrease of chlorophyll concentration was even sharper in new leaves that formed after application of the antibiotic (up to 81% drop). The comparison of absorption spectra of commercial, reagent grade chlorophyll, alone and incubated with antibiotics, has shown that ciprofloxacin and tetracycline can react directly with chlorophyll and decrease its concentration by 47.7% and 48.5%, respectively. The changes in fluorescence spectra confirmed the formation of chlorophyll degradation product. The chlorophyll decay was a second order reaction and depended on antibiotic concentration and duration of exposure. Reaction rate constants differed with antibiotics and their soil concentrations. With increasing contents of antibiotics in soil the constant of chlorophyll degradation rate in lupin plants increased from k = 870 M −1 day −1 for 3 mg ciprofloxacin to k = 2490 M −1 day −1 for 90 mg ciprofloxacin, and in the case of tetracycline the reaction rate constant increased from k = 1330 M −1 day −1 to k = 2910 M −1 day −1 . The sensitivity of chlorophyll to ciprofloxacin and tetracycline was confirmed by determining EC and TU indices. [ABSTRACT FROM AUTHOR]

Pharmaceuticals have become an emerging environmental protection problem due to their presence in water and soil ecosystems. The aim of this study was to determine the effects of tetracycline (TC), oxytetracycline (OTC) and chlortetracycline (CTC) (at concentrations from 0 to 8 mg kg-1 of soil) on peroxidase activity in pea roots. Furthermore, the influence of tetracycline on germination and elongation of pea root was examined. The content of soluble carbohydrates in seedlings of pea plants was also analyzed as indicator of their metabolic activity. The obtained results show that all TC, OTC and CTC (at concentrations above 0.400 mg kg-1) decreased the activity of peroxidase. Plants took up all the analyzed drugs, but the accumulation of CTC was lowest, and it was least toxic to peas. The most readily taken up drug was OTC, and its level in plant tissues amounted to 31 to 146 μg/g of fresh mass. [ABSTRACT FROM AUTHOR]

Copyright of Zeszyty Naukowe Uniwersytetu Przyrodniczego we Wrocawiu: Rolnictwo is the property of Wroclaw University of Environmental & Life Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Zeszyty Naukowe Uniwersytetu Przyrodniczego we Wrocawiu: Rolnictwo is the property of Wroclaw University of Environmental & Life Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Zeszyty Naukowe Uniwersytetu Przyrodniczego we Wrocawiu: Rolnictwo is the property of Wroclaw University of Environmental & Life Sciences and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Pharmaceuticals have become an emerging environmental protection problem due to their presence in water and soil ecosystems. Reliable cell viability biomarkers (mitochondrial and cytosol distribution of cytochrome c oxidase activity), germination, and growth of seedlings were assessed to be sensitive endpoints of diclofenac toxicity. Moreover, the content of soluble carbohydrates in seedlings of three leguminous plants was an additional indicator of germination. The tested diclofenac concentrations (from 0 to 12 mM) in the three plant species (lupin, pea, and lentil) resulted in increased activity of the enzyme in cytosol, and a decreased activity in mitochondrions. The increase of the cytochrome c oxydase activity in cytosol was most rapid in pea and slowest in lupin. The decrease in mitochondrions was gradual, yet in roots growing in the soil contaminated with 12 mM of diclofenac, from 35 to 68% of total enzyme activity leaked from the mitochondrion to the cytoplasm. The dynamics of seedling growth was a better parameter of soil contamination with diclofenac than germination. On the basis of the described morphological and biochemical features, it was found that diclofenac is decidedly less phytotoxic toward leguminous plants (lupin, pea, lentil) than e.g. sulfamethazine. The research has shown that carbohydrate metabolism is a good parameter of seedling growth, but it is not an indicator of contamination and thus cannot be applied to assess soil ecosystem contamination with medicines. [ABSTRACT FROM AUTHOR]

Anthelmintic compounds are widely used in the world against endo- and ecto-parasites in medicine and veterinary medicine. Several groups of antiparasites are used comprising avermectins and benzimidazoles. Avermectins due to their high potency and wide spectrum of activity are some of the most potent compounds and they are also used as insecticides. The goal of the present study was the evaluation of the sensitivity of culture-free microbiotests to selected anthelmiotic pharmaceuticals. Four tests were applied with aquatic test biota: bacteria, protozoa, rotifera and crustacea. Moreover, a direct contact test for freshwater sediments (Ostracodtoxkit F™) was utilized. In this assay a freshwater benthic ostracod crustacean Heterocypris incongruens freshly hatched from cysts is used as a test biota. Seven pharmaceuticals were tested: avermectins (abamectin, ivermectin, doramectin), benzimidazoles (albendazole, mebendazole, thiabendazole) and morantel. Avermectins were not toxic to luminescent bacteria Vibrio fisheri and protozoan Spirostomum ambiguum, but they were toxic to rotifer Brachionus calyciflorus, with 24h-LC50 values from 1.57 to 2.38 mg l-1. The highest toxicity was observed for crustaceans, both planktonic Thamnocephalus platyurus and benthic Heterocypris incongruens. LC50 values ranged from 5.7 to 25.2 μg l-1. Morantel did not cause adverse effects in any microbiotests at concentrations up to 10mg l-1. [ABSTRACT FROM AUTHOR]

The aim of this study was to compare the physiological responses of six plant species (popular crops or plants recommended as indicators of soil pollution) to a wide range of glyphosate concentrations (0, 1, 3, 7, 10, 40, 80, 120, 180, 240, 400, 750, 1,000, 1,500, 1,700 and 2,000 μM). Percent germination, root length, seedling dry mass and myo-inositol content, as well as seedling leachate electroconductivity were determined in Lepidium sativum, Sinapis alba, Sorghum saccharatum, Brassica napus, Lupinus luteus and Avena sativa. Percent seed germination, seedling dry mass and electroconductivity of seedling leachates were not clearly affected by the herbicide and could not be used as indicators of its phytotoxicity. An metabolite induced by abiotic stresses in many plants, myo-Inositol, was very strongly stimulated by glyphosate at doses above 10 or 40 μM, depending on plant species. The sensitivity of analyzed plants to glyphosate, as manifested by root length, differed clearly. In Avena sativa the relationship between root length and glyphosate concentration was fairly linear over a wide range of herbicide doses (up to 240-400 μM). The most distinct drop in root growth at low herbicide doses was visible in Sorghum saccharatum. The results show that a mild stress affecting root length may not clearly modify seedling myo-inositol levels, that respond distinctly to stronger stresses. Not all indicator plants are equally suitable for analysis of biological activity of glyphosate residues. Sorghum saccharatum seems particularly sensitive. [ABSTRACT FROM AUTHOR]

Drugs used in human and veterinary medicine belong to a new class of environmental pollutants. Antibiotics and chemotherapeutics may produce biological effects at very low concentrations. Moreover, the continuous exposure of bacteria to even small concentrations of antibiotics or active metabolites could lead to the emergence of resistant bacteria strains. Antibiotics may also have high biological activity against non-target organisms as plants. Biotests, opposite of instrumental (chemical) methods, allow estimation of whether very low levels of active substance residues in the soil can be phytotoxic to crop plants. The aim of this study was to evaluate the biogenic amines content in lupin seedlings as affected by different enrofloxacin concentrations in soil. With increasing enrofloxacin concentrations the root growth was inhibited more severely and dry mass increased slightly but steadily. At the highest enrofloxacin concentration, the dry mass of both roots and stems did not exceed 15% fresh mass. The lowest content of raffinose family oligosaccharides (RFOs) was observed in seedlings growing in the soil without enrofloxacin, the highest in those growing in soil containing 50 mM of the antibiotic. The speed of RFO mobilization during germination was a good indicator of elongation in seedlings. A lowered RFO content during germination was correlated with an increase in glucose, galactose, and sucrose content. Spermine, spermidine, and putrescyne were detected in roots. The total biogenic amine content ranged 7-105 µM·g-1 fresh mass. The results showed, that the biogenic amine profiles seem to be good parameters for the qualification of plant products. [ABSTRACT FROM AUTHOR]