The widespread use of antibiotics in intensive animal husbandry, and the agricultural utilization of manure from such farms, imposes a significant burden on the environment. Consequently, the effects of antibiotics should be studied not only in animals and humans but also in all components of biocenoses and agrocenoses. In our study, we analyze the impact of four different concentrations of tetracycline present in soil (0, 5, 50, and 500 mg/kg of soil) on the growth and key photosynthesis parameters of pea seedlings: chlorophyll concentration, aminolevulinic acid concentration, aminolevulinic acid dehydrogenase activity, and ribulose bisphosphate carboxylase-oxygenase (RuBisCO) activity. At the lowest tetracycline concentration, chlorophyll content decreased by 13% compared to the control (0 tetracycline), while at the highest antibiotic concentration, it decreased by as much as 27%. Similarly, the decrease in aminolevulinic acid (a chlorophyll precursor) concentration was significant, amounting to 34%. However, the activity of the dehydrogenase enzyme, which consumes this precursor, decreased even more drastically by 51%, indicating significant disturbances in the light phase of photosynthesis. However, the activity of RuBisCO in pea plants subjected to tetracycline was even more severely affected, dropping by 58%, 69%, and 70% in soils with increasing concentrations of tetracycline. The reduction in enzyme activity could only partially be explained by a less pronounced decrease in the quantity of RuBisCO (large subunit) protein, which amounted to 6.5%, 11%, and 35% for tetracycline concentrations of 5, 50, and 500 mg/kg of soil, respectively.

The impact of tetracycline on the soil and rhizosphere microbiome of lettuce was analyzed. Soil was collected from an agricultural field regularly fertilized with manure, and tetracycline was added at two concentrations (5 mg/kg and 25 mg/kg). In untreated soil, dominant bacteria included Proteobacteria (43.17%), Bacteroidota (17.91%), and Firmicutes (3.06%). Tetracycline addition caused significant shifts in the microbiome composition, notably increasing Actinobacteriota (22%) and favoring Mycobacterium tuberculosis (low concentration) and Mycobacterium holsaticum (high concentration). Proteobacteria decreased by 21%, possibly indicating antibiotic resistance development. An increase in Firmicutes, particularly Bacillales, suggested a selection for resistant strains. In the lettuce rhizosphere, tetracycline-induced changes were less pronounced than in soil. Proteobacteria remained dominant, but taxa like Burkholderiales and Chitinophagales increased in response to tetracycline. The rise in chitin-degrading bacteria might result from fungal overgrowth linked to the bacteriostatic effects of tetracycline. Pathogens such as M. tuberculosis, observed in the soil, were not detected in the lettuce rhizosphere.

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.

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.