Polycyclic aromatic hydrocarbons (PAHs) are hazardous organic compounds with mutagenic, genotoxic and carcinogenic properties. Although PAHs in soil can cause toxicity to microorganisms, the microbial community is able to degrade these compounds. For this reason, it is important to study acute and short-term effects of PAH contamination on soil microbial community, also to shed light on its possible exploitation in soil restoration.The effects of acute PAH contamination on the structure and metabolic activity of microbial communities in three forest (beech, holm oak, black pine) soils were studied. The soils were spiked with phenanthrene, pyrene or benzo[a]pyrene and incubated in experimental mesocosms, under controlled conditions. Enzymatic activities (laccase, total peroxidase and hydrolase), as well as microbial biomass and community structure (through phospholipid fatty acid and ergosterol analyses), were evaluated in the three soil systems 4 days after contamination and compared to no-spiked soils. In soil under holm oak, there was a stimulation of Gram+ bacteria after contamination with all the 3 PAHs, whereas in soil under pine, pyrene and phenanthrene additions mainly stimulated fungi and actinomycetes.

Tango® Super is a two-compound fungicide formulation widely employed in grain protection. However, details of Tango® Super effects on cell cultures have not been fully investigated. In this study, bovine lymphocytes were exposed to a concentration range 0.5; 1.5; 3; 6; and 15 μg mL⁻¹ for 4 h to assess the cytotoxicity and genotoxicity of the fungicide. Our experiments revealed that this fungicide treatment reduced cell viability, decreased cell proliferation and provoked apoptotic cell death. Cell cycle analysis showed predominant accumulation of cells in the G0/G1 phase of the cell cycle. The fungicide was able to induce mitochondrial superoxide production accompanied by elevated levels of carbonylated proteins and changes in the lipid membrane composition. The fungicide did not induce micronuclei production, but stimulated both DNA double-strand breaks and the formation of p53 binding protein, which is accumulated during the DNA repair process at the site of double-strand breaks. Based on the obtained data we suppose that the fungicide-induced DNA damage is the result of oxidative stress, which may contribute to higher occurrence of apoptotic cell death. Because ergosterol biosynthesis-inhibiting fungicides are widely used in agriculture to ensure higher crop yields and may cause health impairment of animals and humans, there is a need for further testing to elucidate their potential genotoxic effects using in vivo and/or in vitro systems.

Serious environmental pollution of heavy metals has attracted people's attention in recent years and halophiles seem to be potential bioremediation in the controlling of heavy metals contamination. In this study, the adaptive mechanism of halophilic Brachybacterium muris (B. muris) in response to salt stress and its mitigation of copper (Cu) toxicity in hydroponic plants were investigated. The cell morphology was observed using transmission electron microscopy. The cell membrane composition and fluidity were examined by the combination of gas chromatography, gas chromatography-mass spectrometry, ultra-high performance liquid chromatography-mass spectrometry, and fluorescence spectrophotometry. Moreover, the metabolic pathways of B. muris in response to salt stress were analyzed using the prokaryotic transcriptomics approach. A hydroponic co-culture model was further conducted to explore the effects of B. muris on wheat seedlings subjected to Cu toxicity. It was found that B. muris can respond to high osmotic pressure by improving the cell membrane fluidity, altering the cell morphology and cell membrane compositions. The proportion of unsaturated fatty acids, phosphatidylethanolamine, and phosphatidylinositol in B. muris cell membranes increased significantly, while zymosterol, fecosterol, and ergosterol contents decreased under a high salinity situation. Further transcriptomic analysis showed that genes encoding L-glutamate synthase, glutamate ABC transporter ATP-binding protein, and sodium cotransporter were up-regulated, indicating that both the synthesis and transport of glutamate were significantly enhanced under high osmotic pressure. Additionally, B. muris alleviated the inhibitory effect of Cu²⁺ on wheat seedlings' growth, causing a 30.14% decrease in H₂O₂ content and a significant increase of 83.86% and 45.96% in POD activity and GSH content in wheat roots, respectively. The findings of this study suggested that the salt-tolerant B. muris may serve as a promising strategy for improving the bioremediation of metal-contaminated saline water and soils.

Azoles are effective antifungal agents used in both medicine and agriculture. They typically work by inhibiting cytochrome P450 enzymes, primarily CYP51 of the ergosterol biosynthesis pathway, thus damaging the fungal cell membrane. However, apart from their desired antifungal properties, several azoles also exhibit endocrine disrupting properties in mammals, both in vitro and in vivo. Here, we have tested two currently used agricultural azole fungicides, triticonazole and flusilazole, for their in vitro anti-androgenic activity and potential effects on reproductive parameters. Both fungicides showed strong androgen receptor (AR) antagonism and disruption of steroid biosynthesis in vitro. Following gestational exposure to flusilazole (15 or 45 mg/kg bw/day) or triticonazole (150 or 450 mg/kg bw/day) in time-mated Sprague Dawley rats, triticonazole induced shorter male anogenital distance (AGD). Flusilazole exposure did not affect the AGD, but altered fetal male blood hormone profile, with increased androstenedione and decreased estrone levels. Flusilazole and triticonazole have dissimilar effects on reproductive parameters in vivo, but both show endocrine disrupting activities.

Sewage pollution is a principal factor of decreasing water quality, although it has not been considered a real impact in Amazonia that is still considered a pristine environment around the world. Thus, this study aimed to assess the levels of sewage contamination in sediments from three streams crossing Manaus − a Brazilian city of 2,403,796 inhabitants in the heart of the Amazon rain forest. Cholesterol, cholestanol, brassicasterol, ergosterol, stigmasterol, β-sitosterol, campesterol, stigmastanol, coprostanol, and epicoprostanol levels were determined by liquid chromatography tandem mass spectrometry (LC−MS/MS). The fecal indicator, coprostanol, was found in high concentrations (509−12 830 ng g⁻¹) and high relative proportions (21–54%) in all samples collected in the Mindu stream that crosses many heavily populated districts of the city, and in the Quarenta stream that crosses the Industrial District of Manaus. The sediments of the Tarumã-Açu stream also presented coprostanol; however, concentrations (<LOQ−142 ng g⁻¹) and relative proportions (0–7%) were much lower in this stream. Sterol ratios indicate a severe contamination of the urban streams (Mindu and Quarenta) and a low to moderate contamination of the partially urban stream (Tarumã-Açu). This is the first study evaluating the levels of sewage contamination of Amazon streams using sterol biomarkers and the results obtained herein indicate the need of an immediate implementation of effective sewage treatment strategies. Additionally, these findings may be considered as baseline concentrations for future monitoring programs of that globally important environment.

Present study deals with the efficacy of nanoencapsulated Homalomena aromatica essential oil (HAEO) as a potent green preservative against toxigenic Aspergillus flavus strain (AF-LHP-NS 7), storage fungi, AFB₁, and free radical-mediated deterioration of stored spices. GC–MS analysis revealed linalool (68.51%) as the major component of HAEO. HAEO was encapsulated into chitosan nanomatrix (CS-HAEO-Ne) and characterized through SEM, FTIR, and XRD. CS-HAEO-Ne completely inhibited A. flavus growth and AFB₁ biosynthesis at 1.25 μL/mL and 1.0 μL/mL, respectively in comparison to unencapsulated HAEO (1.75 μL/mL and 1.25 μL/mL, respectively). CS-HAEO-Ne caused significant reduction in ergosterol content in treated A. flavus and provoked leakage of cellular ions (Ca⁺², Mg⁺², and K⁺) as well as 260 nm and 280 nm absorbing materials. Depletion of methylglyoxal level in treated A. flavus cells illustrated the novel antiaflatoxigenic efficacy of CS-HAEO-Ne. CS-HAEO-Ne exhibited superior antioxidant efficacy (IC₅₀ ₍DPPH₎ = 4.5 μL/mL) over unencapsulated HAEO (IC₅₀ ₍DPPH₎ = 15.9 μL/mL) and phenolic content. CS-HAEO-Ne depicted excellent in situ efficacy by inhibiting fungal infestation, AFB₁ contamination, lipid peroxidation, and mineral loss with acceptable sensorial profile. Moreover, broad safety paradigm (LD₅₀ value = 7150.11 mg/kg) of CS-HAEO-Ne also suggests its application as novel green preservative to enhance shelf life of stored spices.

Although differential sensitivity of lichens to calcium excess has been documented previously at the community level, ecophysiological studies, which would shed light on the calcifuge or calcicole nature of lichens, are virtually absent. In the present study, we compared physiological responses of two morphologically similar foliose lichens, Dermatocarpon miniatum (calcicole lichen) and Umbilicaria hirsuta (calcifuge lichen) to Ca excess (up to 100 mM). The degree of total Ca uptake by the lichens after 24-h prolonged exposure was compared with selected physiological markers including levels of assimilation pigments, chlorophyll a fluorescence, soluble proteins, ergosterol, TBARS, and hydrogen peroxide. Both tested lichens accumulated Ca from the applied solutes of CaCl₂ by a dose-dependent manner, although excess of Ca did not change content of assimilation pigments in both tested lichens, as well as integrity of lichen symbiont membranes (tested as TBARS, K content, and ergosterol content) when compared to respective controls. However, we observed significant, concentration-dependent decrease of chlorophyll a fluorescence, content of soluble proteins, and hydrogen peroxide production in U. hirsuta, while in D. miniatum were all these parameters stable through all tested Ca concentrations.

Lichens absorb water, gases, dissolved substances, and especially pollutants by the entire surface, and they are considered to be the indicators of air quality. In our experiment, a sensitivity of Cladonia arbuscula subsp. mitis and Cladonia furcata lichens with the same photobiont Trebouxia was tested to nitrogen excess through a sensitivity of both the photobiont and mycobiont. Lichen ecophysiological parameters like chlorophyll a fluorescence, chlorophyll a integrity, the content of photosynthetic pigments, ergosterol, soluble proteins, thiobarbituric acid reactive substances, and secondary compounds were measured during two experiments that differed in time of nitrogen exposure. In the short-term experiment, also higher nitrogen concentrations were used to evaluate the dependence of different nitrogen concentrations. In the short-term experiment, lichens were soaked at the different solutions of ammonium nitrate (NH₄NO₃) for describing an immediate effect of range of NH₄NO₃ concentrations. In the long-term experiment, lichens were sprayed with low NH₄NO₃ concentrations for 3 months for evaluating the effect of naturally occurring low nitrogen concentrations. Results showed that lichens responded differently in spite of having the same photobiont. The mycobiont of C. arbuscula subsp. mitis was more sensitive than mycobiont of C. furcata. In higher nitrogen concentrations, the photobiont of C. furcata was more sensitive than C. arbuscula subsp. mitis photobiont. Both lichens exhibited signs of damage; therefore, we conclude that they are sensitive to nitrogen excess, while C. arbuscula subsp. mitis is more sensitive species. The secondary compound content did not change in neither of lichen species. Cladonia sp. response to nitrogen excess depends on length and nitrogen dose exposure.

The study reports the preservative efficacy of Bunium persicum (Boiss) essential oil (BPEO) against fungal and aflatoxin B₁ (AFB₁) contamination of stored masticatories and boosting of its efficacy through encapsulation into chitosan. BPEO was chemically characterized through GC-MS analysis, which revealed γ-terpinene as the major compound. The BPEO at 1.2 μL/mL concentration completely inhibited the growth of toxigenic strain of Aspergillus flavus (AF-LHP-PE-4) along with 15 common food borne moulds and AFB₁ secretion. The BPEO exerts its antifungal action on plasma membrane, as confirmed through ergosterol inhibition, alteration of membrane fluidity and enhancement of cellular ions and 260 and 280 nm absorbing material leakage. The antiaflatoxigenic mechanism of action of BPEO was confirmed through methylglyoxal reduction. Further, BPEO showed strong antioxidant activity (IC₅₀ = 7.36 μL/mL) as measured by DPPH· assay. During in situ investigation, BPEO completely inhibited AFB₁ production in model food (Phyllanthus emblica) system without altering the sensory properties and also exhibited high LD₅₀ value (14,584.54 μL/kg) on mice. In addition, BPEO was encapsulated into chitosan, characterized and tested for their potential to inhibit growth and AFB₁ production. The mean particle size, PDI and zeta potential of formed BPEO-loaded chitosan nanoparticle (CS-Np-BPEO) were performed to confirm successful encapsulation. The result revealed nanoencapsulated BPEO showed enhanced activity and completely inhibited the growth and AFB₁ production by AF-LHP-PE-4 at 0.8 μL/mL. Based on findings, it could be concluded that the BPEO and its encapsulated formulation can be recommended as a potential plant-based preservative against fungal and aflatoxin contamination of stored masticatories.

The present study was undertaken to explore the inhibitory effect of Levisticum officinale Koch. essential oil (LₒffEO) on the growth and aflatoxin B₁ secretion by Aspergillus flavus (AF-LHP-SH1, aflatoxigenic strain) causing deterioration of stored chia seeds (Salvia hispanica). The chemical profile analysis of LₒffEO by GC-MS analysis revealed the presence of α-terpinyl acetate (26.03 %) as a major component followed by terpineol <1- > (24.03 %) and citronellal (24.03 %). Results on antifungal and antiaflatoxigenic activity indicated that LₒffEO at 2.0 and 1.75 μL/mL caused complete inhibition of growth and aflatoxin B₁ production, respectively. Antifungal toxicity of LₒffEO was strongly correlated with the inhibition of ergosterol content, leakage of cellular ions, and disintegration of membrane permeability. Reduction in cellular methylglyoxal by LₒffEO indicated a novel antiaflatoxigenic mechanism of action. The LₒffEO showed moderate free radical quenching activity in DPPH assay (IC₅₀ = 26.10 μL/mL) and exhibited remarkable inhibitory efficacy against lipid peroxidation of chia seeds. In addition, LₒffEO presented strong in situ antiaflatoxigenic efficacy, and exhibited non-phytotoxic nature, acceptable sensory characteristics, and favorable safety profile (LD₅₀ = 19786.59 μL/kg), which recommends its practical utilization as a novel and safe preservative to improve the shelf life of stored chia seeds from fungal infestation and aflatoxin B₁ contamination.