Use of lichens as bioindicators of contamination by agrochemicals and metals.

The presence or absence of lichens serves as an indicator of the condition of an ecosystem and the degree to which it is contaminated by various agents, such as agrochemicals and metals. Evaluating the use of lichens as bioindicators of agrochemical contamination could provide a more comprehensive perspective of current contamination levels. Monitoring was conducted over a 4-month period in two study areas: one was a well-conserved area contaminated by metals, and the other was an area surrounded by agricultural crops contaminated by agrochemicals. Data on the presence and abundance of lichens in each study area were recorded at 10 monitoring points, a procedure that was repeated 16 times (every 15 days), and concentrations of heavy metals and "organophosphate" agrochemicals in the lichens collected were measured by means of Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-OES) and Gas Chromatography (GC), respectively. Generalized linear mixed models were used to assess abundance and richness, while general linear mixed models were used to attain Shannon diversity and Simpson dominance indices. Moreover, a multivariate analysis was performed in order to compare the lichen communities in both areas. The results indicated differences between the area contaminated by metals and that contaminated by agrochemicals in terms of abundance and Simpson's dominance index, while no differences were found in the case of the richness and diversity models. The PERMANOVA analysis additionally showed differences between the lichen communities in the two areas. The results also demonstrated that Canoparmelia caroliniana bioaccumulated metals in both areas. The levels of barium, cadmium, and sodium were higher in the area contaminated by metals, while concentrations of chromium and copper were higher in the area contaminated by agrochemicals. Finally, the concentrations of agrochemicals were higher in the area contaminated by agrochemicals and included toxic substances such as Methylparathion and Parathion, which are prohibited in Ecuador. In conclusion, this research underscores the importance of lichens as precise indicators of environmental health and contamination by agrochemicals and metals.

Determination of phytochemical contents by LC/QTOF/MS and evaluation of in-vitro biological activities of 2 Peltigera lichens from Bursa.

Lichens are symbiotic associations of algae and fungi. They are edible as food and have been used in traditional medicine for years. It is aimed to screen Peltigera praetextata (Flörke ex Sommerf.) Zopfand and Peltigera elisabethae Gyeln. phytochemically by LC/QTOF/MS and according to the constituents to evaluate the antioxidant, tyrosinase inhibitory, and antibacterial activities. In total 54 of metabolites detected by LC/QTOF/MS were common in both species. According to LC/QTOF/MS scanning results, alkaloids, iridoid glycosides, phenolics, cyanogenetic glycosides, and terpenic structures were detected. DPPH, ABTS, superoxide radical scavenging activities, and metal chelating capacity IC50 values were 84.55, 9.349; 51.27, 9.127; 95.01, 58.65 and 20.57, 70.08 µg/mL., respectively. The CUPRAC reducing power was determined as 4.69 and 9.57 TEACCUPRAC, respectively. Tyrosinase inhibitor activity were found to be 86.95 and 196.7 µg/mL. Both lichens did not show antimicrobial effects. As a result of the antioxidant and tyrosinase inhibitor activities it was seen that their activities were significant and further in vivo studies could be carried out on this lichens.

Liquid chromatography-electrospray ionization-mass spectrometry/mass spectrometry characterization of depsides and depsidones from the Chilean lichen Parmotrema perlatum.

Lichens are recognized by their unique compounds and diverse applications in food, medicines, and cosmetics. Using ultra-high pressure liquid chromatography, coupled with a high-resolution mass spectrometer, metabolomic profiling of the lichen Parmotrema perlatum, from a methanolic extract, was performed. Based on characteristic fragmentation patterns, twenty-five lichenic substances were tentatively identified including 5 depsides, 12 depsidones, 2 diphenyl ethers, 1 aromatic considered as possible artifact, 1 dibenzofuran, 1 carbohydrate, 1 organic acid, and 2 undefined compounds. To the best of our knowledge, this is a more complete report of their phytochemistry from P perlatum. Our findings of the P perlatum profile may contribute and complement the current data of the Parmotrema genus.

Contribution of Mine-Derived Airborne Particulate Matter to Ca, Fe, Mn and S Content and Distribution in the Lichen Punctelia hypoleucites Transplanted to Bajo de la Alumbrera Mine, Catamarca (Argentina).

The aim of this work was to relate the contribution of mine-derived airborne particulate matter to Ca, Fe, Mn and S content and distribution in Punctelia hypoleucites transplanted to Bajo de la Alumbrera, an important open-pit mine in Catamarca, Argentina. Lichen samples were transplanted to four monitoring sites: two sites inside the mine perimeter and two sites outside the mine. After three months, elemental distribution in samples was analyzed by microparticle-induced X-ray emission (microPIXE), and elemental concentration was determined by specific techniques: Ca and Fe by instrumental neutron activation analysis, Mn by inductively coupled plasma atomic emission spectrometry and S by a turbidimetric method. A differential distribution of S and Ca in thalli transplanted in-mine sites was detected compared to that of samples transplanted outside-mine sites. An overlap of Fe and S in the upper cortex of the apothecium section was observed, leading to infer a mineral association of both elements. Similar association was observed for Ca and S. In addition to these results, the significantly higher concentration detected for S and Mn in in-mine site samples suggests a contribution of Fe, S, Ca and Mn of mining origin to the content and distribution of these elements in P. hypoleucites. MicroPIXE complemented with Mössbauer spectroscopy analysis determined the presence of pyrite particles together with other iron-bearing minerals displaying different degrees of oxidation. These results point to a mining origin of the airborne particulate matter trapped by the lichen thalli transplanted to Bajo de la Alumbrera. These findings indicate that P. Hypoleucites acts as an excellent air quality biomonitor in the Bajo de la Alumbrera mine area.

Assessment of Cytotoxic/Antitumour Potential and in silico Study of Salazinic Acid Isolated from Parmotrema concurrens.

INTRODUCTION: Despite numerous scientific advances, cancer continues to be one of the main causes of death in the world. This situation has driven the search for promising molecules. Lichen substances have been widely described for their pharmacological potential. OBJECTIVE: The present study evaluated the antitumour potential of a depsidone isolated from Parmotrema concurrens- salazinic acid (SAL) - through in vitro, in vivo and in silico studies. METHODS: The molecule was isolated from the acetonic extract of the lichen and recrystallized in acetone. The macrophage J774, sarcoma-180 and MDA-MB-231 cell lines were used for the MTT cytotoxicity assay. The antitumor assay used a murine model (Swiss albino mice) with sarcoma-180. The animals were treated for seven consecutive days with doses of SAL (25 and 50 mg/kg) and 5-fluorouracil (20 mg/kg). RESULTS: Its purity was determined using high-performance liquid chromatography (94%), and its structure was confirmed by H1 and C13 nuclear magnetic resonance. SAL was not considered toxic to cancer cell lines, showing cell viability rates of 79.49 ± 4.15% and 86.88 ± 1.02% for sarcoma-180 and MDA-MB-231, respectively. The tumour inhibition rate was greater than 80% in the animals treated with SAL and 65% for those that received 5-fluorouracil. Simulations of molecular dynamics to estimate the flexibility of the interactions between human thymidylate synthase and derivatives of SAL and 5-fluorouracil revealed that SAL exhibited greater enzymatic interaction capacity, with highly favourable energy, compared to 5-fluorouracil. CONCLUSION: The present results demonstrate the potential of salazinic acid as a tumour inhibition agent.

Green ultrasound-assisted extraction of lichen substances from Hypotrachyna cirrhata. Ethyl lactate, a better extracting agent than methanol toxic organic solvent?

For the first time, we report a green extraction of lichen substances assisted by high power ultrasounds from Hypotrachyna cirrhata using ethyl lactate. This sustainable alternative was comparable, both in isolation and detection of lichen substances, to methanol. In the metabolomic analysis, a total of 77 lichen substances were detected comprising depsides, depsidones, dibenzofurans, organic acids, and lipids. Although the UHPLC/ESI/MS profiles were similar, the antioxidant activity was higher for the ethyl lactate extract. Ethyl lactate can replace toxic organic solvents, such as methanol, in order to provide more sustainable green chemistry methods.

A sustainable application for the extraction of lichen metabolites from Usnea cornuta: nontargeted metabolomics and antioxidant activity.

In this study, isolation and purification of lichen substances from Usnea cornuta were performed using conventional solvents, green solvents and green technologies. In addition, several lichen compounds were tentatively identified by UHPLC/ESI/MS/MS and usnic acid, diffractaic and galbinic acids were quantified as well. Limonene, ethyl lactate and methanol, were compared regarding their extraction properties and antioxidant capacities, determined by DPPH, ORAC, and FRAP assays. In the ethyl lactate, methanol and limonene extracts, 28 compounds in all, were detected for the first time by high resolution UHPLC-MS/MS fingerprinting. Untargeted metabolomics tentatively identified 14 compounds from the methanolic extract, 4 from limonene extract, and 20 metabolites from ethyl lactate extract. The green extract of ethyl lactate showed a similar antioxidant capacity to toxic methanol extract, except at ORAC assay where it was higher. Therefore, ethyl lactate can replace methanol, to provide more sustainable green chemistry methods.

Beneficial Effects of Liposomal Formulations of Lichen Substances: A Review.

Lichens are commonly used as essential traditional medicines to treat various conditions, including skin disorders, wounds, digestive, respiratory, obstetric, and gynecological problems in many cultures in Africa, Asia, Europe, Haitian, Oceania, and North and South America. Lichens have been deeply investigated for their phytochemical properties and, to date, numerous compounds (also known as substances) have been successfully isolated from the extracts. However, the low solubility and bioavailability of pure lichen substances have been widely recognized as significant issues hindering their biological applications. Recently, several groups have investigated the properties and the potential applications of lichen metabolites-based liposomal formulations and revealed a substantial improvement in their solubility, bioactivity, and toxicity in the animal. Thus, in this topical review, we aimed to provide an overview of liposomal structures, the efficacy of liposomal formulations, as well as their beneficial effects as compared to the free compounds themselves.

Physiological Response and Multi-Elemental Content in Lichens Growing on Agricultural Fences: A Pilot Study Comparing No-Tillage and Organic Cropping.

The objective of this study is to compare the physiological response (content and degradation of photosynthetic pigments, membrane oxidation products and soluble proteins) and multi-element content of Ramalina celastri (lichenized fungi) growing on agricultural fences with no-tillage (associated with transgenic crops and agrochemical application), organic cropping and a non-cultivated area. We found that R. celastri did not differ in its physiological response to agricultural practices, except for the contents of chlorophyll b and phaeophytin a which were high in both cultivated areas. Lichens growing in organic cropping fields have higher arsenic, chromium, uranium and internal transition elements common in the earth's crust, possibly due to the greater resuspension of the material during soil tillage. Lichens that grow on posts close to no-tillage field had higher bromine contents (present in numerous pesticides). We found evidence that R. celastri behaves as a tolerant species to air pollution in agricultural environments.

Continental and Antarctic Lichens: isolation, identification and molecular modeling of the depside tenuiorin from the Antarctic lichen Umbilicaria antarctica as tau protein inhibitor.

Alzheimer´s disease (AD) is the most common form of dementia involving Aß and tau protein. So far, AD cure remains elusive, but considering that AD progresses throughout tau pathology, which turns tau protein an appropriate target, besides tau is also included in other neurodegenerative disorders named as tauopathies. Here, we have isolated seventeen compounds belonging to six lichens species. Due to scarce of spectroscopic data of the compound 5,7-dihydroxy-6-methylphthalide, we explained their structural elucidation based on NMR data. In this study, we show that only tenuiorin from Umbilicaria antarctica inhibited 50% of tau 4R at 100 µM. Then, we shown that molecular interactions of tenuiorin with the steric zipper model of the hexapeptide 306VQIVYK311 were studied by docking calculations and the results suggested that tenuiorin forms both hydrogen bonds with lysine and glutamine side chains and forms several hydrophobic interactions with valine and lysine from 306VQIVYK311 motif.

Epanorin, a lichen secondary metabolite, inhibits proliferation of MCF-7 breast cancer cells.

BACKGROUND: Epanorin (EP) is a secondary metabolite of the Acarospora lichenic species. EP has been found in lichenic extracts with antimicrobial activity, and UV-absorption properties have been described for closely related molecules; however, its antiproliferative activity in cancer cells has not yet been explored. It has been hypothesized that EP inhibits cancer cell growth. MCF-7 breast cancer cells, normal fibroblasts, and the non-transformed HEK-293 cell line were exposed to increasing concentrations of EP, and proliferation was assessed by the sulforhodamine-B assay. RESULTS: MCF-7 cells exposed to EP were examined for cell cycle progression using flow cytometry, and DNA fragmentation was examined using the TUNEL assay. In addition, EP's mutagenic activity was assessed using the Salmonella typhimurium reverse mutation assay. The data showed that EP inhibits proliferation of MCF-7 cells, and it induces cell cycle arrest in G0/G1 through a DNA fragmentation-independent mechanism. Furthermore, EP's lack of overt cytotoxicity in the normal cell line HEK-293 and human fibroblasts in cell culture is supported by the absence of mutagenic activity of EP. CONCLUSION: EP emerges as a suitable molecule for further studies as a potential antineoplastic agent.

Toxicological effects of Ramalina aspera (lichen) on Biomphalaria glabrata snails and Schistosoma mansoni cercariae.

In this study, the molluscicidal activities against Biomphalaria glabrata and cercaricidal activities against Schistosoma mansoni of the ether extract of Ramalina aspera were evaluated. Additionally, toxicity parameters were evaluated at sublethal doses in terms of the influence of the extract on the fertility and fecundity of snails, as well as morphological alterations and quantification of their immunological cells. A test with Artemia salina was also carried out, in order to verify the environmental toxicity of the compound. The ether extract of R. aspera, in which divaricatic acid was identified as the major compound, demonstrated molluscicidal activity at low concentrations against both embryos (LC90 of 22.78, 24.23, 16.63 and 16.03 µg mL-1 for the gastrula, blastula, trochophore and veliger, respectively) and against adult snails (LC90 of 8.66 µg mL-1), after 24 h of exposure. At the sublethal doses, it was possible to observe a decrease in fecundity and quantitative and morphological changes in the defense cells of the exposed snails. In addition, the extract of R. aspera showed a cercaricidal effect on S. mansoni from the concentration of 5.0 µg mL-1, while showing low toxicity to Artemia salina. The ether extract of R. aspera demonstrated effective molluscicidal activity on embryos and adult snails of the species B. glabrata, cercariae of S. mansoni, and presenting low toxicity on Artemia salina. In this way, it could be considered a promising compound in the development of future molluscicidal and cercaricidal agents, thus helping to combat schistosomiasis.

On the synthesis of vitamin d in the darkness / Sobre la síntesis de vitamina D en la oscuridad

Published evidence reports the existence of two routes for the transformation of 7-dehydrocholecalciferol into previtamin D3: a photochemical route with the participation of UVB photons and another route that occurs in the darkness. Several reports appears to support the presence of these two routes in some mammals, birds, nonvascular plants (e.g.: mosses), vascular plants (e.g.: angiosperms) and lichens. The reviewed evidence suggests that in the darkness, the synthesis of vitamin D3 follows the same scheme of the photochemical pathway, but at a reduced rate respect to the synthesis under UVB radiation. The process of vitamin D synthesis in the dark, then, may be taken as an insurance for survival, at least for mammals and birds. The low rate of the synthesis of vitamin D3 in the absence of light produce low concentrations of vitamin D3 metabolites in plasma. Long term survival under these circumstances might be possible through upregulation of vitamin D receptors (VDRs). In mole rats (South African rodents that live in the dark in underground tunnels), the reduced rate of vitamin D3 synthesis produce low levels of plasma vitamin D3 and their metabolites 25(OH) D3 and 1α,25OH2 D3 . The fact that Kd and ßmax of the complex 1α,25(OH)2 D3 -VDR from the intestinal mucosa, kidneys and the Harderian glands of the mole rat Heterocephalus glaber are significantly different in each one of these tissues, is interpreted as an indicator that the VDRs are, in each tissue, adapted to the maintenance of normal physiological functions. (AU)

Varios trabajos publicados han informado que existen dos mecanismos para la transformación de 7-dehidrocolecalciferol en previtamina D3 : uno iniciado con el auxilio de fotones UVB y un segundo que ocurre en la oscuridad, sin el auxilio de radiación ultravioleta. Una serie de publicaciones contienen información que apoya la presencia de estos dos mecanismos en mamíferos, pájaros, plantas no vasculares (musgos), vasculares (angiospermas) y líquenes. La evidencia revisada sugiere que, en la oscuridad, la síntesis de vitamina D3 sigue el mismo esquema que la ruta fotoquímica y generalmente ocurre a una tasa reducida respecto de la síntesis bajo irradiación ultravioleta. La operación de la síntesis de vitamina D3 en la oscuridad, por lo menos para mamíferos y pájaros, puede tomarse como un reaseguro de sobrevida. La reducida tasa de síntesis de vitamina D en ausencia de luz produce bajas concentraciones en plasma de los metabolitos de la vitamina. La sobrevida saludable en estas condiciones sería posible mediante upregulation de los receptores. En las ratas topo Heterocephalus glaber (roedores sudafricanos que viven en permanente oscuridad, en túneles subterráneos), la reducida tasa de síntesis de vitamina D3 es la causa de los bajos niveles plasmáticos de la vitamina y sus metabolitos: 25(OH)D3 y 1α,25OH2 D3 . El hecho de que el Kd y ßmax del complejo 1α,25(OH)2 D3 -Vitamina-D-Receptor del intestino, riñón y glándulas de Harder de Heterocephalus glaber sean significativamente diferentes entre sí indicaría que los receptores se han modificado para mantener funciones fisiológicas normales en cada tejido. (AU)

Epanorin, a lichen secondary metabolite, inhibits proliferation of MCF-7 breast cancer cells

BACKGROUND: Epanorin (EP) is a secondary metabolite of the Acarospora lichenic species. EP has been found in lichenic extracts with antimicrobial activity, and UV-absorption properties have been described for closely related molecules; however, its antiproliferative activity in cancer cells has not yet been explored. It has been hypothesized that EP inhibits cancer cell growth. MCF-7 breast cancer cells, normal fibroblasts, and the non-transformed HEK-293 cell line were exposed to increasing concentrations of EP, and proliferation was assessed by the sulforhodamine-B assay. RESULTS: MCF-7 cells exposed to EP were examined for cell cycle progression using flow cytometry, and DNA fragmentation was examined using the TUNEL assay. In addition, EP's mutagenic activity was assessed using the Salmonella typhimurium reverse mutation assay. The data showed that EP inhibits proliferation of MCF-7 cells, and it induces cell cycle arrest in G0/G1 through a DNA fragmentation-independent mechanism. Furthermore, EP's lack of overt cytotoxicity in the normal cell line HEK-293 and human fibroblasts in cell culture is supported by the absence of mutagenic activity of EP. CONCLUSION: EP emerges as a suitable molecule for further studies as a potential antineoplastic agent.

Morphophysiological variation and metal concentration in the thallus of Parmotrema tinctorum (Despr. ex Nyl.) Hale between urban and forest areas in the subtropical region of Brazil.

Anthropic activities such as the emission of pollutants resulting from industrial and agropastoral activities promote several changes in urban and forest areas. Lichens are organisms that are used in air quality evaluations due to their sensitivity to these changes. The aim of this study is to analyze the presence of morphophysiological damages and the metal concentration in samples of the lichen Parmotrema tinctorum, in urban and forest areas, checking for possible parameter variations between these areas, in the different matrices and seasons in the Southern region of Brazil. Six areas were selected (urban and forest), assigned to the rural/urban and urban/industrial matrices belonging to the watershed of the Rio dos Sinos, Brazil. The following parameters were analyzed: Index of photobiont vitality (IPV), photosynthetic pigments (chlorophyll, carotene, and pheophytin) and metals (Cu, Cr, Zn, Pb, and Ni) in the thallus of the lichen. The data were analyzed by an ANOVA one way, Pearson correlation test, and principal component analysis (PCA). Variations in the morphophysiological parameters were recorded in all the areas revealing significant differences. The lowest IPV values and highest concentration of metals were recorded in the urban environment, while low chlorophyll levels were found in the forest areas. The PCA showed a distinction between the areas and the season. The level of urbanization, vehicle traffic, and the weather conditions might have influenced the results. The use of P. tinctorum, the index of photobiont vitality and chlorophyll and pheophytin content, has proved to be an efficient tool to diagnose the air quality in the areas analyzed, allowing its use as a model in air monitoring studies, both in urban and forest areas, as well as in distinct matrices in the subtropical region.

Tracking atmospheric dispersion of metals in Rio de Janeiro Metropolitan region (Brazil) with epiphytes as bioindicators.

The atmospheric dispersion atmospheric plume of the metropolitan region and neighborhoods of the city of Rio de Janeiro was investigated through elemental analyzes (Na, K, Al, Mn, Pb, Ni, Cr) and stable lead isotopes in two epiphytes; the lichen Parmotrema crinitum (Ach.) and the bromeliad Tillandsia usneoides (L.). All the elements had lower concentrations in epiphytes than in rocks of the local geological basement, with the exception of K, which was similar to rocks. This behavior was attributed to the nutritional essentiality and abundance of this element in the environment. The concentrations of Na, Pb and the 206Pb/207Pb and 208Pb/206Pb ratios indicated the presence of spatial gradients in the path (a) sea shore → continent, and (b) urban center → farmlands. These patterns were associated with the mesoscale atmospheric circulation regime, in which, during the arrival of cold fronts, air masses from the metropolitan region of Rio de Janeiro are transported towards the mountainous region of Teresópolis, which acts as a barrier to the dispersion of pollutants. The bromeliad Tillandsia usneoides and the lichen Parmotrema crinitum were linked to Na and Pb atmospheric levels, respectively.

Characterization of nitrogen deposition in a megalopolis by means of atmospheric biomonitors.

An increase of nitrogen deposition resulting from human activities is not only a major threat for global biodiversity, but also for human health, especially in highly populated regions. It is thus important and in some instances legally mandated to monitor reactive nitrogen species in the atmosphere. The utilization of widely distributed biological species suitable for biomonitoring may be a good alternative. We assessed the suitability of an ensemble of atmospheric biomonitors of nitrogen deposition by means of an extensive sampling of a lichen, two mosses, and a bromeliad throughout the Valley of Mexico, whose population reaches 30 million, and subsequent measurements of nitrogen metabolism parameters. In all cases we found significant responses of nitrogen content, C:N ratio and the δ15N to season and site. In turn, the δ15N for the mosses responded linearly to the wet deposition. Also, the nitrogen content (R2 = 0.7), the C:N ratio (R2 = 0.6), and δ15N (R2 = 0.5) for the bromeliad had a linear response to NOx. However, the bromeliad was not found in sites with NOx concentrations exceeding 80 ppb, apparently of as a consequence of excess nitrogen. These biomonitors can be utilized in tandem to determine the status of atmospheric nitrogenous pollution in regions without monitoring networks for avoiding health problems for ecosystems and humans.

Trace element biomonitoring in the Peruvian andes metropolitan region using Flavoparmelia caperata lichen.

In the present study, in situ lichens (Flavoparmelia caperata) were used to assess the deposition of atmospheric trace elements in the metropolitan area of Huancayo (Junín, Peru). In total, ten sampling sites were chosen and categorized as urban, peri-urban (rural-urban) and rural areas according to land use. In addition, samples were also collected from a non-contaminated area categorized as a control site. The concentrations of 16 trace elements were measured using an inductively coupled plasma mass spectrometer (ICP-MS) and examined by enrichment factor (EF), hierarchical cluster analysis (HCA), and principal component analysis (PCA). Twelve of the 16 trace elements in urban and peri-urban sites present concentration higher than those at the rural and control sites (p < 0.05). The EF results revealed significant enrichment (at least twice that of the control site) of Ba, Cr, Cd, Pb, Sb, V, and Zn at most sites. PCA and HCA showed that more elements were derived from vehicular sources and fewer from agricultural and natural sources.

Evaluación de la actividad antifúngica del extracto de Usnea laevis en hongos fitopatógenos / Evaluation of the antifungal activity of the lichenic extract of Usnea laevis in phytopathogenic fungi

Los líquenes son producto de una asociación simbiótica entre un hongo y alga y/o cianobacteria; esta simbiosis produce una serie de metabolitos secundarios o sustancias liquénicas únicas, las mismas que son aisladas a partir de sus extractos y que han presentado una marcada actividad antibiótica y antifúngica. Considerando que en el Ecuador no se tienen antecedentes acerca de este tipo de estudios, el objetivo planteado fue el evaluar la actividad antifúngica del extracto liquénico de Usnea laevis frente a hongos fitopatógenos. En el presente estudio las sustancias liquénicas almacenadas en el talo de Usnea laevis fueron extraídas con metanol y etanol. La actividad antifúngica in vitro de estos extractos fue probado frente a géneros de hongos fitopatógenos previamente aislados (Aspergillus, Penicillium y Rhizopus stolonifer); añadiéndolo al medio de cultivo Agar Papa Dextrosa (PDA) a una concentración de 0,5%; determinándose el porcentaje de inhibición. Los datos fueron analizados estadísticamente y se realizó la clasificación del extracto en base a su porcentaje de inhibición según la OILB. El extracto metanólico y etanólico de Usnea laevis mostró una mayor efectividad frente a Rhizopus stolonifer, inhibiendo más del 50% de su crecimiento, a diferencia de Penicillium y Aspergillus cuyo porcentaje de inhibición fue mucho menor y mostraron diferencias estadísticamente significativas. El extracto liquénico fue moderadamente tóxico para Rhizopus stolonifer, ligeramente tóxico para Aspergillus sp 1 y Penicillium sp e inocuo para Aspergillus sp 2.

A lichen is an organism product of a symbiotic association between a fungus and algae and / or cyanobacteria; this symbiosis produces many secondary metabolites or unique lichen substances, which are isolated from their extracts and show a marked antibiotic and antifungal activity. Considering there is no background on this type of studies in Ecuador, the aim of this study was to evaluate the antifungal in vitro activity of Usnea laevis extract on phytopathogenic fungi. The lichenic substances stored in the thallus of Usnea laevis were extracted with methanol and ethanol. The in vitro antifungal activity of these extracts was tested against phytopathogenic fungal genera previously isolated (Aspergillus, Penicillium and Rhizopus stolonifer) . They were adding to Potato Dextrose Agar (PDA) culture medium at a concentration of 0.5%, and the inhibition percentage was determined. Data were statistically analyzed and the extract was classified based on its inhibition percentage according to the OILB. Methanolic and ethanolic extracts of Usnea laevis showed greater effectiveness against Rhizopus stolonifer, inhibiting more than 50% of its growth, unlike Penicillium and Aspergillus, whose inhibition percentage was lower and showed significant differences. The liquid extract was also moderately toxic to Rhizopus stolonifer, slightly toxic to Aspergillus sp 1 and Penicillium sp and harmless to Aspergillus sp 2.

Toxicity of Usnic Acid from Cladonia substellata (Lichen) to embryos and adults of Biomphalaria glabrata.

This study reports the molluscicidal activity of usnic acid isolated from Cladonia substellata Vanio (lichen) on embryos at various stages of development and in adult mollusks of Biomphalaria glabrata. The toxicity of usnic acid was also evaluated through Artemia salina larvae mortality. Usnic acid was extracted with diethyl ether, isolated, purified, and its structure confirmed by analyzing the spectra of proton nuclear magnetic resonance. LC90 for 24 h of exposure were 1.62, 4.45, 5.36, and 4.49 µg mL-1 for blastula, gastrula, trocophore, and veliger embryonic stages, respectively, and 3.45 µg mL-1 for adult snails; LC50 of usnic acid against A. salina was 2.46 µg mL-1. LC90 assessed 7 days after exposure was 2.56 µg mL-1 for adult mollusks. In conclusion, these findings demonstrate that under laboratory conditions usnic acid has teratogenic and molluscicide potential to control the aquatic snail B. glabrata and may prove to be a promising candidate in the search for new molluscicide agents, but further detailed studies on its molluscicidal effect and possible environmental effects are needed.