Exploration of photoprotective and antibiotic activity of 20 wild Polypodiaceae ferns from Costa Rica

Exploration of photoprotective and antibiotic activity of 20 wild Polypodiaceae ferns from Costa Rica | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Exploration of photoprotective and antibiotic activity of 20 wild Polypodiaceae ferns from Costa Rica Yaclyn Salazar-Chacon, Maria José Gutierrez-Bolaños, Jimena Padilla-Cordero, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2533922/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 Jan, 2024 Read the published version in Scientific Reports → Version 1 posted 10 You are reading this latest preprint version Abstract Skin disorders affect millions of people all over the world. There are limited options to treat dermal illnesses such as vitiligo, psoriasis, and atopic dermatitis (eczema). Central American ferns are a potential source of bioactive metabolites against those diseases. Currently, Polypodium leucotomos Poir. (Phlebodium aureum (L.) J. Sm. synonym) is the only one being commercially utilized for this purpose. In this work, we evaluated the concentration of the skin bioactive compounds: quinic and chlorogenic acid, in the extract of 20 wild ferns from Costa Rica. We also evaluated the antimicrobial capabilities of the raw extracts of wild ferns and the sun protection factor (SPF) of the extracts. We found 19 out of 20 have either an important concentration of the compounds mentioned above or antimicrobial properties. Also, most samples result in higher SPF than P. aureum’s rhizome. We also have studied the fern acclimatization, at different shading conditions, finding a significant influence of the culturing conditions on metabolite production. After acclimatization. So far, we demonstrate that various ferns included in this study are a potential source of treatments for skin conditions. Physical sciences/Chemistry/Medicinal chemistry Physical sciences/Chemistry/Chemical biology/Natural products Biological sciences/Plant sciences quinic acid vitiligo fern chlorogenic acid skin antibiotic bacteria Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1. Introduction Ferns are primitive non-flowering vascular plants, and many of them have shown medicinal properties. Several ferns have been studied as bio-herbicides, hepatoprotectives, cytotoxic, antihyperglycemic, trypanocidal, antimicrobial, antinociceptive, and immunomodulatory 1 . Nutritional, nutraceutical and pharmacological utilization are still limited at a commercial scale, with some exceptions, such as kalawalla ( Phlebodium aureum formerly known as Polypodium leucotomos ). The extract of P. aureum has been used to treat skin diseases since the ‘80s, and there are many commercial preparations based on the hydrosoluble extract of their rhizomes, such as Fernblock®, Heliocare®, Difur®, and others. P. aureum extracts have been commercialized mainly in Europe despite this plant is endemic to Central America 2 . Oral or topic formulations containing P. aureum extract are recognized for their photoprotectant bioactivity and their strong antioxidant properties, which have been related to their naturally occurring phenolic compounds such as p -coumaric, ferulic, caffeic, vanillic, and chlorogenic acid 3 . Commercial aqueous extract of the leaves of P. aureum ( marketed as Fernblock®, Fernplus®, Fernmed®, and Fernage®) contain a standardized 0.6–1.3% total phenolic compounds concentration and 0.4–0.9% quinic acid concentration 4 . Mechanisms of action of these commercial products involve free radical scavenging and regulation of several genes, such as metalloproteinases 5 , tyrosinase inhibition 6 , and others. Also, natural phenolic compounds are known for absorbing UV radiation, similar to synthetic compounds, such as homosalate or dibenzalacetone do so. Then fern extracts are an excellent alternative for bio-based wide-spectrum sun blockers and sunscreen 7 . Also, fern extracts have been found active to prevent UV damage during clinical trials 8 , preventing sunburns, and subsequently skin cancer. Fern-derived nutraceutical products are an alternative for some skin disorders with unavailable treatment options, such as eczema or vitiligo. Also, skin microbiome imbalance and traditional antibiotic utilization contribute to developing and speeding up skin depigmentation 9 . For example, Staphylococcus spp. is more abundant on the skin of patients with vitiligo 10 , 11 . Some aqueous fern rhizome extracts showed antibacterial properties against gram-negative bacteria (such as Escherichia coli ) and gram-positive (such as S. aureus ) 12 . Other Central American Polypodiaceae and Pteridophyta ferns have similar metabolic pathways to P. aureum 13 , and also have been used for the same applications in traditional medicine 14 . Although some wild ferns are not feasible to reproduce in a greenhouse, there is an enormous potential for the discovery and commercialization of medicinal ferns in Central America 15 . Also, growing conditions such as climate, altitude, light incidence, and substrate composition can be used to improve or standardize metabolite production 16 , 17 . Therefore, we hypothesize that other ferns can have similar or superior activity compared to P. aureum , in terms of UV protection and metabolite concentration. Some of them can be potentially more active or more economic to produce than kalawalla is. In this work, we explored wild and acclimatized ferns in terms of amounts of quinic acid, chlorogenic acid, and total polyphenolic compounds, as potential alternatives to treat skin disorders. 2. Materials And Methods 2.1 Microorganism strains The strains utilized in this study were Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis ATCC 12228, Pseudomonas aeruginosa ATCC 9027, and Escherichia coli ATCC 25922. 2.2 Fern samples Twenty wild native fern species were collected from the forest at 13 locations. Figure 1 (A) summarizes the identified species and their location. Both rhizomes and leaves were collected for each plant, oven-dried for 72 h at 40°C, and ground to 1 mm. 2.3 Quinic and chlorogenic acid determination 0.5 g dry material (rhizome or leaves) was extracted in 1.5 mL 75% methanol. Afterwards, it was filtered through a 0.2 µm membrane. The extract was dried into an analytic evaporator ZipVap Zanntek (from Glas-Col®, Terre Haute, IN) at 40°C under nitrogen. Then, it was reconstituted in 500 µL 75% methanol. The reconstituted extract was analyzed by HPLC, using a solvent gradient described in Table 1 . The column utilized is an Acclaim 120 (C-18 (4,6 ×150) mm, 5 µm) (from Thermo Scientific Dionex, CA) at 35°C. A diode array detector was used to detect quinic acid (QA) and chlorogenic acid (CGA) at 215 and 325 nm, respectively, against 0-2000 mg/L QA and 0-600 mg/L CGA standard curves. Table 1 Starting, interception and ending points for linear gradients of mobile phase, utilized to quantify quinic and chlorogenic acid through HPLC. Point Time (min) Flow rate Acetonitrile Metanol 0.1%TFA Starting 0 0.8 10% 0% 90% Intersection 5 0.8 10% 10% 80% Intersection 8 0.8 10% 20% 70% Intersection 10 0.8 10% 40% 50% Intersection 13.5 1.5 10% 10% 80% Ending 15 0.8 10% 0% 90% 2.4 Total polyphenolic content (TPC) quantification 0.1 g of each sample was mixed with 3 mL of 75% methanol and sonicated for 10 min. Then, the mixture was centrifuged for 5 min, and the methanolic extract was collected. The remaining solid was extracted two additional times following the same procedure. All the extracts were combined and diluted to complete 10 mL (total volume). Then, Folin-Ciocalteu’s colorimetric method was used to determine total phenolic content, as described in our previous work 18 , using gallic acid as standard. Each sample has three replicates. Both S. dissimile specimens were combined in equal amounts for this analysis. 2.5 Antibiotic activity test In two replicates, we followed the agar well diffusion method 19 , a modification of the Kirby-Bauer susceptibility protocol 20 . These experiments were conducted on Mueller Hinton agar plates. 100 µL of each microorganism at 0,5 of McFarlan scale (1,5x 10 8 UFC/mL) in NaCl 0.85% were plated into a sterile 90x15mm petri dish containing 25 mL of Mueller Hinton agar. Then, four 6 mm wells were made in the agar at the same distance, three wells were filled with 50µL of 0.1g/mL fern extract, and the fourth well was filled with 50 µL of 50%v/v acetone (as negative control). Additionally, for the positive control, the same procedure was carried out, but 3 well were filled with 50µL of 1000 µg/mL of Penicillin/Streptomycin (from GIBCO, NY), and the fourth well was filled with 50 µL of 50%v/v acetone (as negative control). Plates were incubated for 24 h at 35°C, and inhibition rings were measured. Two replicates were used for each condition. Microbial activity was calculated according to (1). $$\text{%}RPI=\frac{({D}_{sample}-{D}_{negative control})}{\left({D}_{positive control}\right)}x100$$ 1 % RPI is the Relative Percentage of Inhibition, and D sample , D negative control , and D positive control are the diameters of the inhibition zones indicated in the subscripts. 2.6 Solar protection factor (SPF) determination The same extracts prepared for TPC analysis were diluted 100 times, and the UV-Vis spectra were recorded for each sample repetition between 190–800 nm in a spectrophotometer Evolution 350 (from Thermo Scientific, Massachusetts, USA). Data were exported and analyzed according to Mansur Eq. 2 1 . SPFs were adjusted to the corresponding concentration found in the dry material (leaves or rhizome), and the average of 3 commercial sunscreens (SPF 50) was used as reference: Nivea Sun®, Eucerin®, and Vicky face® sunscreens. 3 repetitions had been used. 2.7 Acclimatization of selected ferns Six fern species were selected to grow under controlled conditions to study the acclimatization effect over metabolite production. Campyloneurum latum T. Moore, Niphidium nidulare (Rosenst.) Lellinger, Phlebodium pseudoaureum (Cav.) Lellinger, Serpocaulon attenuatum (C. Presl) A.R. Sm., S. sessilifolium (Desv.) A.R. Sm. y S. triseriale (Sw.) A.R. Sm. were collected from the wild at the locations mentioned in Fig. 1 (B). The samples were reproduced at Macho River Biological Station, located at 9°46'00" N, 83°46'00" W, a 1600 mamsl, in Orosí, Cartago, Costa Rica. The reported temperature range at the location is 18,3–26,5°C (average 22,3°C), and the annual precipitation is 3120 mm. 68 plants of each specie were distributed under 50% and 70% shade (covered with saran fabric) in 1–3 L plant pots containing a substrate composed by: peat (1/4), organic fertilizer (1/4), grounded coconut fiber (1/4) and coconut shell chunks (1/4). They were harvested after a year, processed, and analyzed using the same methods described for wild ferns. 3. Results And Discussion 3.1 Content of quinic and chlorogenic acids of wild ferns Figure 2 summarizes the metabolite content of leaves and rhizome of the 20 wild ferns included in this study. Polypodium leucotomos is a synonym for Phlebodium pseudoaureum (sensu stricto) or Phlebodium aureum (sensu lato). Quinic acid was present in eleven leaf extracts and eight rhizome extracts. Notwithstanding the commercial product elaborated from the rhizome extract, we decided to include the leaves, considering that metabolites could be distributed all over the ferns. Interestingly, quinic acid was detected in leaves of Pecluma consimilis (Mett.) M.G. Price, Polypodium colpodes Kunze, P. plesiosorum Kunze and Serpocaulon dissimile (L.) A.R. Sm., but it was not detected in the rhizomes of those species. Also, quinic acid was detected in both rhizomes and leaves of Niphidium crassifolium (L.) Lellinger, Pleopeltis macrocarpa (Bory ex Willd.) Kaulf., P. montingena (Maxon) A.R. Sm. & Tejero, P. myriolepis (Christ) A.R. Sm. & Tejero, Serpocaulon sessilifolium (Desv.) A.R. Sm. and S. triseriale (Sw.) A.R. Sm., but the concentration of leaves was higher in most of them, except for N. crassifolium and S. sessilifolium . Quinic acid was not detected in P. aureum leaves, but it accounts for 11.2 mg/g in the rhizome. Figure 2 (A) contains the quinic acid concentration found on fern leaves. Three species showed a quinic acid content 56–67% lower, respecting our reference value ( P. aureum rhizome): P. consimilis , P. myriolepis , and P. plesiosorum . Another five species have ranged from similar levels to 30% higher quinic acid content (than P. aureum rhizome): N. crassifolium , P. montigena , P. colpodes , S. dissimile , and S. sessilifolium . Finally, the leaves of three fern species contain between 1.7 to 2.7 fold-up respecting P. aureum rhizome: P. macrocarpa , S. dissimile , and S. triseriale . Figure 2 (C) shows quinic acid in rhizomes. Four species ( P. macrocarpa , P. montigena P. myriolepis , and S. triseriale ) contain lower quinic acid than P. aureum . But also, three species contain 1.5–2.3 higher quinic acid than P. aureum : N. crassifolium , N. nidulare , and S. sessilifolium. According to the literature, an analysis of Fernblock®, the commercial extract from P. aureum accounts for 70 g/L quinic acid. The three species with higher quinic acid concentration are potentially a more economical source of this metabolite 4 . Also, phenolic compounds account for 1% of commercial Fernblock® 4 , composed mainly of 3,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, vanillic acid, caffeic acid, p- coumaric acid, ferulic acid, 4-hydroxycinnamoyl-quinic acid, and five isomers of chlorogenic acid 22 . Chlorogenic acid is a dietary polyphenol 23 , linked with activity against inflammation, skin diseases, and others 23 , 24 , so we had included its quantification in this study. Chlorogenic acid was present in leaves, but not in rhizomes of P. pseudoaureum , P. macrocarpa, P colpodes, P. plesiororum , P. ursipes Moritz ex C. Chr. and S. triseriale . Four species C. amphostenon (Kunze ex Klotzsch) Fée, C. latum, P. myriolepis , and Polypodium sp , contain chlorogenic acid in both leaves and rhizomes. Figure 2 (D) shows the chlorogenic acid content of wild ferns included in this study. Although chlorogenic acid is reported as a phenolic component of P. aureum commercial formulations 22 , we did not find a detectable concentration on the wild rhizome, because of its very low concentration (if present). However, five rhizome extracts contain chlorogenic acid: C. amphostenon, C. latum , N. nidulare , P. myriolepis , and Polypodium sp. , ranging from 0.8–4.7 mg/g DM. Figure 2 (B) show chlorogenic acid content in leaves. P. aureum ’s leaves contain around 3.7 mg/g chlorogenic acid, and another seven ferns contain around 0.8–3.7 mg/g of this metabolite ( S. triseriale, Polypodium sp., P. plesiosorum, P. colpodes, P. macrocarpa, C. latum , and C. amphostenon ). Two fern leaves contain significantly higher chlorogenic acid content (7.9–8.5 g/L): P. ursipes and P. myriolepsis . From all these results, we highlight the high quinic acid content in N. nidulare and S. sessilifolium rhizomes, and P. macrocapa leaves. Other elements to consider in the future, to determine the feasibility of industrialization of these wild ferns, are reproduction, culturing, and toxicity. P. leucotomos extract is safe for human consumption. Some other ferns are known to contain ptaquiloside (a nor-sesquiterpene), and sesquiterpenes exhibiting mutagenic, teratogenic, clastogenic, and carcinogenic activities 25 . 3.2 Total Polyphenolic Content of wild ferns Figure 3 summarizes the TPC of the leaves and rhizomes of the ferns. TPC in leaves (Fig. 3 A) is found the highest for P. consimilis (74.2 mgGAE/gDM). Two other fern leaves contain TPC considerably higher than P. aureum leaves (41.2 mgGAE/gDM): S. dissimile (55.3 mgGAE/gDM) and P. divaricata (56.8 mgGAE/gDM). Other five fern species contain similar TPC (± 6.6 mg mgGAE/gDM) than P. aureum leaves: P. montigena (43.9 mgGAE/gDM), P. macrocarpa (42.64 mgGAE/gDM), S. sessilifolium (38.84 mgGAE/gDM), P. colpodes (35.8 mgGAE/gDM), and Polypodium sp. (34.6 mgGAE/gDM). Interestingly, most of both leaves’ samples (Fig. 3 A) and rhizome’s samples (Fig. 3 B) contain higher TPC than P. aureum rhizome (commercially used), except N. nidulare’s and C. latum’s leaves; and, C. amphostenon ’s, P. colpodes ’s and C. latum ’s rhizomes despite the last two are virtually the same TPC. In rhizomes, seven species have shown the highest TPC particularly high: N. nidulare (84.10 mgGAE/gDM), P. divaricata (71.73 mgGAE/gDM), P. ursipes (68.17 mgGAE/gDM), N. crassifolium (70.00 mgGAE/gDM), P. myriolepis (56.87 mgGAE/gDM), P. consimilis (54.58 mgGAE/gDM), and P. montigena (40.00 mgGAE/gDM). 3.3 Antibacterial properties of fern extracts Figure 4 summarizes the relative percent of inhibition of the extracts. The leaves of N. crassifolium, P. aureum, P. macrocarpa, P. montigena, P. myriolepis, P. colpodes, S. dissimile , and S. triseriale were found active against both S. aureus and S. epidermis , showing similar relative inhibition to the positive control (around 40–62%). All the other extracts from leaves do not possess antimicrobial activity. The activity of rhizome extracts can be divided into three groups. The strongest relative inhibition against S. aureus (Fig. 4 C) was found in the range of 65–80% for the extracts of N. crassifolium , P. montigena , and S. loriceum . The first two extracts show a similar inhibition against S. epidermis (Fig. 4 D) too. An intermediate range of the strength of relative inhibition (45–65%) was observed for N. nidulare , P. macrocarpa , and S. triseriale , against both bacteria ( S. aureus and S. epidermis) . Extracts from P. montigena also showed an intermediate inhibition against S. epidermis . Finally, there is a group with a moderate relative inhibition (< 45%), including the rhizomic extracts from C. xalapense Fée, P. consimilis , and S. fraxinifolium , with antimicrobial against S. aureum , and the extracts from P. ursipes against both S. aureum and S. epidermis . Quinic and chlorogenic acids are reported to be antibacterial against several strains of S. aureus , E. coli , some bacillus , and others 26 , 27 . However, in our results, none of both compounds showed any activity against the strains evaluated here under the same conditions tested for the fern extracts (data not shown). Yet, some other compounds such as flavonoids (flavones, flavonols, flavanones), and organic acids (aliphatic and aromatic acids) are known for being antimicrobial 28 . Therefore, we evaluated the crude extracts of the wild ferns against the gram-negative: P. aeruginosa and E. coli , and the gram-positive S. aureus and S. epidermidis . Our results show that the ferns possess antimicrobial against the gram-positive bacteria tested but not against the gram-negative ones. (Fig. 4 ). Remarkably, most skin infections involve gram-positive bacteria, and antibiotic resistance is occurring more frequently in them 29 . 3.4 Solar Protection Factor (SPF) of fern extracts Figures 5 A and 5 B is showing the SPF for 19 fern species included in this study. Interestingly, almost all the samples have shown higher SPF than P. aureum rhizomes, except just N. nidulare ’s leaves. The highest SPF in leaves (Fig. 5 A) is P. consimiles (29), S. sessilifolium (28), and S. dissimile (28). Also, highest SPF in rhizomes is P. divaricata (18), P. ursipes (15), and P. consimiles (13). We did not find a correlationship between TPC, quinic acid, or chlorogenic acid concentration and SPF. UV absorption is probably influenced by more than one compound, and according to Mensul’s equation, compounds with high absorptivity at 300–305 nm are the most important contributors to SPF. It is very likely, most compounds responsible for UV absorption are polyphenols, although, some of them could have maximum absorption at other wavelengths or low molar absorptivities, and this is why there is no correlationship between SPF and TPC. Selected ultraviolet spectra containing P. aureum and the samples with the highest SPF are shown in Figs. 5 C and 5 D. All the samples cover the whole regions of UVA (315–399 nm), UVB (280–314 nm), and UVC (100–279). However, P. consimilis and P. dissimile’s leaves spectra have wider absorption peaks at the UVA region than P. aureum and S. sessilifolium . In the rhizomes (Fig. 5 D), P. divaricata, P. ursipes , and P. consimilis show a more intense absorption in the whole spectrum, but the effect is also more notorious in the UVA region. 3.5 Acclimatization effect on metabolite production Four out of the six ferns selected for acclimatization were used to determine metabolite concentrations: C. latum, N. nidulare, P. aureum , and S. attenuatum . We considered various characteristics to choose them, such as quinic and chlorogenic acids production of wild types, feasibility for pot cultivation (long-creeping rhizome species such as P. macrocarpa and P. loriceum need some characteristics from the wild and cannot grow in a standardized substratum), sample preservation and their availability ( N. crassifolium is a casual specie, and a population of this fern cannot be found). S attenuatum has not been included in the initial screening, although it is close to other Serpocaulon . Table 2 shows metabolite content for greenhouse-grown ferns. The production of metabolites differs significantly from wild ones (Fig. 2 ). Most acclimatized ferns increase quinic acid concentration, respecting their wild specimens. QA content increases from 0 to 3.64 (leaves), 2.29 (rhizomes) mg/g for C. latum to 13.94 mg/g for N. nidulare leaves, and 8.31 mg/g for P. aureum leaves. Rhizome QA content increases by 18% (13.31 the best acclimatized vs 11.29 mg/g) for P. aureum and 5% (17.97 vs 17.03 mg/g) for N. nidulare. CGA concentration is very similar for wild and acclimatized fern specimens (± 2 mg/g). The most important change is observed for C. latum leaves (2.42 mg/g in wild vs 9.85 mg/g acclimatized). Table 2 Metabolite concentration and Solar Protection Factor of acclimatized fern samples at different conditions based on dry mass (± standard deviations). Sample Shade QA CGA TPC SPF % (mg/g DM) (mg/g DM) (mg GAE/ g DM) C. latum leaves 50 ND ND ND ND 70 3.64 ± 0.08 9.85 ± 0.20 17.61 ± 0.13 ND C. latum rhizomes 50 ND ND ND ND 70 2.19 ± 0.05 0.21 ± 0.00 ND ND N. nidulare leaves 50 13.94 ± 0.20 1.39 ± 0.01 24.97 ± 0.94 ND 70 10.90 ± 0.30 1.70 ± 0.10 ND ND N. nidulare rhizomes 50 17.97 ± 0.43 0.42 ± 0.01 26.72 ± 1.29 3.42 ± 0.23 70 13.92 ± 0.98 1.85 ± 0.10 ND ND P. aureum leaves 50 8.31 ± 0.88 a 3.62 ± 0.19 a 38.35 ± 0.24 a ND 47.59 ± 3.26 b 70 8.29 ± 0.34 a 0.99 ± 0.02 a ND ND P. aureum rhizomes 50 13.31 ± 0.57 a 0.18 ± 0.00 a 24.69 ± 1.55 a 4.61 ± 1.39 a 8.79 ± 0.40 b 3.87 ± 2.98 b 70 4.38 ± 0.35 a 0 ND ND S. attenuatum leaves 50 ND ND ND ND 70 14.77 ± 1.46 2.50 ± 0.11 ND ND S. attenuatum rhizomes 50 ND ND ND ND 70 13.83 ± 0.61 1.54 ± 0.03 22.40 ± 0.65 4.42 ± 0.18 NOTES: QA quinic acid, CGA chlorogenic acid, TPC total polyphenolic content, SPF solar protection factor, DM dry mater, ND non determined. a high-altitude specimen, b low-altitude specimen TPC is increased with acclimatization for most ferns: C. latum increases from 1.28 to 17.61 mgGAE/gDM (leaves) and 1.04 to 24.97 (mgGAE/gDM), P. aureum from 41.18 to 47.59 mgGAE/gDM (leaves) and 7.89 to 24.69 mgGAE/gDM, and N. nidulare ’s leaves from 1.04 to 24.97 mgGAE/gDM (respecting best acclimatization condition). Only N. nidulare ’s rhizomes decrease TPC from 84.01 to 26.72 mgGAE/gDM. According to the literature, phenolic compounds can exhibit a UV-protectant mechanism, the same responsible for human skin protection. Then, higher altitudes and less shade can induce the production due to the higher exposition to UV sunlight 30 . Then, QA and TPC decrease with the increase in the shade, for all samples where 50 and 70% shade were recorded. On the other hand, many wild specimens showing high values of those compounds live on top of trees and places off the shade. Also, the SPF of N. nidulare’s rhizome extract decreases from 5.47 to 3.42 with acclimatization and increases from 1.69 to 4.61 for P. aureum ’s rhizomes, although those differences are not significant. 4. Conclusions We demonstrate wild ferns have an interesting potential for skin healing formulations. 19 out of 20 species included in this study had shown either some content of quinic and/or chlorogenic acid and/or antimicrobial activity. P. macrocarpa ’s and C. ursipes contain the highest quinic and chlorogenic acids found in wild ferns, respectively (30.09 and 8.5 mg/g). The rhizomes from wild samples of N. nidulare , N. crassifolium , and S. sessilifolium contain higher quinic acid than P. aureum which has been used commercially. Also 17 out of 19 species showed TPC higher than P. aureum rhizomes, and N. nidulare is the highest TPC with 84.10 mgGAE/gDM. Most wild ferns have higher SPF than P. aureum rhizomes, demonstrating their UV protectant capabilities. Ten out of twenty evaluated ferns demonstrated antimicrobial properties against the gram-positive bacteria S. aureus and S. epidermis -the extracts of N. crassifolium and S. loriceum are the most active ones. Fern’s acclimatization is feasible and increases QA content and TPC in most cases. Fewer shade conditions at greenhouses also increase QA concentration and TPC in N. nidulare , and P. pseudoaureum . Although we did not find a linear co-relationship between metabolites tested and UV protectant properties, this last effect is probably due to synergistic effects between some specific polyphenols and/or other polyunsaturated molecules. Declarations Acknowledgments This project has been financed by grant 0153-17 of the Institutional Founding for Scholar Development (FIDA, by its Spanish acronym) of the Universidad Nacional, Heredia, Costa Rica We thanks Ing. Montserrat Jimenez for assistance with figure 1, and Bach. Adrian Cerdas for the proofreading of the document. Author Contributions Conceptualization, funding acquisition, and supervision, G.RR., Y.CM., A.RA., and V.AV.; main experimental work, Y.SC., M.J.GB., J.PC., C.VR., J.A.RR.; writing, and editing P.JB. All authors contributed to reviewing and have read and agreed to the published version of the manuscript. Availability of Data and Materials Data will be available upon request. If data or materials are needed, please contact Victor Álvarez-Valverde, email: [email protected] Additional Information Competing Interests Statement The authors declare no conflict of interest. Ethical declarations The plants utilized in field experiments accomplish the institutional and national legislation. The protected vegetal material is authorized by the National Committee for the Management of Biodiversity (CONAGEBIO) of Costa Rica through the resolution R-CM-UNA-004-2018-OT References Greeshma, A. A. & Sridhar, K. R. in Medically important plant biomes: source of secondary metabolites (eds Dilfuza Egamberdieva & Antonio Tiezzi) 115–131 (Springer, 2019). Gonzalez, S., Alonso-Lebrero, J. L., Del Rio, R. & Jaen, P. Polypodium leucotomos extract: a nutraceutical with photoprotective properties. Drugs today 43 , 475–485 (2007). Gombau, L. et al. Polypodium leucotomos extract: antioxidant activity and disposition. Toxicol. In Vitro 20 , 464–471 (2006). Murbach, T. S. et al. A 28-day oral toxicology study of an aqueous extract of Polypodium leucotomos (Fernblock®). Toxicol. Rep. 4 , 494–501 (2017). Oh, J. H. et al. Antiphotoaging effects of 3, 5-dicaffeoyl-epi-quinic acid via inhibition of matrix metalloproteinases in UVB-irradiated human keratinocytes. Evid Based Complementary Altern. Med. 2020 (2020). Chen, Y. H. et al. Skin whitening capability of shikimic acid pathway compound. Eur. Rev. Med. Pharmacol. Sci. 20 , 1214–1220 (2016). Qian, Y., Qiu, X. & Zhu, S. Lignin: a nature-inspired sun blocker for broad-spectrum sunscreens. Green Chem. 17 , 320–324 (2015). Nestor, M. S., Berman, B. & Swenson, N. Safety and efficacy of oral Polypodium leucotomos extract in healthy adult subjects. J Clin Aesthet Dermatol 8 , 19 (2015). Dellacecca, E. R. et al. Antibiotics drive microbial imbalance and vitiligo development in mice. J. Invest. Dermatol. 140 , 676–687. e676 (2020). Ganju, P. et al. Microbial community profiling shows dysbiosis in the lesional skin of Vitiligo subjects. Sci. Rep. 6 , 1–10 (2016). Byrd, A., Yasmine, B. & Segre Julia, A. The human skin microbiome. Nat Rev Microbiol 16 , 143–155 (2018). Gleńsk, M. et al. Differing antibacterial and antibiofilm properties of Polypodium vulgare L. Rhizome aqueous extract and one of its purified active ingredients–osladin. J. Herb. Med. 17 , 100261 (2019). Parrado, C., Juarranz, A., Gilaberte, Y., Philips, N. & Gonzalez, S. in Cancer: Oxidative Stress and Dietary Antioxidants (ed Victor Preedy) 255–264 (Elsevier, 2014). Martin, S. L. et al. The occurrence of crassulacean acid metabolism in epiphytic ferns, with an emphasis on the Vittariaceae. Int J Plant Sci 166 , 623–630 (2005). Kluge, J. & Kessler, M. Influence of niche characteristics and forest type on fern species richness, abundance and plant size along an elevational gradient in Costa Rica. Plant. Ecol. 212 , 1109–1121 (2011). Guijarro-Real, C. et al. Growing conditions affect the phytochemical composition of edible wall rocket (Diplotaxis erucoides). Agronomy 9 , 858 (2019). Petkov, V. et al. Phytochemical Study and Biological Activity of Three Fern Species of the Asplenium Genus Growing in Bulgaria. Nat Prod J 12 , 82–90 (2022). Vega-López, B. et al. Phytonutraceutical evaluation of five varieties of tomato (Solanum lycopersicum) during ripening and processing. LWT, 113592 (2022). Balouiri, M., Sadiki, M. & Ibnsouda, S. K. Methods for in vitro evaluating antimicrobial activity: A review. J. Pharm. Anal. 6 , 71–79 (2016). Hudzicki, J. Kirby-Bauer Disk Diffusion Susceptibility Test Protocol , (2009). Zhang, Z., Zhang, B., Grishkewich, N., Berry, R. & Tam, K. C. Cinnamate-functionalized cellulose nanocrystals as UV‐shielding nanofillers in sunscreen and transparent polymer films. Advanced Sustainable Systems 3 , 1800156 (2019). Garcia, F. et al. Phenolic components and antioxidant activity of Fernblock (R), an aqueous extract of the aerial parts of the fern Polypodium leucotomos. Methods. Find. Exp. Clin. Pharmacol. 28 , 157–160 (2006). Dórea, J. G. & da Costa, T. H. M. Is coffee a functional food? Br. J. Nutr. 93 , 773–782 (2005). Yan, Y., Zhou, X., Guo, K., Zhou, F. & Yang, H. Use of Chlorogenic Acid against Diabetes Mellitus and Its Complications. J. Immunol. Res. 2020 (2020). Dvorakova, M. et al. Nutritional and antioxidant potential of fiddleheads from European ferns. Foods 10 , 460 (2021). Bai, J. et al. In vitro and in vivo characterization of the antibacterial activity and membrane damage mechanism of quinic acid against Staphylococcus aureus. J. Food. Saf. 38 , e12416 (2018). Lou, Z., Wang, H., Zhu, S., Ma, C. & Wang, Z. Antibacterial activity and mechanism of action of chlorogenic acid. J. Food Sci. 76 , M398-M403 (2011). Adamczak, A., Ożarowski, M. & Karpiński, T. M. Antibacterial activity of some flavonoids and organic acids widely distributed in plants. J. Clin. Med. 9 , 109 (2020). Sader, H. S., Farrell, D. J. & Jones, R. N. Antimicrobial susceptibility of Gram-positive cocci isolated from skin and skin-structure infections in European medical centres. Int. J. Antimicrob. Agents 36 , 28–32 (2010). Vetter, J. in Current Advances in Fern Research (ed Helena Fernandez) 305–327 (Springer, 2018). Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 18 Jan, 2024 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Major revision 29 Aug, 2023 Reviews received at journal 05 Jul, 2023 Reviewers agreed at journal 19 Jun, 2023 Reviews received at journal 11 Jun, 2023 Reviewers agreed at journal 06 Jun, 2023 Reviewers invited by journal 19 May, 2023 Editor assigned by journal 14 May, 2023 Editor invited by journal 06 Feb, 2023 Submission checks completed at journal 06 Feb, 2023 First submitted to journal 31 Jan, 2023 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2533922","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":173686269,"identity":"401bf93c-c3d4-4f37-8a23-ea7b1b25a9b5","order_by":0,"name":"Yaclyn Salazar-Chacon","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yaclyn","middleName":"","lastName":"Salazar-Chacon","suffix":""},{"id":173686270,"identity":"7db5c1b5-2d9a-438e-90ea-05209d1cb0a4","order_by":1,"name":"Maria José Gutierrez-Bolaños","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Maria","middleName":"José","lastName":"Gutierrez-Bolaños","suffix":""},{"id":173686271,"identity":"26e52330-966f-47a1-af00-123dd08177bb","order_by":2,"name":"Jimena Padilla-Cordero","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jimena","middleName":"","lastName":"Padilla-Cordero","suffix":""},{"id":173686272,"identity":"adbe482f-5231-46a7-b2ca-65ad65b50dac","order_by":3,"name":"Camilo Vidaurre-Rodriguez","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Camilo","middleName":"","lastName":"Vidaurre-Rodriguez","suffix":""},{"id":173686273,"identity":"d02d58df-9386-4b3b-bcc4-fa2c306b9d56","order_by":4,"name":"Yendry Carvajal-Miranda","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yendry","middleName":"","lastName":"Carvajal-Miranda","suffix":""},{"id":173686274,"identity":"03ec3ccf-b0df-4596-a625-12bdbf28d52b","order_by":5,"name":"Alexander Rojas-Alvarado","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Alexander","middleName":"","lastName":"Rojas-Alvarado","suffix":""},{"id":173686275,"identity":"718e8a64-7f8e-43f0-b3db-d10b37d750f1","order_by":6,"name":"Jorengeth Abad Rodríguez-Rodríguez","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jorengeth","middleName":"Abad","lastName":"Rodríguez-Rodríguez","suffix":""},{"id":173686276,"identity":"025293d1-7329-4097-a8d7-2b1259a63b7d","order_by":7,"name":"Pablo Jimenez-Bonilla","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABLUlEQVRIie2PMUvDQBTHXwg0y9WuEUq/gXASSAgOfpUEB5fYJUuGDBHBLnbvIPUr2KXzHQfpEu3aoahZOoUSF4mD4AuNAZOqq8P94I73uPvx3h9AIvmXEIUBdEjVYa2qlwyCXxVoKkrEIPlTqSlrVJTrnwVLe2Ash4P+0WjM0wLWA0tD5X0qhr1ITXMIn5qKfTN0+AQXM5PHM6rDxrCvlIiP58LXWcfQIfabCmUeFaRUVp6pUxDuvcAp3blwMZGJSzotZZlVynNmFU6l8I9b4d4x7W2vsqqnEBPD7xTRjbBgZO8Ue7LFLLTM4hmHEd0YtFT68bk7E8TXnbilWL0L/pIH8am5SI5fi2A9oEvB0yw8caeL0SzPw/Ziuzv+1laoeFpC/Sdsv0gkEonki0/4tHfDYx5r3AAAAABJRU5ErkJggg==","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Pablo","middleName":"","lastName":"Jimenez-Bonilla","suffix":""},{"id":173686277,"identity":"4621de22-0c01-4f83-8380-c2bf52e7d3b8","order_by":8,"name":"Gerardo Rodriguez-Rodriguez","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Gerardo","middleName":"","lastName":"Rodriguez-Rodriguez","suffix":""},{"id":173686278,"identity":"f8daf518-587f-452f-b737-9475635f68ba","order_by":9,"name":"Victor Alvarez-Valverde","email":"","orcid":"","institution":"Universidad Nacional (UNA)","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Victor","middleName":"","lastName":"Alvarez-Valverde","suffix":""}],"badges":[],"createdAt":"2023-01-31 13:14:28","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2533922/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2533922/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-023-50281-3","type":"published","date":"2024-01-18T15:01:23+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":32611204,"identity":"4fb3a025-c624-46be-a4ee-cfc4298ebdb4","added_by":"auto","created_at":"2023-02-07 20:31:50","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":265023,"visible":true,"origin":"","legend":"\u003cp\u003eSampling locations for biological material. (A) wild ferns. (B) acclimatized ferns. Maps constructed using Leaflet Map tiles by Stamen Design, CC BY 3.0 OpenStreetMap Contributors.\u003c/p\u003e\n\u003cp\u003eNOTES: \u003csup\u003e# Location\u003c/sup\u003e (altitude in meters above the medium level of the sea mamsl): species at the location. \u003csup\u003e1\u003c/sup\u003e (300 mamsl): \u003cem\u003eCampyloneurum amphostenon, Campyloneurum latum, Polypodium plesiosorum \u003c/em\u003evar\u003cem\u003e. rubicundum, Polypodium ursipes, Serpocaulon dissimile\u003c/em\u003e*\u003cem\u003e, Serpocaulon fraxinifolium, Serpocaulon sessilifolium\u003c/em\u003e; \u003csup\u003e2\u003c/sup\u003e (540 mamsl): \u003cem\u003eSerpocaulon triseriale\u003c/em\u003e; \u003csup\u003e3\u003c/sup\u003e (1413 mamsl): \u003cem\u003eNiphidium nidulare\u003c/em\u003e; \u003csup\u003e4\u003c/sup\u003e (1475 mamsl): \u003cem\u003ePolypodium colpodes\u003c/em\u003e; \u003csup\u003e5\u003c/sup\u003e (2227 mamsl): \u003cem\u003ePecluma divaricata, Pleopeltis montigena, Pleopeltis myriolepis, Serpocaulon loriceum\u003c/em\u003e; \u003csup\u003e6\u003c/sup\u003e (1092 mamsl): \u003cem\u003eCampyloneurum xalapense, Niphidium crassifolium, Pecluma consimilis, Polypodium sp, Serpocaulon dissimile\u003c/em\u003e; \u003csup\u003e7\u003c/sup\u003e (2180 mamsl): \u003cem\u003eCampyloneurum amphostenon, Campyloneurum latum, Polypodium plesiosorum \u003c/em\u003evar. \u003cem\u003erubicundum\u003c/em\u003e; \u003cem\u003ePolypodium ursipes, Serpocaulon dissimile, Serpocaulon fraxinifolium, Serpocaulon sessilifolium, Polypodium ursipes, Pleopeltis macrocarpa\u003c/em\u003e; \u003csup\u003e8\u003c/sup\u003e (2985 mamsl): \u003cem\u003ePolypodium ursipes\u003c/em\u003e; \u003csup\u003e9\u003c/sup\u003e (3088 mamsl) \u003cem\u003eSerpocaulon fraxinifolium (l), Serpocaulon sessilifolium (l);\u003c/em\u003e \u003csup\u003e10\u003c/sup\u003e (2115 mamsl): \u003cem\u003ePhlebodium aureum\u003c/em\u003e; \u003csup\u003e11\u003c/sup\u003e (635 mamsl) \u003cem\u003ePhlebodium aureum\u003c/em\u003e; \u003csup\u003e12\u003c/sup\u003e (830 mamsl): \u003cem\u003eCampyloneurum latum\u003c/em\u003e; \u003csup\u003e13\u003c/sup\u003e (2185 mamsl): \u003cem\u003ePhlebodium aureum\u003c/em\u003e; \u003csup\u003e14\u003c/sup\u003e (12 mamsl): \u003cem\u003eSerpocaulon attenuatum\u003c/em\u003e; \u003csup\u003e15\u003c/sup\u003e (2620 mamsl): \u003cem\u003eNiphidium nidulare\u003c/em\u003e. *\u003cem\u003eS. dissimile\u003c/em\u003e low altitude\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2533922/v1/56ee2cb905a9e459529a81f9.jpg"},{"id":32611205,"identity":"dc4cecf6-b5ac-4157-8f24-62686ebf7c64","added_by":"auto","created_at":"2023-02-07 20:31:50","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":839384,"visible":true,"origin":"","legend":"\u003cp\u003eSelected metabolite concentration of 21 Costa Rican wild ferns samples (dry base). (A) Quinic acid content of fern leaves. (B) Chlorogenic acid content of fern leaves. (C) Quinic acid content of fern rhizomes. (D) Chlorogenic acid content of fern rhizomes. Error bars represent standard deviation. Dashed and dotted lines represent \u003cem\u003ePhlebodium aureum\u003c/em\u003e leaves and rhizome concentration, respectively.\u003c/p\u003e\n\u003cp\u003e* Low altitude specimen\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2533922/v1/25b63e39a63211fe23d56ffa.jpg"},{"id":32611663,"identity":"b9cd48a1-aa2a-4797-9d4d-8fca6fce5331","added_by":"auto","created_at":"2023-02-07 20:39:50","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":418926,"visible":true,"origin":"","legend":"\u003cp\u003eTotal polyphenolic content of 19 Costa Rican wild ferns samples (dry base) determined by means of Folin-Ciocalteau method. (A) Fern leaves. (B) Fern rhizomes. Error bars represent standard deviation. Dashed and dotted lines represent \u003cem\u003ePhlebodium aureum\u003c/em\u003e leaves and rhizome concentration, respectively.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2533922/v1/fcb37ae6340c86f018324799.jpg"},{"id":32611202,"identity":"69c45a49-e4ef-4f78-b5be-6da9f2a95e95","added_by":"auto","created_at":"2023-02-07 20:31:50","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":787362,"visible":true,"origin":"","legend":"\u003cp\u003eAntibacterial potential of 21 wild fern extracts. (A) Leaves extracts against \u003cem\u003eS. aureus. \u003c/em\u003e(B) Leaves extracts against \u003cem\u003eS. epidermidis. \u003c/em\u003e(C) Rhizome extracts against \u003cem\u003eS. aureus. \u003c/em\u003e(D) Rhizome extracts against \u003cem\u003eS. epidermidis. \u003c/em\u003e50μL of 1000 μg/mL of Penicillin/Streptomycin\u003cem\u003e \u003c/em\u003ewere used as positive control.\u003cem\u003e \u003c/em\u003eError bars represent standard deviation.\u003c/p\u003e\n\u003cp\u003e* Low altitude specimen\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2533922/v1/dff24801d4094412948b38b4.jpg"},{"id":32611206,"identity":"fe3db0ba-7779-40d7-afb9-3152a2a09a7a","added_by":"auto","created_at":"2023-02-07 20:31:50","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":676733,"visible":true,"origin":"","legend":"\u003cp\u003eSolar Protection Factor of 19 Costa Rican wild ferns samples (dry base) at a concentration equivalent to the found in the original dry material, and ultraviolet absorption spectra of selected samples. (A) SPF of leaves samples. (B) SPF of rhizomes samples. (C) UV spectrum of selected leaves samples. (D) UV spectrum of selected rhizome samples. For A \u0026amp; B, error bars represent standard deviation, and dashed and dotted lines represent \u003cem\u003ePhlebodium aureum\u003c/em\u003e leaves and rhizome SPF, respectively. C and D spectra were recorded at a constant concentration for all samples.\u003c/p\u003e","description":"","filename":"5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2533922/v1/840a0508e36301e012caa544.jpg"},{"id":49978868,"identity":"a3ad403d-a16b-4c7f-9276-5700c553e7d3","added_by":"auto","created_at":"2024-01-22 15:09:44","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1254704,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2533922/v1/4e2237a9-aef3-4465-9acf-37077e636ba5.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Exploration of photoprotective and antibiotic activity of 20 wild Polypodiaceae ferns from Costa Rica","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eFerns are primitive non-flowering vascular plants, and many of them have shown medicinal properties. Several ferns have been studied as bio-herbicides, hepatoprotectives, cytotoxic, antihyperglycemic, trypanocidal, antimicrobial, antinociceptive, and immunomodulatory\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Nutritional, nutraceutical and pharmacological utilization are still limited at a commercial scale, with some exceptions, such as kalawalla (\u003cem\u003ePhlebodium aureum\u003c/em\u003e formerly known as \u003cem\u003ePolypodium leucotomos\u003c/em\u003e). The extract of \u003cem\u003eP. aureum\u003c/em\u003e has been used to treat skin diseases since the \u0026lsquo;80s, and there are many commercial preparations based on the hydrosoluble extract of their rhizomes, such as Fernblock\u0026reg;, Heliocare\u0026reg;, Difur\u0026reg;, and others. \u003cem\u003eP. aureum\u003c/em\u003e extracts have been commercialized mainly in Europe despite this plant is endemic to Central America \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eOral or topic formulations containing \u003cem\u003eP. aureum\u003c/em\u003e extract are recognized for their photoprotectant bioactivity and their strong antioxidant properties, which have been related to their naturally occurring phenolic compounds such as \u003cem\u003ep\u003c/em\u003e-coumaric, ferulic, caffeic, vanillic, and chlorogenic acid \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. Commercial aqueous extract of the leaves of \u003cem\u003eP. aureum (\u003c/em\u003emarketed as Fernblock\u0026reg;, Fernplus\u0026reg;, Fernmed\u0026reg;, and Fernage\u0026reg;) contain a standardized 0.6\u0026ndash;1.3% total phenolic compounds concentration and 0.4\u0026ndash;0.9% quinic acid concentration \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. Mechanisms of action of these commercial products involve free radical scavenging and regulation of several genes, such as metalloproteinases \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e, tyrosinase inhibition \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e, and others. Also, natural phenolic compounds are known for absorbing UV radiation, similar to synthetic compounds, such as homosalate or dibenzalacetone do so. Then fern extracts are an excellent alternative for bio-based wide-spectrum sun blockers and sunscreen\u003csup\u003e\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. Also, fern extracts have been found active to prevent UV damage during clinical trials\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e, preventing sunburns, and subsequently skin cancer.\u003c/p\u003e \u003cp\u003eFern-derived nutraceutical products are an alternative for some skin disorders with unavailable treatment options, such as eczema or vitiligo. Also, skin microbiome imbalance and traditional antibiotic utilization contribute to developing and speeding up skin depigmentation \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. For example, \u003cem\u003eStaphylococcus\u003c/em\u003e spp. is more abundant on the skin of patients with vitiligo \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e,\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. Some aqueous fern rhizome extracts showed antibacterial properties against gram-negative bacteria (such as \u003cem\u003eEscherichia coli\u003c/em\u003e) and gram-positive (such as \u003cem\u003eS. aureus\u003c/em\u003e) \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eOther Central American \u003cem\u003ePolypodiaceae\u003c/em\u003e and \u003cem\u003ePteridophyta\u003c/em\u003e ferns have similar metabolic pathways to \u003cem\u003eP. aureum\u003c/em\u003e\u003csup\u003e\u003cem\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/em\u003e\u003c/sup\u003e, and also have been used for the same applications in traditional medicine\u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Although some wild ferns are not feasible to reproduce in a greenhouse, there is an enormous potential for the discovery and commercialization of medicinal ferns in Central America\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e. Also, growing conditions such as climate, altitude, light incidence, and substrate composition can be used to improve or standardize metabolite production\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e,\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. Therefore, we hypothesize that other ferns can have similar or superior activity compared to \u003cem\u003eP. aureum\u003c/em\u003e, in terms of UV protection and metabolite concentration. Some of them can be potentially more active or more economic to produce than kalawalla is. In this work, we explored wild and acclimatized ferns in terms of amounts of quinic acid, chlorogenic acid, and total polyphenolic compounds, as potential alternatives to treat skin disorders.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"2. Materials And Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003e2.1 Microorganism strains\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eThe strains utilized in this study were \u003cem\u003eStaphylococcus aureus\u003c/em\u003e ATCC 25923, \u003cem\u003eStaphylococcus epidermidis\u003c/em\u003e ATCC 12228, \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e ATCC 9027, and \u003cem\u003eEscherichia coli\u003c/em\u003e ATCC 25922.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n\u003ch2\u003e2.2 Fern samples\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eTwenty wild native fern species were collected from the forest at 13 locations. Figure\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e(A) summarizes the identified species and their location. Both rhizomes and leaves were collected for each plant, oven-dried for 72 h at 40\u0026deg;C, and ground to 1 mm.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n\u003ch2\u003e2.3 Quinic and chlorogenic acid determination\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003e0.5 g dry material (rhizome or leaves) was extracted in 1.5 mL 75% methanol. Afterwards, it was filtered through a 0.2 \u0026micro;m membrane. The extract was dried into an analytic evaporator ZipVap Zanntek (from Glas-Col\u0026reg;, Terre Haute, IN) at 40\u0026deg;C under nitrogen. Then, it was reconstituted in 500 \u0026micro;L 75% methanol. The reconstituted extract was analyzed by HPLC, using a solvent gradient described in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. The column utilized is an Acclaim 120 (C-18 (4,6 \u0026times;150) mm, 5 \u0026micro;m) (from Thermo Scientific Dionex, CA) at 35\u0026deg;C. A diode array detector was used to detect quinic acid (QA) and chlorogenic acid (CGA) at 215 and 325 nm, respectively, against 0-2000 mg/L QA and 0-600 mg/L CGA standard curves.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eStarting, interception and ending points for linear gradients of mobile phase, utilized to quantify quinic and chlorogenic acid through HPLC.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePoint\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eTime (min)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eFlow rate\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eAcetonitrile\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMetanol\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e0.1%TFA\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eStarting\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e90%\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIntersection\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80%\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIntersection\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e20%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70%\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIntersection\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e40%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50%\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIntersection\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80%\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eEnding\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e90%\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n\u003ch2\u003e2.4 Total polyphenolic content (TPC) quantification\u003c/h2\u003e\n\u003cp\u003e0.1 g of each sample was mixed with 3 mL of 75% methanol and sonicated for 10 min. Then, the mixture was centrifuged for 5 min, and the methanolic extract was collected. The remaining solid was extracted two additional times following the same procedure. All the extracts were combined and diluted to complete 10 mL (total volume). Then, Folin-Ciocalteu\u0026rsquo;s colorimetric method was used to determine total phenolic content, as described in our previous work\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e, using gallic acid as standard. Each sample has three replicates. Both \u003cem\u003eS. dissimile\u003c/em\u003e specimens were combined in equal amounts for this analysis.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n\u003ch2\u003e2.5 Antibiotic activity test\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eIn two replicates, we followed the agar well diffusion method \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e, a modification of the Kirby-Bauer susceptibility protocol \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. These experiments were conducted on Mueller Hinton agar plates. 100 \u0026micro;L of each microorganism at 0,5 of McFarlan scale (1,5x 10\u003csup\u003e8\u003c/sup\u003e UFC/mL) in NaCl 0.85% were plated into a sterile 90x15mm petri dish containing 25 mL of Mueller Hinton agar. Then, four 6 mm wells were made in the agar at the same distance, three wells were filled with 50\u0026micro;L of 0.1g/mL fern extract, and the fourth well was filled with 50 \u0026micro;L of 50%v/v acetone (as negative control). Additionally, for the positive control, the same procedure was carried out, but 3 well were filled with 50\u0026micro;L of 1000 \u0026micro;g/mL of Penicillin/Streptomycin (from GIBCO, NY), and the fourth well was filled with 50 \u0026micro;L of 50%v/v acetone (as negative control). Plates were incubated for 24 h at 35\u0026deg;C, and inhibition rings were measured. Two replicates were used for each condition. Microbial activity was calculated according to (1).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Equ1\" class=\"Equation\"\u003e\n\u003cdiv id=\"FileID_Equ1\" class=\"mathdisplay\"\u003e$$\\text{%}RPI=\\frac{({D}_{sample}-{D}_{negative control})}{\\left({D}_{positive control}\\right)}x100$$\u003c/div\u003e\n\u003cdiv class=\"EquationNumber\"\u003e1\u003c/div\u003e\n\u003c/div\u003e\n\u003cp\u003e%\u003cem\u003eRPI\u003c/em\u003e is the Relative Percentage of Inhibition, and \u003cem\u003eD\u003c/em\u003e\u003csub\u003e\u003cem\u003esample\u003c/em\u003e\u003c/sub\u003e, \u003cem\u003eD\u003c/em\u003e\u003csub\u003e\u003cem\u003enegative control\u003c/em\u003e\u003c/sub\u003e, and \u003cem\u003eD\u003c/em\u003e\u003csub\u003e\u003cem\u003epositive control\u003c/em\u003e\u003c/sub\u003e are the diameters of the inhibition zones indicated in the subscripts.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n\u003ch2\u003e2.6 Solar protection factor (SPF) determination\u003c/h2\u003e\n\u003cp\u003eThe same extracts prepared for TPC analysis were diluted 100 times, and the UV-Vis spectra were recorded for each sample repetition between 190\u0026ndash;800 nm in a spectrophotometer Evolution 350 (from Thermo Scientific, Massachusetts, USA). Data were exported and analyzed according to Mansur Eq.\u0026nbsp;2\u003csup\u003e1\u003c/sup\u003e. SPFs were adjusted to the corresponding concentration found in the dry material (leaves or rhizome), and the average of 3 commercial sunscreens (SPF 50) was used as reference: Nivea Sun\u0026reg;, Eucerin\u0026reg;, and Vicky face\u0026reg; sunscreens. 3 repetitions had been used.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n\u003ch2\u003e2.7 Acclimatization of selected ferns\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eSix fern species were selected to grow under controlled conditions to study the acclimatization effect over metabolite production. \u003cem\u003eCampyloneurum latum\u003c/em\u003e T. Moore, \u003cem\u003eNiphidium nidulare\u003c/em\u003e (Rosenst.) Lellinger, \u003cem\u003ePhlebodium pseudoaureum\u003c/em\u003e (Cav.) Lellinger, \u003cem\u003eSerpocaulon attenuatum\u003c/em\u003e (C. Presl) A.R. Sm., \u003cem\u003eS. sessilifolium\u003c/em\u003e (Desv.) A.R. Sm. y \u003cem\u003eS. triseriale\u003c/em\u003e (Sw.) A.R. Sm. were collected from the wild at the locations mentioned in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e(B). The samples were reproduced at Macho River Biological Station, located at 9\u0026deg;46'00\" N, 83\u0026deg;46'00\" W, a 1600 mamsl, in Oros\u0026iacute;, Cartago, Costa Rica. The reported temperature range at the location is 18,3\u0026ndash;26,5\u0026deg;C (average 22,3\u0026deg;C), and the annual precipitation is 3120 mm. 68 plants of each specie were distributed under 50% and 70% shade (covered with saran fabric) in 1\u0026ndash;3 L plant pots containing a substrate composed by: peat (1/4), organic fertilizer (1/4), grounded coconut fiber (1/4) and coconut shell chunks (1/4). They were harvested after a year, processed, and analyzed using the same methods described for wild ferns.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"3. Results And Discussion","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n\u003ch2\u003e3.1 Content of quinic and chlorogenic acids of wild ferns\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e summarizes the metabolite content of leaves and rhizome of the 20 wild ferns included in this study. \u003cem\u003ePolypodium leucotomos\u003c/em\u003e is a synonym for \u003cem\u003ePhlebodium pseudoaureum\u003c/em\u003e (sensu stricto) or \u003cem\u003ePhlebodium aureum\u003c/em\u003e (sensu lato).\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eQuinic acid was present in eleven leaf extracts and eight rhizome extracts. Notwithstanding the commercial product elaborated from the rhizome extract, we decided to include the leaves, considering that metabolites could be distributed all over the ferns. Interestingly, quinic acid was detected in leaves of \u003cem\u003ePecluma consimilis\u003c/em\u003e (Mett.) M.G. Price, \u003cem\u003ePolypodium colpodes\u003c/em\u003e Kunze, \u003cem\u003eP. plesiosorum\u003c/em\u003e Kunze and \u003cem\u003eSerpocaulon dissimile\u003c/em\u003e (L.) A.R. Sm., but it was not detected in the rhizomes of those species. Also, quinic acid was detected in both rhizomes and leaves of \u003cem\u003eNiphidium crassifolium\u003c/em\u003e (L.) Lellinger, \u003cem\u003ePleopeltis macrocarpa\u003c/em\u003e (Bory ex Willd.) Kaulf., \u003cem\u003eP. montingena\u003c/em\u003e (Maxon) A.R. Sm. \u0026amp; Tejero, \u003cem\u003eP. myriolepis\u003c/em\u003e (Christ) A.R. Sm. \u0026amp; Tejero, \u003cem\u003eSerpocaulon sessilifolium\u003c/em\u003e (Desv.) A.R. Sm. and \u003cem\u003eS. triseriale\u003c/em\u003e (Sw.) A.R. Sm., but the concentration of leaves was higher in most of them, except for \u003cem\u003eN. crassifolium\u003c/em\u003e and \u003cem\u003eS. sessilifolium\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eQuinic acid was not detected in \u003cem\u003eP. aureum\u003c/em\u003e leaves, but it accounts for 11.2 mg/g in the rhizome. Figure\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(A) contains the quinic acid concentration found on fern leaves. Three species showed a quinic acid content 56\u0026ndash;67% lower, respecting our reference value (\u003cem\u003eP. aureum\u003c/em\u003e rhizome): \u003cem\u003eP. consimilis\u003c/em\u003e, \u003cem\u003eP. myriolepis\u003c/em\u003e, and \u003cem\u003eP. plesiosorum\u003c/em\u003e. Another five species have ranged from similar levels to 30% higher quinic acid content (than \u003cem\u003eP. aureum\u003c/em\u003e rhizome): \u003cem\u003eN. crassifolium\u003c/em\u003e, \u003cem\u003eP. montigena\u003c/em\u003e, \u003cem\u003eP. colpodes\u003c/em\u003e, \u003cem\u003eS. dissimile\u003c/em\u003e, and \u003cem\u003eS. sessilifolium\u003c/em\u003e. Finally, the leaves of three fern species contain between 1.7 to 2.7 fold-up respecting \u003cem\u003eP. aureum\u003c/em\u003e rhizome: \u003cem\u003eP. macrocarpa\u003c/em\u003e, \u003cem\u003eS. dissimile\u003c/em\u003e, and \u003cem\u003eS. triseriale\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(C) shows quinic acid in rhizomes. Four species (\u003cem\u003eP. macrocarpa\u003c/em\u003e, \u003cem\u003eP. montigena P. myriolepis\u003c/em\u003e, and \u003cem\u003eS. triseriale\u003c/em\u003e) contain lower quinic acid than \u003cem\u003eP. aureum\u003c/em\u003e. But also, three species contain 1.5\u0026ndash;2.3 higher quinic acid than \u003cem\u003eP. aureum\u003c/em\u003e: \u003cem\u003eN. crassifolium\u003c/em\u003e, \u003cem\u003eN. nidulare\u003c/em\u003e, and \u003cem\u003eS. sessilifolium.\u003c/em\u003e According to the literature, an analysis of Fernblock\u0026reg;, the commercial extract from \u003cem\u003eP. aureum\u003c/em\u003e accounts for 70 g/L quinic acid. The three species with higher quinic acid concentration are potentially a more economical source of this metabolite \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eAlso, phenolic compounds account for 1% of commercial Fernblock\u0026reg; \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e, composed mainly of 3,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, vanillic acid, caffeic acid, \u003cem\u003ep-\u003c/em\u003ecoumaric acid, ferulic acid, 4-hydroxycinnamoyl-quinic acid, and five isomers of chlorogenic acid \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. Chlorogenic acid is a dietary polyphenol \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e, linked with activity against inflammation, skin diseases, and others \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e23\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e, so we had included its quantification in this study. Chlorogenic acid was present in leaves, but not in rhizomes of \u003cem\u003eP. pseudoaureum\u003c/em\u003e, \u003cem\u003eP. macrocarpa, P colpodes, P. plesiororum\u003c/em\u003e, \u003cem\u003eP. ursipes\u003c/em\u003e Moritz ex C. Chr. and \u003cem\u003eS. triseriale\u003c/em\u003e. Four species \u003cem\u003eC. amphostenon\u003c/em\u003e (Kunze ex Klotzsch) F\u0026eacute;e, \u003cem\u003eC. latum, P. myriolepis\u003c/em\u003e, and \u003cem\u003ePolypodium sp\u003c/em\u003e, contain chlorogenic acid in both leaves and rhizomes.\u003c/p\u003e\n\u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(D) shows the chlorogenic acid content of wild ferns included in this study. Although chlorogenic acid is reported as a phenolic component of \u003cem\u003eP. aureum\u003c/em\u003e commercial formulations \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e, we did not find a detectable concentration on the wild rhizome, because of its very low concentration (if present). However, five rhizome extracts contain chlorogenic acid: \u003cem\u003eC. amphostenon, C. latum\u003c/em\u003e, \u003cem\u003eN. nidulare\u003c/em\u003e, \u003cem\u003eP. myriolepis\u003c/em\u003e, and \u003cem\u003ePolypodium sp.\u003c/em\u003e, ranging from 0.8\u0026ndash;4.7 mg/g DM. Figure\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e(B) show chlorogenic acid content in leaves. \u003cem\u003eP. aureum\u003c/em\u003e\u0026rsquo;s leaves contain around 3.7 mg/g chlorogenic acid, and another seven ferns contain around 0.8\u0026ndash;3.7 mg/g of this metabolite (\u003cem\u003eS. triseriale, Polypodium sp., P. plesiosorum, P. colpodes, P. macrocarpa, C. latum\u003c/em\u003e, and \u003cem\u003eC. amphostenon\u003c/em\u003e). Two fern leaves contain significantly higher chlorogenic acid content (7.9\u0026ndash;8.5 g/L): \u003cem\u003eP. ursipes\u003c/em\u003e and \u003cem\u003eP. myriolepsis\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eFrom all these results, we highlight the high quinic acid content in \u003cem\u003eN. nidulare\u003c/em\u003e and \u003cem\u003eS. sessilifolium\u003c/em\u003e rhizomes, and \u003cem\u003eP. macrocapa\u003c/em\u003e leaves. Other elements to consider in the future, to determine the feasibility of industrialization of these wild ferns, are reproduction, culturing, and toxicity. \u003cem\u003eP. leucotomos\u003c/em\u003e extract is safe for human consumption. Some other ferns are known to contain ptaquiloside (a nor-sesquiterpene), and sesquiterpenes exhibiting mutagenic, teratogenic, clastogenic, and carcinogenic activities \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n\u003ch2\u003e3.2 Total Polyphenolic Content of wild ferns\u003c/h2\u003e\n\u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e summarizes the TPC of the leaves and rhizomes of the ferns. TPC in leaves (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA) is found the highest for \u003cem\u003eP. consimilis\u003c/em\u003e (74.2 mgGAE/gDM). Two other fern leaves contain TPC considerably higher than \u003cem\u003eP. aureum\u003c/em\u003e leaves (41.2 mgGAE/gDM): \u003cem\u003eS. dissimile\u003c/em\u003e (55.3 mgGAE/gDM) and \u003cem\u003eP. divaricata\u003c/em\u003e (56.8 mgGAE/gDM). Other five fern species contain similar TPC (\u0026plusmn;\u0026thinsp;6.6 mg mgGAE/gDM) than \u003cem\u003eP. aureum\u003c/em\u003e leaves: \u003cem\u003eP. montigena\u003c/em\u003e (43.9 mgGAE/gDM), \u003cem\u003eP. macrocarpa\u003c/em\u003e (42.64 mgGAE/gDM), \u003cem\u003eS. sessilifolium\u003c/em\u003e (38.84 mgGAE/gDM), \u003cem\u003eP. colpodes\u003c/em\u003e (35.8 mgGAE/gDM), and \u003cem\u003ePolypodium\u003c/em\u003e sp. (34.6 mgGAE/gDM).\u003c/p\u003e\n\u003cp\u003eInterestingly, most of both leaves\u0026rsquo; samples (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA) and rhizome\u0026rsquo;s samples (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eB) contain higher TPC than \u003cem\u003eP. aureum\u003c/em\u003e rhizome (commercially used), except \u003cem\u003eN. nidulare\u0026rsquo;s\u003c/em\u003e and \u003cem\u003eC. latum\u0026rsquo;s\u003c/em\u003e leaves; and, \u003cem\u003eC. amphostenon\u003c/em\u003e\u0026rsquo;s, \u003cem\u003eP. colpodes\u003c/em\u003e\u0026rsquo;s and \u003cem\u003eC. latum\u003c/em\u003e\u0026rsquo;s rhizomes despite the last two are virtually the same TPC.\u003c/p\u003e\n\u003cp\u003eIn rhizomes, seven species have shown the highest TPC particularly high: \u003cem\u003eN. nidulare\u003c/em\u003e (84.10 mgGAE/gDM), \u003cem\u003eP. divaricata\u003c/em\u003e (71.73 mgGAE/gDM), \u003cem\u003eP. ursipes\u003c/em\u003e (68.17 mgGAE/gDM), \u003cem\u003eN. crassifolium\u003c/em\u003e (70.00 mgGAE/gDM), \u003cem\u003eP. myriolepis\u003c/em\u003e (56.87 mgGAE/gDM), \u003cem\u003eP. consimilis\u003c/em\u003e (54.58 mgGAE/gDM), and \u003cem\u003eP. montigena\u003c/em\u003e (40.00 mgGAE/gDM).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n\u003ch2\u003e3.3 Antibacterial properties of fern extracts\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e summarizes the relative percent of inhibition of the extracts. The leaves \u003cem\u003eof N. crassifolium, P. aureum, P. macrocarpa, P. montigena, P. myriolepis, P. colpodes, S. dissimile\u003c/em\u003e, and \u003cem\u003eS. triseriale\u003c/em\u003e were found active against both \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eS. epidermis\u003c/em\u003e, showing similar relative inhibition to the positive control (around 40\u0026ndash;62%). All the other extracts from leaves do not possess antimicrobial activity.\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eThe activity of rhizome extracts can be divided into three groups. The strongest relative inhibition against \u003cem\u003eS. aureus\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eC) was found in the range of 65\u0026ndash;80% for the extracts of \u003cem\u003eN. crassifolium\u003c/em\u003e, \u003cem\u003eP. montigena\u003c/em\u003e, and \u003cem\u003eS. loriceum\u003c/em\u003e. The first two extracts show a similar inhibition against \u003cem\u003eS. epidermis\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eD) too. An intermediate range of the strength of relative inhibition (45\u0026ndash;65%) was observed for \u003cem\u003eN. nidulare\u003c/em\u003e, \u003cem\u003eP. macrocarpa\u003c/em\u003e, and \u003cem\u003eS. triseriale\u003c/em\u003e, against both bacteria (\u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eS. epidermis)\u003c/em\u003e. Extracts from \u003cem\u003eP. montigena\u003c/em\u003e also showed an intermediate inhibition against \u003cem\u003eS. epidermis\u003c/em\u003e. Finally, there is a group with a moderate relative inhibition (\u0026lt;\u0026thinsp;45%), including the rhizomic extracts from \u003cem\u003eC. xalapense\u003c/em\u003e F\u0026eacute;e, \u003cem\u003eP. consimilis\u003c/em\u003e, and \u003cem\u003eS. fraxinifolium\u003c/em\u003e, with antimicrobial against \u003cem\u003eS. aureum\u003c/em\u003e, and the extracts from \u003cem\u003eP. ursipes\u003c/em\u003e against both \u003cem\u003eS. aureum\u003c/em\u003e and \u003cem\u003eS. epidermis\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eQuinic and chlorogenic acids are reported to be antibacterial against several strains of \u003cem\u003eS. aureus\u003c/em\u003e, \u003cem\u003eE. coli\u003c/em\u003e, some \u003cem\u003ebacillus\u003c/em\u003e, and others \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e26\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. However, in our results, none of both compounds showed any activity against the strains evaluated here under the same conditions tested for the fern extracts (data not shown). Yet, some other compounds such as flavonoids (flavones, flavonols, flavanones), and organic acids (aliphatic and aromatic acids) are known for being antimicrobial \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e. Therefore, we evaluated the crude extracts of the wild ferns against the gram-negative: \u003cem\u003eP. aeruginosa\u003c/em\u003e and \u003cem\u003eE. coli\u003c/em\u003e, and the gram-positive \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eS. epidermidis\u003c/em\u003e. Our results show that the ferns possess antimicrobial against the gram-positive bacteria tested but not against the gram-negative ones. (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Remarkably, most skin infections involve gram-positive bacteria, and antibiotic resistance is occurring more frequently in them \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n\u003ch2\u003e3.4 Solar Protection Factor (SPF) of fern extracts\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eFigures \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eA and \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eB is showing the SPF for 19 fern species included in this study. Interestingly, almost all the samples have shown higher SPF than \u003cem\u003eP. aureum\u003c/em\u003e rhizomes, except just \u003cem\u003eN. nidulare\u003c/em\u003e\u0026rsquo;s leaves. The highest SPF in leaves (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eA) is \u003cem\u003eP. consimiles\u003c/em\u003e (29), \u003cem\u003eS. sessilifolium\u003c/em\u003e (28), and \u003cem\u003eS. dissimile\u003c/em\u003e (28). Also, highest SPF in rhizomes is \u003cem\u003eP. divaricata\u003c/em\u003e (18), \u003cem\u003eP. ursipes\u003c/em\u003e (15), and \u003cem\u003eP. consimiles\u003c/em\u003e (13). We did not find a correlationship between TPC, quinic acid, or chlorogenic acid concentration and SPF. UV absorption is probably influenced by more than one compound, and according to Mensul\u0026rsquo;s equation, compounds with high absorptivity at 300\u0026ndash;305 nm are the most important contributors to SPF. It is very likely, most compounds responsible for UV absorption are polyphenols, although, some of them could have maximum absorption at other wavelengths or low molar absorptivities, and this is why there is no correlationship between SPF and TPC.\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eSelected ultraviolet spectra containing \u003cem\u003eP. aureum\u003c/em\u003e and the samples with the highest SPF are shown in Figs.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eC and \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eD. All the samples cover the whole regions of UVA (315\u0026ndash;399 nm), UVB (280\u0026ndash;314 nm), and UVC (100\u0026ndash;279). However, \u003cem\u003eP. consimilis\u003c/em\u003e and \u003cem\u003eP. dissimile\u0026rsquo;s\u003c/em\u003e leaves spectra have wider absorption peaks at the UVA region than \u003cem\u003eP. aureum\u003c/em\u003e and \u003cem\u003eS. sessilifolium\u003c/em\u003e. In the rhizomes (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eD), \u003cem\u003eP. divaricata, P. ursipes\u003c/em\u003e, and \u003cem\u003eP. consimilis\u003c/em\u003e show a more intense absorption in the whole spectrum, but the effect is also more notorious in the UVA region.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n\u003ch2\u003e3.5 Acclimatization effect on metabolite production\u003c/h2\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n\u003cp\u003eFour out of the six ferns selected for acclimatization were used to determine metabolite concentrations: \u003cem\u003eC. latum, N. nidulare, P. aureum\u003c/em\u003e, and \u003cem\u003eS. attenuatum\u003c/em\u003e. We considered various characteristics to choose them, such as quinic and chlorogenic acids production of wild types, feasibility for pot cultivation (long-creeping rhizome species such as \u003cem\u003eP. macrocarpa\u003c/em\u003e and \u003cem\u003eP. loriceum\u003c/em\u003e need some characteristics from the wild and cannot grow in a standardized substratum), sample preservation and their availability (\u003cem\u003eN. crassifolium\u003c/em\u003e is a casual specie, and a population of this fern cannot be found). S attenuatum has not been included in the initial screening, although it is close to other \u003cem\u003eSerpocaulon\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eTable\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e shows metabolite content for greenhouse-grown ferns. The production of metabolites differs significantly from wild ones (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). Most acclimatized ferns increase quinic acid concentration, respecting their wild specimens. QA content increases from 0 to 3.64 (leaves), 2.29 (rhizomes) mg/g for \u003cem\u003eC. latum\u003c/em\u003e to 13.94 mg/g for \u003cem\u003eN. nidulare\u003c/em\u003e leaves, and 8.31 mg/g for \u003cem\u003eP. aureum\u003c/em\u003e leaves. Rhizome QA content increases by 18% (13.31 the best acclimatized vs 11.29 mg/g) for \u003cem\u003eP. aureum\u003c/em\u003e and 5% (17.97 vs 17.03 mg/g) for \u003cem\u003eN. nidulare.\u003c/em\u003e CGA concentration is very similar for wild and acclimatized fern specimens (\u0026plusmn;\u0026thinsp;2 mg/g). The most important change is observed for \u003cem\u003eC. latum\u003c/em\u003e leaves (2.42 mg/g in wild vs 9.85 mg/g acclimatized).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eMetabolite concentration and Solar Protection Factor of acclimatized fern samples at different conditions based on dry mass (\u0026plusmn;\u0026thinsp;standard deviations).\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cspan class=\"Underline\"\u003eSample\u003c/span\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cspan class=\"Underline\"\u003eShade\u003c/span\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cspan class=\"Underline\"\u003eQA\u003c/span\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cspan class=\"Underline\"\u003eCGA\u003c/span\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cspan class=\"Underline\"\u003eTPC\u003c/span\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cspan class=\"Underline\"\u003eSPF\u003c/span\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(mg/g DM)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(mg/g DM)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(mg GAE/ g DM)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eC. latum\u003c/em\u003e leaves\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e9.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eC. latum\u003c/em\u003e rhizomes\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eN. nidulare\u003c/em\u003e leaves\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e24.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.94\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10.90\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eN. nidulare\u003c/em\u003e rhizomes\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e17.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e26.72\u0026thinsp;\u0026plusmn;\u0026thinsp;1.29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.98\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eP. aureum\u003c/em\u003e leaves\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.88 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e38.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e47.59\u0026thinsp;\u0026plusmn;\u0026thinsp;3.26 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eP. aureum\u003c/em\u003e rhizomes\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.31\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.00 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e24.69\u0026thinsp;\u0026plusmn;\u0026thinsp;1.55 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.61\u0026thinsp;\u0026plusmn;\u0026thinsp;1.39 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.40 \u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e3.87\u0026thinsp;\u0026plusmn;\u0026thinsp;2.98 b\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35 \u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eS. attenuatum\u003c/em\u003e leaves\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e14.77\u0026thinsp;\u0026plusmn;\u0026thinsp;1.46\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eS. attenuatum\u003c/em\u003e rhizomes\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eND\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e13.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.61\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e22.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.65\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.42\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003ctfoot\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\"\u003eNOTES: QA quinic acid, CGA chlorogenic acid, TPC total polyphenolic content, SPF solar protection factor, DM dry mater, ND non determined. \u003csup\u003ea\u003c/sup\u003e high-altitude specimen, \u003csup\u003eb\u003c/sup\u003e low-altitude specimen\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tfoot\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTPC is increased with acclimatization for most ferns: \u003cem\u003eC. latum\u003c/em\u003e increases from 1.28 to 17.61 mgGAE/gDM (leaves) and 1.04 to 24.97 (mgGAE/gDM), \u003cem\u003eP. aureum\u003c/em\u003e from 41.18 to 47.59 mgGAE/gDM (leaves) and 7.89 to 24.69 mgGAE/gDM, and \u003cem\u003eN. nidulare\u003c/em\u003e\u0026rsquo;s leaves from 1.04 to 24.97 mgGAE/gDM (respecting best acclimatization condition). Only \u003cem\u003eN. nidulare\u003c/em\u003e\u0026rsquo;s rhizomes decrease TPC from 84.01 to 26.72 mgGAE/gDM.\u003c/p\u003e\n\u003cp\u003eAccording to the literature, phenolic compounds can exhibit a UV-protectant mechanism, the same responsible for human skin protection. Then, higher altitudes and less shade can induce the production due to the higher exposition to UV sunlight\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. Then, QA and TPC decrease with the increase in the shade, for all samples where 50 and 70% shade were recorded. On the other hand, many wild specimens showing high values of those compounds live on top of trees and places off the shade. Also, the SPF of \u003cem\u003eN. nidulare\u0026rsquo;s\u003c/em\u003e rhizome extract decreases from 5.47 to 3.42 with acclimatization and increases from 1.69 to 4.61 for \u003cem\u003eP. aureum\u003c/em\u003e\u0026rsquo;s rhizomes, although those differences are not significant.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. Conclusions","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eWe demonstrate wild ferns have an interesting potential for skin healing formulations. 19 out of 20 species included in this study had shown either some content of quinic and/or chlorogenic acid and/or antimicrobial activity. \u003cem\u003eP. macrocarpa\u003c/em\u003e\u0026rsquo;s and \u003cem\u003eC. ursipes\u003c/em\u003e contain the highest quinic and chlorogenic acids found in wild ferns, respectively (30.09 and 8.5 mg/g). The rhizomes from wild samples of \u003cem\u003eN. nidulare\u003c/em\u003e, \u003cem\u003eN. crassifolium\u003c/em\u003e, and \u003cem\u003eS. sessilifolium\u003c/em\u003e contain higher quinic acid than \u003cem\u003eP. aureum\u003c/em\u003e which has been used commercially. Also 17 out of 19 species showed TPC higher than \u003cem\u003eP. aureum\u003c/em\u003e rhizomes, and \u003cem\u003eN. nidulare\u003c/em\u003e is the highest TPC with 84.10 mgGAE/gDM. Most wild ferns have higher SPF than \u003cem\u003eP. aureum\u003c/em\u003e rhizomes, demonstrating their UV protectant capabilities. Ten out of twenty evaluated ferns demonstrated antimicrobial properties against the gram-positive bacteria \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eS. epidermis\u003c/em\u003e -the extracts of \u003cem\u003eN. crassifolium\u003c/em\u003e and \u003cem\u003eS. loriceum\u003c/em\u003e are the most active ones. Fern\u0026rsquo;s acclimatization is feasible and increases QA content and TPC in most cases. Fewer shade conditions at greenhouses also increase QA concentration and TPC in \u003cem\u003eN. nidulare\u003c/em\u003e, and \u003cem\u003eP. pseudoaureum\u003c/em\u003e. Although we did not find a linear co-relationship between metabolites tested and UV protectant properties, this last effect is probably due to synergistic effects between some specific polyphenols and/or other polyunsaturated molecules.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis project has been financed by grant 0153-17 of the Institutional Founding for Scholar Development (FIDA, by its Spanish acronym) of the Universidad Nacional, Heredia, Costa Rica We thanks Ing. Montserrat Jimenez for assistance with figure 1, and Bach. Adrian Cerdas for the proofreading of the document.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor Contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization, funding acquisition, and supervision, G.RR., Y.CM., A.RA., and V.AV.; main experimental work, Y.SC., M.J.GB., J.PC., C.VR., J.A.RR.; writing, and editing P.JB. All authors contributed to reviewing and have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of Data and Materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData will be available upon request. If data or materials are needed, please contact Victor \u0026Aacute;lvarez-Valverde, email: [email protected]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAdditional Information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests Statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical declarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe plants utilized in field experiments accomplish the institutional and national legislation. The protected vegetal material is authorized by the National Committee for the Management of Biodiversity (CONAGEBIO) of Costa Rica through the resolution R-CM-UNA-004-2018-OT\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eGreeshma, A. A. \u0026amp; Sridhar, K. 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Antibacterial activity of some flavonoids and organic acids widely distributed in plants. J. Clin. Med. \u003cb\u003e9\u003c/b\u003e, 109 (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSader, H. S., Farrell, D. J. \u0026amp; Jones, R. N. Antimicrobial susceptibility of Gram-positive cocci isolated from skin and skin-structure infections in European medical centres. Int. J. Antimicrob. Agents \u003cb\u003e36\u003c/b\u003e, 28\u0026ndash;32 (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVetter, J. in \u003cem\u003eCurrent Advances in Fern Research\u003c/em\u003e (ed Helena Fernandez) 305\u0026ndash;327 (Springer, 2018).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"quinic acid, vitiligo, fern, chlorogenic acid, skin, antibiotic, bacteria","lastPublishedDoi":"10.21203/rs.3.rs-2533922/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2533922/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eSkin disorders affect millions of people all over the world. There are limited options to treat dermal illnesses such as vitiligo, psoriasis, and atopic dermatitis (eczema). Central American ferns are a potential source of bioactive metabolites against those diseases. Currently, Polypodium leucotomos Poir. (Phlebodium aureum (L.) J. Sm. synonym) is the only one being commercially utilized for this purpose. In this work, we evaluated the concentration of the skin bioactive compounds: quinic and chlorogenic acid, in the extract of 20 wild ferns from Costa Rica. We also evaluated the antimicrobial capabilities of the raw extracts of wild ferns and the sun protection factor (SPF) of the extracts. We found 19 out of 20 have either an important concentration of the compounds mentioned above or antimicrobial properties. Also, most samples result in higher SPF than P. aureum\u0026rsquo;s rhizome. We also have studied the fern acclimatization, at different shading conditions, finding a significant influence of the culturing conditions on metabolite production. After acclimatization. So far, we demonstrate that various ferns included in this study are a potential source of treatments for skin conditions.\u003c/p\u003e","manuscriptTitle":"Exploration of photoprotective and antibiotic activity of 20 wild Polypodiaceae ferns from Costa Rica","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-02-07 20:31:45","doi":"10.21203/rs.3.rs-2533922/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2023-08-29T08:01:21+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2023-07-05T06:04:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"f36b9431-1669-49f7-b68c-f0fb005b273f","date":"2023-06-19T16:13:14+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2023-06-12T01:38:26+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"30980994-a77c-4d63-a44c-325ed9f2d59c","date":"2023-06-06T05:27:41+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2023-05-19T14:46:03+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-05-14T06:28:03+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2023-02-06T10:04:07+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2023-02-06T09:31:13+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2023-01-31T13:11:16+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"6bb63552-61dd-43c4-9f8c-3d62fa1937c3","owner":[],"postedDate":"February 7th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":18979433,"name":"Physical sciences/Chemistry/Medicinal chemistry"},{"id":18979434,"name":"Physical sciences/Chemistry/Chemical biology/Natural products"},{"id":18979435,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2024-01-22T15:06:34+00:00","versionOfRecord":{"articleIdentity":"rs-2533922","link":"https://doi.org/10.1038/s41598-023-50281-3","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2024-01-18 15:01:23","publishedOnDateReadable":"January 18th, 2024"},"versionCreatedAt":"2023-02-07 20:31:45","video":"","vorDoi":"10.1038/s41598-023-50281-3","vorDoiUrl":"https://doi.org/10.1038/s41598-023-50281-3","workflowStages":[]},"version":"v1","identity":"rs-2533922","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2533922","identity":"rs-2533922","version":["v1"]},"buildId":"FbvkV6FR0MCFSLy54lSbu","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}