Clusia nemorosa, popularly known as "pororoca" is used in folk medicine as anti-inflammatory and analgesic. However, until the moment, do not has scientific records to confirm this observation. Thus, motivated by this information, in the present study we evaluated the anti-inflammatory effect of the hexane extract obtained from the bark and leaf of Clusia nemorosa using different experimental models. In the model of acute inflammation induced by carrageenan, the extract of the bark and leaf were able to inhibit the neutrophil accumulation in the pleural cavity after 4 h of stimulation. In addition, both extracts significantly reduced the levels of TNF-alpha in pleural fluid of animals stimulated with carrageenan. However, in the in vitro studies, only the hexane extract of the leaf was able to demonstrate a direct effect on these cells by inhibiting the chemotaxis of purified human neutrophils stimulated with CXCL1. This inhibition in the cell motility do not seems to be related to events of death, because the analysis by flow cytometry revealed that incubation of human neutrophils with the extracts was not able to interfere with the viability of these cells. In the model of allergic inflammation, only the hexane extract of the bark suppressed the recruitment of eosinophils into the pleural cavity 24 h after antigen challenge in actively sensitized animals. Analysis of gene expression of proteins associated with allergies (IL-5 and CCL11) showed that the bark extract also significantly inhibited the production of messenger RNA for IL-5 and CCL11. In the model of chronic inflammation with granuloma formation induced by cotton implants, we observed that the hexane extract of the bark showed the greatest ability to inhibit the formation of granulomatous tissue. Thus, this set of results supports the popular use of the extract obtained from Clusia nemorosa, and indicates that different parts (bark and leaf) of this plant can become a potential source of substances with antiinflammatory properties. / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Clusia nemorosa, conhecida popularmente como pororoca , é empregada na medicina popular como antiinflamatório e analgésico, porém não há registros científicos que confirmem esta observação. Assim, motivados por esta informação, no presente estudo avaliamos o efeito antiinflamatório do extrato hexânico obtido da casca e da folha de Clusia nemorosa utilizando diferentes modelos experimentais. No modelo de inflamação aguda induzida por carragenina, os extratos da casca e da folha foram capazes de inibir o acúmulo de neutrófilos na cavidade pleural de camundongos no tempo de 4 h. Além disso, ambos os extratos reduziram de maneira significativa os níveis de TNF-alfa no fluido pleural de animais estimulados com carragenina. Entretanto, in vitro apenas o extrato hexânico da folha foi capaz de demonstrar um efeito direto sobre estas células por inibir a quimiotaxia de neutrófilos humanos purificados estimulados por CXCL1. Esta inibição na motilidade celular parece não ser relacionada a eventos de morte, pois a análise realizada por citometria de fluxo revelou que a incubação dos neutrófilos humanos com os extratos não foi capaz de interferir com a viabilidade destas células. No modelo de inflamação alérgica, somente o extrato hexânico da casca suprimiu o recrutamento de eosinófilos para o espaço pleural após desafio antigênico em animais ativamente sensibilizados no tempo de 24 h. Análise da expressão gênica de proteínas relacionadas ao processo alérgico (IL-5 e CCL11) revelou que o extrato da casca também inibiu de maneira significativa a produção de RNA mensageiros para IL-5 e CCL11. No modelo de inflamação crônica com formação de granuloma induzido por implantes de algodão, observamos que o extrato hexânico da casca foi o que apresentou maior capacidade de inibição da formação do tecido granulomatoso. Assim, este conjunto de resultados sustenta o uso popular do extrato obtido de Clusia nemorosa, além de indicar que diferentes partes (casca e folha) desta planta podem torna-se uma potencial fonte de substâncias com propriedades antiinflamatórias.

Clusia nemorosa is distributed throughout Brazil and its extract is widely used in popular medicine to treat different diseases. Although the popular use of extract of C. nemorosa, the scientific reported on their pharmacological and phytochemical analysis are scarce. In the present study, we aimed to evaluate the antinociceptive and antiinflammatory effect of hexane extract of Clusia nemorosa-derived stem bark (EHC). To evaluate the antinociceptive effect of EHC classical models of the study of pain were used, such as the abdominal writhing test induced by acetic acid, formalin test and hot plate test. In order to evaluate anti-inflammatory effect of EHC, the model of paw edema induced by carrageenan were used. Rota-rod testing and toxicological analysis in vivo and in vitro were performed. Mices submitted to abdominal writhing test, was observed a significant reduction in nociceptive response induced by acetic acid, and these effects persisted for up to 2 h after treatment. Moreover, it is important emphasizing that the EHC did not induce changes in motor function of animals when the rota-rod test was performed. In order to investigate the possible mechanism of action of EHC, the animals were pretreated with naloxone (5mg/kg), yohimbine (1 mg/kg), atropine (5 mg/kg), metoclopramide (1 mg/kg), haloperidol (2 mg/kg) and L -NAME (20 mg/kg). Only when used metoclopramide, a serotoninergic and dopaminergic antagonist receptor, it was observed an inhibition of the antinociceptivo effect of EHC, suggesting that the serotoninergic pathway may be involved in the action of the EHC, as haloperidol, a dopaminergic antagonist did not reverse the antinociception of EHC. In addition, EHC did not induce changes in the nociceptive response of animals when assessed by the hot plate test, discarding the action in the central nervous system. Then, when the animals were treated with EHC and submitted to the formalin test, it was observed that only the second phase of this test was inhibited by the EHC, indicating inhibitory actions on pain of EHC from inflammatory origin. To evaluate possible toxic effects of EHC, animals were treated for 7 consecutive days. Then after treatment, were not observed changes in behavior, blood cellularity and cytotoxicity in vitro, which ruled out possible of toxic effects in these preliminary tests. Through analysis of gas chromatography coupled with mass spectrum, the major component present in the extract was identified, friedelin, who presented antinociceptive effects on pain and inflammation of neurogenic origin, as evidenced by the formalin test. Although this model of pain induced by formalin, it was observed that the antinociceptive effects friedelin were maintained when the EHC was administered orally. In addition, friedelin inhibited the paw edema induced by carrageenan. Lastly, in vitro assays, this triterpene did not affect cell viability as measured by MTT test. Together, our results support the popular use of this plant based on their anti-inflammatory and antinociceptive effects. In addition, we report the first time the presence of pentacyclic triterpenoid, friedelin on hexane extract of Clusia nemorosa-derived stem bark, triterpenoid which is possibly responsible for the pharmacological effects evaluated. / Conselho Nacional de Desenvolvimento Científico e Tecnológico / A espécie Clusia nemorosa é bem distribuída pelo território brasileiro e seus extratos são amplamente utilizados pela medicina popular para tratar diferentes doenças. Mesmo sendo comum o consumo dos extratos de C. nemorosa (EHC), os registros científicos sobre as propriedades farmacológicas e análises fitoquímicas desta espécie ainda mostram-se escassos. Assim, no presente estudo objetivamos avaliar o efeito antinociceptivo e anti-inflamatório do extrato hexânico obtido a partir das cascas do caule de Clusia nemorosa. Para traçar o perfil antinociceptivo foram utilizados os modelos de contorção abdominal induzido por ácido acético, formalina e o teste de placa quente. Com propósito de avaliar o perfil anti-inflamatório utilizamos o teste de edema de pata induzido por carragenina. Para melhor compreender as ações do EHC, submetemos os animais tratados com o extrato ao teste de rota-rod e a análises toxicológicas in vivo e in vitro. No ensaio de contorção abdominal foi observada uma redução na resposta nociceptiva induzida por ácido acético, sendo estes efeitos persistentes por até 2 h após tratamento com EHC. Através do teste de rota rod, verificamos que o EHC não induziu alterações na motricidade dos animais. Com propósito de investigar o possível mecanismo de ação deste extrato os animais foram pré-tratados com naloxona (5 mg/kg), ioimbina (1 mg/kg), atropina (5 mg/kg), metoclopramida (1 mg/kg), haloperidol (2 mg/kg) e LNAME (20 mg/kg). Apenas quando utilizado a metoclopramida, antagonista de receptores serotoninérgicos e dopaminérgicos, ocorreu inibição do efeito antinociceptivo do EHC, sugerindo que as vias serotoninérgicas possam estar envolvidas nas ações do extrato, pois o haloperidol, um antagonista dopaminérgico não reverteu a antinocicepção do EHC. Em adição, o tratamento com EHC não induziu alterações na resposta nociceptiva dos animais quando avaliados pelo teste da placa quente, descartando possíveis ações no Sistema Nervoso Central. Em seguida, quando os animais tratados com extrato foram submetidos ao teste de formalina, foi observado que apenas a segunda fase deste teste foi inibida, indicando ações inibitórias sobre a dor de origem inflamatória, revertida pelo pré-tratamento com metoclopramida. Quando os animais foram submetidos ao tratamento por 7 dias consecutivos com o EHC não foram observadas alterações no comportamento, na celularidade do sangue e citotoxicidade in vitro, o que descartou possíveis efeitos tóxicos nestes testes preliminares. Através de análise de cromatografia gasosa acoplada a espectro de massa, foi identificado o componente majoritário presente no extrato, a friedelina (1 e 10 mg/kg; i.p.), que apresentou efeitos atinociceptivos na dor de origem neurogênica e inflamatória, evidenciado pelo teste de formalina. Ainda neste modelo, foi observado que a friedelina manteve seus efeitos antinociceptivos na dor de origem inflamatória quando administrado por via oral. Além disso, a friedelina inibiu o edema de pata induzido por carragenina. Em adição, em ensaios in vitro, este triterpeno não alterou a viabilidade celular. Em conjunto, nossos resultados sustentam o uso popular desta planta tendo por base seus efeitos antinociceptivos e anti-inflamatórios. Além disso, relatamos pela primeira vez a presença do triterpenóide pentacíclico, friedelina, na casca de C. nemorosa, sendo este triterpenóide possivelmente responsável pelos efeitos farmacológicos avaliados.

Buchenwälder sind die großflächigste potenziell natürliche Vegetationsform Deutschlands und ein nach EU-FFH-Richtlinie besonders zu schützender Biotoptyp. Eine hohe Naturnähe ist auch in Wirtschaftswäldern (WiWald) notwendig, um die typischen Lebensgemeinschaften naturnaher Wälder langfristig zu erhalten, doch mangelt es an praktikablen/verifizierten Indikatoren, wie die nutzungsbedingte Abweichung vom Naturzustand ermittelt werden kann. In >100 Jahre alten und ~40 ha großen Tiefland-Buchenwäldern (Mecklenburg-Vorpommern/Brandenburg) wurde anhand von 13 WiWäldern, vier seit <20 Jahren (k20) und drei seit >50 Jahren (r50) unbewirtschafteten Beständen den folgenden Fragen nachgegangen: Wie groß sind die strukturellen, vegetationskundlichen und carabidologischen Unterschiede zwischen bewirtschafteten, kurz- und langfristig unbewirtschafteten Buchenwäldern? Gibt es strukturelle Indikatoren und quantitative Größen zur Abschätzung des Einflusses forstlicher Bewirtschaftung auf die Biozönosen von Tiefland-Buchenwäldern? In Probekreisen (Pk) von 500 m² an Rasterpunkten (100 m x 100 m) wurden strukturelle und in Pk von 314 m² vegetationskundliche Daten erhoben. In fünf Pk/Bestand wurde jeweils eine Barberfalle über die Vegetationsperiode installiert. Ganzflächig wurden die Verteilung der Waldentwicklungsphasen (WEP)und zusätzlich zu den Pk-Aufnahmen hektarweise Sonderstrukturen aufgenommen. U. a. wurden folgende Sonderstrukturen aufgenommen: Zunderschwamm, Kronen- und Zwieselbrüche, Ersatzkronen, Blitzrinnen, Risse/Spalten, Höhlen, Mulmkörper/-taschen. Diese naturschutzfachlich wichtigen Sonderstrukturen wurden aus den Habitatansprüchen der typischen Buchenwaldfauna abgeleitet.Es konnten große Unterschiede zwischen WiWald und r50-Flächen (v. a. >100 Jahre unbewirtschafteten Flächen) aufgezeigt werden. Die k20-Flächen unterscheiden sich nicht wesentlich vom WiWald. Die Anzahl verschiedener WEP/ha und WEP-Patches/ha liegt in den r50-Flächen signifikant höher als im WiWald. Der Holzvorrat der r50-Flächen liegt mit ~600 m³/ha (Terminal- ~800 m³/ha, Zerfallsphase 450 m³/ha) deutlich höher als im WiWald. Charakteristisch für die r50-Flächen ist das Vorkommen von in ihrer Vitalität eingeschränkten Bäume ab 80 cm BHD und ein inhomogeneres Lichtmosaik im Bestand. Die Stammqualitäten (u. a. Astigkeit) in r50-Flächen unterscheiden sich kaum von denen in WiWald. In den r50-Flächen kommt bedeutend mehr Totholz (>142 m³/ha) als im WiWald (max. 34 m³/ha) vor. Im WiWald können Stubben dominieren. Verschiedene Totholzqualitäten sind im WiWald nur unvollständig vorhanden. Etwa 40 % des Totholzes besitzt keine Totholznachbarn (r50-Flächen: <2 %) und die Lichtverhältnisse am Totholz sind nicht so vielfältig (wenig sonnenexponiert und wenig gering besonnt). In den >100 Jahre unbewirtschafteten Flächen kommen ~12 Sonderstrukturtypen mit >200 Sonderstrukturen/ha vor. 19 von 20 Sonderstrukturen sind im WiWald signifikant seltener und 11 Sonderstrukturen sind als Naturnähe-Indikatoren geeignet.Vegetation: In der Krautschicht sind höhere Deckungsgrade, mehr (lichtanzeigende) Arten, weniger Waldarten und eine höhere Diversität zu verzeichnen. Im WiWald wird u. a. das Vorkommen von Calamagrostis epigeios, Impatiens parviflora und Rubus idaeus gefördert. Stark gefährdete Moosarten sind im WiWald seltener als in den Referenzwäldern, da sie vor allem auf liegendem Totholz und auf den Stammanläufen vorkommen. Carabiden: Im WiWald gibt es weniger Individuen und Biomasse von mesophilen Waldarten und eine geringere Anzahl von flugunfähigen Individuen. Als Indikatoren für naturnahe Tiefland-Buchenwälder können die drei Arten Carabus glabratus, C. hortensis und Cychrus caraboides bezeichnet werden. Indikatoren: Es wurden Zielgrößen für 29 Struktur-Indikatoren für naturnahe Wälder vorgeschlagen. Für WiWälder wurden gesonderte Zielgrößen festgelegt, die die nutzungsbedingte, nicht zu vermeidende Abweichung vom Naturzustand berücksichtigen. / Beech forests are the most important natural vegetation type of Germany,and they are included in annex II of the EU-FFH-Directive,which requests nature conservation for the listed habitat types.High naturalness is necessary in managed forests (w-sites) to maintain the typical biocoenosis of forests near nature. But there is a lack of practicable/verified indicators to determine the degree of alteration managed forests have compared to natural forests. In >100 year old and ~40 ha big lowland beech forests in Mecklenburg-Vorpommern and Brandenburg, 13 w-sites, 4 study sites which are unmanaged since <20 years (k-sites) and 3 sites which are unmanaged since >50 years (r50-sites) were investigated to answer these questions: What the differences are between w-, k- and r-sites according to forest structure, vegetation and carabids? Are there valid structural indicators with thresholds to assess the impact of forestry use on the biocoenosis of lowland beech forests? At grid points(distance 100 mx 100 m),on circular sample plots (SP) of 500 m² the structural data and on SP of 314 m² the vegetation was investigated. At five SP/study site a pitfall trap was installed during the entire vegetation period. On the whole study site the distribution of forest development phases (FDP) was mapped, and on full one ha plots the special structures were investigated. The following special structures were mapped e.g. Fomes fomentarius trees, crown and crotch breakage, substitute crowns, lightning shakes,gutters/rifts, cavities, mould and bark bag. These special structures have been derived from the habitat needs of the typical beech forest fauna.The results revealed tremendous differences between w- and r50-sites. The k-sites show no clear differences to the managed sites.In the r50-sites, the number of different FDP/ha and FDP units/ ha is significant higher than in w-sites. The timber stock of the r50-sites is ~600 m³/ha (terminal phase ~800 m³/ha, decay phase ~450³/ha). A characteristic feature of the r50-sites is the occurrence of trees with 80 cm bhd or more with reduced vitality. The timber trunk) qualities of r-sites differ only slightly from managed stands. In the r50-sites the dead wood volume (>142 m³/ha) is much higher than in the w-sites (max. 34 m³/ha). Many different features of dead wood occur only fragmentary within w-sites. About 40 % of the dead wood objects have no "dead wood neighbour" (r50-sites: <2 %), and the light distribution is much less diverse. In >100 years unmanaged r-sites ~12 different types of special structures and 200 single special structures occur per ha. 19 out of 20 special structures are significantly less frequent in w-sites; 11 special structures are specifically valuable to be used as naturalness indicators.Vegetation: In the herb layer occur higher coverage values, more (light-indicating) species, but only few species indicating ancient forests and a higher diversity index value. In w-sites, the occurrence of e. g. Calamagrostis epigeios, Impatiens parviflora and Rubus idaeus is supported. reduced. Threatened moss species are rare in w-sites compared to r-sites, since they mainly grow on laying dead wood, which is rare in forests in use, and on inclined/rough-barked stem bases. Ground beetles: The forestry use of lowland beech forests leads to less individuals and lower biomass of so-called mesophilous forest species. Furthermore, the number of flightless individuals is lower. As proper indicators for near-natural lowland beech forests, the three species Carabus glabratus, C. hortensis und Cychrus caraboides could be identified. Indicators: 29 structural indicators were identified and thresholds were given. But even in lowland beech forests managed in a conservation-friendly way, these target values for near-natural and natural forests are unlikely to be reached. Therefore, for w-sites special threshold values have been defined, which consider the inevitable difference between managed and natural forests.

Buchenwälder sind die großflächigste potenziell natürliche Vegetationsform Deutschlands und ein nach EU-FFH-Richtlinie besonders zu schützender Biotoptyp. Eine hohe Naturnähe ist auch in Wirtschaftswäldern (WiWald) notwendig, um die typischen Lebensgemeinschaften naturnaher Wälder langfristig zu erhalten, doch mangelt es an praktikablen/verifizierten Indikatoren, wie die nutzungsbedingte Abweichung vom Naturzustand ermittelt werden kann. In >100 Jahre alten und ~40 ha großen Tiefland-Buchenwäldern (Mecklenburg-Vorpommern/Brandenburg) wurde anhand von 13 WiWäldern, vier seit <20 Jahren (k20) und drei seit >50 Jahren (r50) unbewirtschafteten Beständen den folgenden Fragen nachgegangen: Wie groß sind die strukturellen, vegetationskundlichen und carabidologischen Unterschiede zwischen bewirtschafteten, kurz- und langfristig unbewirtschafteten Buchenwäldern? Gibt es strukturelle Indikatoren und quantitative Größen zur Abschätzung des Einflusses forstlicher Bewirtschaftung auf die Biozönosen von Tiefland-Buchenwäldern? In Probekreisen (Pk) von 500 m² an Rasterpunkten (100 m x 100 m) wurden strukturelle und in Pk von 314 m² vegetationskundliche Daten erhoben. In fünf Pk/Bestand wurde jeweils eine Barberfalle über die Vegetationsperiode installiert. Ganzflächig wurden die Verteilung der Waldentwicklungsphasen (WEP)und zusätzlich zu den Pk-Aufnahmen hektarweise Sonderstrukturen aufgenommen. U. a. wurden folgende Sonderstrukturen aufgenommen: Zunderschwamm, Kronen- und Zwieselbrüche, Ersatzkronen, Blitzrinnen, Risse/Spalten, Höhlen, Mulmkörper/-taschen. Diese naturschutzfachlich wichtigen Sonderstrukturen wurden aus den Habitatansprüchen der typischen Buchenwaldfauna abgeleitet.Es konnten große Unterschiede zwischen WiWald und r50-Flächen (v. a. >100 Jahre unbewirtschafteten Flächen) aufgezeigt werden. Die k20-Flächen unterscheiden sich nicht wesentlich vom WiWald. Die Anzahl verschiedener WEP/ha und WEP-Patches/ha liegt in den r50-Flächen signifikant höher als im WiWald. Der Holzvorrat der r50-Flächen liegt mit ~600 m³/ha (Terminal- ~800 m³/ha, Zerfallsphase 450 m³/ha) deutlich höher als im WiWald. Charakteristisch für die r50-Flächen ist das Vorkommen von in ihrer Vitalität eingeschränkten Bäume ab 80 cm BHD und ein inhomogeneres Lichtmosaik im Bestand. Die Stammqualitäten (u. a. Astigkeit) in r50-Flächen unterscheiden sich kaum von denen in WiWald. In den r50-Flächen kommt bedeutend mehr Totholz (>142 m³/ha) als im WiWald (max. 34 m³/ha) vor. Im WiWald können Stubben dominieren. Verschiedene Totholzqualitäten sind im WiWald nur unvollständig vorhanden. Etwa 40 % des Totholzes besitzt keine Totholznachbarn (r50-Flächen: <2 %) und die Lichtverhältnisse am Totholz sind nicht so vielfältig (wenig sonnenexponiert und wenig gering besonnt). In den >100 Jahre unbewirtschafteten Flächen kommen ~12 Sonderstrukturtypen mit >200 Sonderstrukturen/ha vor. 19 von 20 Sonderstrukturen sind im WiWald signifikant seltener und 11 Sonderstrukturen sind als Naturnähe-Indikatoren geeignet.Vegetation: In der Krautschicht sind höhere Deckungsgrade, mehr (lichtanzeigende) Arten, weniger Waldarten und eine höhere Diversität zu verzeichnen. Im WiWald wird u. a. das Vorkommen von Calamagrostis epigeios, Impatiens parviflora und Rubus idaeus gefördert. Stark gefährdete Moosarten sind im WiWald seltener als in den Referenzwäldern, da sie vor allem auf liegendem Totholz und auf den Stammanläufen vorkommen. Carabiden: Im WiWald gibt es weniger Individuen und Biomasse von mesophilen Waldarten und eine geringere Anzahl von flugunfähigen Individuen. Als Indikatoren für naturnahe Tiefland-Buchenwälder können die drei Arten Carabus glabratus, C. hortensis und Cychrus caraboides bezeichnet werden. Indikatoren: Es wurden Zielgrößen für 29 Struktur-Indikatoren für naturnahe Wälder vorgeschlagen. Für WiWälder wurden gesonderte Zielgrößen festgelegt, die die nutzungsbedingte, nicht zu vermeidende Abweichung vom Naturzustand berücksichtigen. / Beech forests are the most important natural vegetation type of Germany,and they are included in annex II of the EU-FFH-Directive,which requests nature conservation for the listed habitat types.High naturalness is necessary in managed forests (w-sites) to maintain the typical biocoenosis of forests near nature. But there is a lack of practicable/verified indicators to determine the degree of alteration managed forests have compared to natural forests. In >100 year old and ~40 ha big lowland beech forests in Mecklenburg-Vorpommern and Brandenburg, 13 w-sites, 4 study sites which are unmanaged since <20 years (k-sites) and 3 sites which are unmanaged since >50 years (r50-sites) were investigated to answer these questions: What the differences are between w-, k- and r-sites according to forest structure, vegetation and carabids? Are there valid structural indicators with thresholds to assess the impact of forestry use on the biocoenosis of lowland beech forests? At grid points(distance 100 mx 100 m),on circular sample plots (SP) of 500 m² the structural data and on SP of 314 m² the vegetation was investigated. At five SP/study site a pitfall trap was installed during the entire vegetation period. On the whole study site the distribution of forest development phases (FDP) was mapped, and on full one ha plots the special structures were investigated. The following special structures were mapped e.g. Fomes fomentarius trees, crown and crotch breakage, substitute crowns, lightning shakes,gutters/rifts, cavities, mould and bark bag. These special structures have been derived from the habitat needs of the typical beech forest fauna.The results revealed tremendous differences between w- and r50-sites. The k-sites show no clear differences to the managed sites.In the r50-sites, the number of different FDP/ha and FDP units/ ha is significant higher than in w-sites. The timber stock of the r50-sites is ~600 m³/ha (terminal phase ~800 m³/ha, decay phase ~450³/ha). A characteristic feature of the r50-sites is the occurrence of trees with 80 cm bhd or more with reduced vitality. The timber trunk) qualities of r-sites differ only slightly from managed stands. In the r50-sites the dead wood volume (>142 m³/ha) is much higher than in the w-sites (max. 34 m³/ha). Many different features of dead wood occur only fragmentary within w-sites. About 40 % of the dead wood objects have no "dead wood neighbour" (r50-sites: <2 %), and the light distribution is much less diverse. In >100 years unmanaged r-sites ~12 different types of special structures and 200 single special structures occur per ha. 19 out of 20 special structures are significantly less frequent in w-sites; 11 special structures are specifically valuable to be used as naturalness indicators.Vegetation: In the herb layer occur higher coverage values, more (light-indicating) species, but only few species indicating ancient forests and a higher diversity index value. In w-sites, the occurrence of e. g. Calamagrostis epigeios, Impatiens parviflora and Rubus idaeus is supported. reduced. Threatened moss species are rare in w-sites compared to r-sites, since they mainly grow on laying dead wood, which is rare in forests in use, and on inclined/rough-barked stem bases. Ground beetles: The forestry use of lowland beech forests leads to less individuals and lower biomass of so-called mesophilous forest species. Furthermore, the number of flightless individuals is lower. As proper indicators for near-natural lowland beech forests, the three species Carabus glabratus, C. hortensis und Cychrus caraboides could be identified. Indicators: 29 structural indicators were identified and thresholds were given. But even in lowland beech forests managed in a conservation-friendly way, these target values for near-natural and natural forests are unlikely to be reached. Therefore, for w-sites special threshold values have been defined, which consider the inevitable difference between managed and natural forests.