Introdução: a cromoblastomicose (CBM) é uma micose subcutânea causada por fungos melanizados de vários gêneros. Alternativas para identificação das espécies causadoras de CBM utilizando Espectroscopia no Infravermelho com Transformada de Fourier (FTIR) foi avaliada somente para o gênero Exophiala. Os testes de suscetibilidade aos antifúngicos não foram realizados para algumas espécies de agentes da CBM, sendo importante essa determinação para auxiliar na escolha adequada do tratamento. Embora o conhecimento de que a melanina tenha ação contra radicais livres oxidantes, poucos estudos exploraram o tema e somente o gênero Fonsecaea foi avaliado, utilizando baixo número de isolados. Objetivo: propor identificação dos agentes da CBM por FTIR e avaliar o perfil das diferentes espécies na resposta aos antifúngicos. Além disso, quantificar a melanina e avaliar a influência da mesma na sobrevivência dos isolados após estresse oxidativo e na suscetibilidade aos antifúngicos. Métodos: foram utilizados até 78 isolados clínicos em cada experimento. Para FTIR, a aquisição dos espectros de 4000-650 nm foi realizada a partir de técnica proposta; O teste de suscetibilidade a seis antifúngicos e o ensaio de estresse oxidativo frente ao peróxido de hidrogênio (H2O2) foram realizados com os isolados pareados (com e sem inibidor de melanina - triciclazol); A melanina foi extraída pelo método álcali-ácida e quantificada por gravimetria. Resultados: o modelo proposto de identificação por FTIR teve alto coeficiente de determinação e baixo erro da linha de tendência e validação cruzada obtidas. Terbinafina foi o antifúngico que apresentou menores concentrações inibitórias mínimas (CIMs) contra gêneros Fonsecaea, Phialophora e duas espécies de Rhinocladiella. Já anfotericina B e itraconazol apresentaram as CIMs mais elevadas. Além disso, outras diferenças entre gêneros e espécies foram observadas; gênero Rhinocladiella, quando a melanina não está inibida, mostrou maior sensibilidade ao estresse causado por H2O2 do que Phialophora e Fonsecaea, enquanto que os dois últimos foram os mais resistentes ao estresse oxidativo. Ao passo de que a quantidade de melanina extraída de Rhinocladiella foi significativamente menor do que Phialophora e Fonsecaea. Conclusão: Foi proposto um modelo que apresentou alta capacidade de classificar as 13 espécies de agentes causadores de CBM. A identificação e o teste de suscetibilidade aos antifúngicos nas práticas clínicas são importantes para auxiliar no tratamento. Além disso, os estudos que avaliaram a relação com melanina, tanto quantificação, estresse oxidativo e relação com antifúngicos comprovam, em agentes da CBM, o fator protetor da melanina frente a agressões externas sofridas por determinados gêneros e espécies, agressões estas relacionadas ao ambiente e ao hospedeiro. / Background: chromoblastomycosis (CBM) is a subcutaneous mycosis caused by melanized fungi of several genera. Alternative for species identification using Fourier Transform Infrared Spectroscopy (FTIR) was evaluated only for the genus Exophiala. The testing of antifungal susceptibity was not evaluated for some species of CBM agents, and this determination is important to aid in the appropriate choice of treatment. Although the knowledge that melanin has an action against oxidative radicals, few studies have explored the subject and only the genus Fonsecaea has been evaluated, using a low number of isolates. Objective: to propose identification of the CBM agents by FTIR and to evaluate the profile of the different species in the response to antifungals. Furthermore, to quantify the melanin and to evaluate its influence in the survival of the isolates after oxidative stress and the susceptibility to the antifungals. Methods: up to 78 clinical isolates were used in each experiment. For FTIR, the acquisition of the 4000-650 nm spectra was performed using a proposed technique; The susceptibility test using six antifungal agents and the oxidative stress test using hydrogen peroxide (H2O2) were performed with the isolates in paired form (with and without melanin inhibitor - tricyclazole); The melanin was extracted by the alkali-acid method and quantified by gravimetry. Results: the proposed FTIR identification model had a high coefficient of determination and a low error of the trend line and cross-validation obtained. Terbinafine was the antifungal that presented smaller minimum inhibitory concentrations (MICs) against genera Fonsecaea, Phialophora and two species of Rhinocladiella. While amphotericin B and itraconazole had the highest MICs. In addition, other differences between genera and species were observed; Genus Rhinocladiella, when melanin is not inhibited, showed greater sensitivity to stress caused by H2O2 than Phialophora and Fonsecaea, while the last two genera were the most resistant to oxidative stress. Moreover, the amount of melanin extracted from Rhinocladiella was significantly lower than Phialophora and Fonsecaea. Conclusion: a model that presented high capacity to classify the 13 species of CBM agents was proposed. Identification and testing of antifungal susceptibility in clinical settings are important to aid in treatment. Furthermore, the studies that evaluated the relationship with melanin, both quantification, oxidative stress and relation with antifungals demonstrate, in CBM agents, the protective factor of melanin against external aggressions of environmental and host suffered by certain genera and species.

Chionochloa, a genus of snow tussock grasses native to New Zealand, exhibits pronounced mast seeding. Chionochloa suffers very high levels of pre-dispersal flower and seed predation by three main insects: Eucalyptodiplosis chionochloae, a cecidomyiid midge, which is formally described here; Megacraspedus calamogonus, a gelechiid moth and Diplotoxa similis, a chloropid fly. Seven species of parasitoids that attack these seed predators were discovered. Four species parasitize M. calamogonus (one tachinid fly and three hymenopteran wasps), one parasitizes D. similis (Hymenoptera: Eulophidae) and two parasitize E. chionochloae, (a pteromalid wasp Gastrancistrus sp. and a platygastrid wasp Zelostemma chionochloae, which is given a formal description here). The abundance, predation levels by each of the insect species, and interactions between all the organisms in the community were studied across three elevations at Mount Hutt over three summer seasons. M. calamogonus was most abundant at 450 m altitude during all three seasons. D. similis was most common at 1070 m altitude, while its predation levels peaked in low flowering seasons and decreased in high seasons. E. chionochloae was abundant in all three altitudes and increased its predation levels with increasing flowering intensity. E. chionochloae was confirmed to use prolonged diapause of at least three years. Prolonged diapause was also confirmed in its two parasitoids. Chionochloa plants were manipulated with various treatments to test the effect on diapause in E. chionochloae and its two parasitoids. Treatments included plant warming, root pruning, gibberellic acid sprayed on the plants and combinations of these treatments. All three insects changed their emergence in response to some treatments and therefore it was suggested that combined with risk-spreading diapause, they may use some predicting to emerge from prolonged diapause. E. chionochloae control their diapause following some of the cues that Chionochloa use for flowering, while Z. chionochloae and Gastrancistrus in some cases follow their host’s cues and in others use similar cues as Chionochloa plants. Emergence or diapause predictions differed across elevations and plant species in all three insect seed/flower predators. E. chionochloae had female-biased sex ratios in different populations even after prolonged diapause. There was week evidence that both parasitoid species are female-biased in the first emergence year and male-biased after more than one year in diapause. Therefore it was suggested that diapause is not more costly for females of E. chionochloae and its parasitoid than for males. Females of all three species were not found to be better predictors (i.e, more likely to respond to treatments by not entering extended diapause) than males. The complex interactions of all the organisms in this web are thought to be sensitive to climate, and it was suggested that the global climate change may alter this sensitive system.

Chionochloa, a genus of snow tussock grasses native to New Zealand, exhibits pronounced mast seeding. Chionochloa suffers very high levels of pre-dispersal flower and seed predation by three main insects: Eucalyptodiplosis chionochloae, a cecidomyiid midge, which is formally described here; Megacraspedus calamogonus, a gelechiid moth and Diplotoxa similis, a chloropid fly. Seven species of parasitoids that attack these seed predators were discovered. Four species parasitize M. calamogonus (one tachinid fly and three hymenopteran wasps), one parasitizes D. similis (Hymenoptera: Eulophidae) and two parasitize E. chionochloae, (a pteromalid wasp Gastrancistrus sp. and a platygastrid wasp Zelostemma chionochloae, which is given a formal description here). The abundance, predation levels by each of the insect species, and interactions between all the organisms in the community were studied across three elevations at Mount Hutt over three summer seasons. M. calamogonus was most abundant at 450 m altitude during all three seasons. D. similis was most common at 1070 m altitude, while its predation levels peaked in low flowering seasons and decreased in high seasons. E. chionochloae was abundant in all three altitudes and increased its predation levels with increasing flowering intensity. E. chionochloae was confirmed to use prolonged diapause of at least three years. Prolonged diapause was also confirmed in its two parasitoids. Chionochloa plants were manipulated with various treatments to test the effect on diapause in E. chionochloae and its two parasitoids. Treatments included plant warming, root pruning, gibberellic acid sprayed on the plants and combinations of these treatments. All three insects changed their emergence in response to some treatments and therefore it was suggested that combined with risk-spreading diapause, they may use some predicting to emerge from prolonged diapause. E. chionochloae control their diapause following some of the cues that Chionochloa use for flowering, while Z. chionochloae and Gastrancistrus in some cases follow their host’s cues and in others use similar cues as Chionochloa plants. Emergence or diapause predictions differed across elevations and plant species in all three insect seed/flower predators. E. chionochloae had female-biased sex ratios in different populations even after prolonged diapause. There was week evidence that both parasitoid species are female-biased in the first emergence year and male-biased after more than one year in diapause. Therefore it was suggested that diapause is not more costly for females of E. chionochloae and its parasitoid than for males. Females of all three species were not found to be better predictors (i.e, more likely to respond to treatments by not entering extended diapause) than males. The complex interactions of all the organisms in this web are thought to be sensitive to climate, and it was suggested that the global climate change may alter this sensitive system.