The Chemistry and Chemical Ecology of Lepidopterans as Investigated in Brazil.

The interdisciplinary field of Chemical Ecology in Brazil is currently composed of groups that emerged through the pioneering studies of Keith Spalding Brown Jr. and José Tércio Barbosa Ferreira. Following Keith Brown 's steps, José Roberto Trigo continued investigating the role of plant natural products in mediating the association among insects and their host plants, mainly in the Order Lepidoptera. The role of pyrrolizidine alkaloids in those associations was investigated extensively by Brown and Trigo, and most of what is currently known on this subject is based on their studies. The present work acknowledges their contribution to the Brazilian chemical ecology field and on insect-plant communication studies mediated by different chemical compounds.

Health risk associated with the oral consumption of "Chiniy-tref", a traditional medicinal preparation used in Martinique (French West Indies): Qualitative and quantitative analyses of aristolochic acids contained therein.

"Chiniy-tref" (CT) is a traditional preparation used in folk medicine in Martinique Island (French West Indies) that is nowadays mainly taken orally to prevent or act against any "manifestation of evil". CT is easily prepared at home by macerating larvae of the endemic swallowtail Battus polydamas (ssp.) cebriones (Dalman, 1823), sometimes accompanied by a leaf of its host-plant Aristolochia trilobata L., in commercial rum. We have previously reported the detection of nephrotoxic and carcinogenic aristolochic acids (AAs) I and II in CT, leading the Regional Health Agency (ARS) of Martinique to issue an alert regarding the potential risks associated with its consumption in 2015. In order to complete the toxicity risk assessment for oral consumption of CT, a full qualitative analysis of AAs and their analogues (AAAs) was performed, as well as a quantitative determination of the major AAs, namely AAs I and II. The phytochemical profiling of AAAs present in CT, that also corresponds to that of B. polydamas cebriones larvae feeding on A. trilobata, has been established for the first time by ultra-high performance liquid chromatography/electrospray ionization quadrupole time of-flight tandem mass spectrometry. AAs I and II were quantified in a small panel of tinctures by using a validated UHPLC/UV method, allowing us to estimate the probable daily intakes of these toxins by CT consumers. The results proved the existence of a real risk of renal toxicity and carcinogenicity associated with the chronic oral consumption of CT in Martinique, and more generally of similar "snake bottles" throughout the Caribbean.

Characterization of the fungitoxic activity on Botrytis cinerea of the aristolochic acids I and II.

The fungitoxic effect of aristolochic acids I and II on mycelial growth and conidial germination of Botrytis cinerea was analysed. Aristolochic acid I had a higher effect on mycelial growth of B. cinerea than aristolochic acid II with IC50 value of 18·7 and 57·0 µg ml-1 , respectively. These compounds did not affect the conidia germination. Also, the effect of both compounds on DNA and plasmatic membrane integrity of B. cinerea was studied. Only aristolochic acid II was able to cause damage to the integrity of the plasmatic membrane. When the fungus was incubated with a mixture of these compounds, degradation of DNA was observed. Finally, biotransformation products were not detected in the culture broth when B. cinerea was incubated in the presence of the aristolochic acids. Studies of structural characteristics that increase the antifungal effect of compounds against B. cinerea will permit to design new molecules to control this phytopathogenic fungus. SIGNIFICANCE AND IMPACT OF THE STUDY: The fungitoxic effect on Botrytis cinerea of aristolochic acids I and II was characterized. The only structural difference among these compounds is a methoxy group at carbon 8. However, despite their structural similarity, the fungitoxic effect of aristolochic acid I was higher than the effect of aristolochic acid II. This result suggests that the methoxy group is important for the fungitoxic activity of these compounds on B. cinerea.

Detection of aristolochic acids I and II in "Chiniy-trèf", a traditional medicinal preparation containing caterpillars feeding on Aristolochia trilobata L. in Martinique, French West Indies.

"Chiniy-trèf" is a traditional medicinal preparation used in Martinique, French West Indies, for the prevention of all kinds of attempted poisoning and hex. It is produced by the maceration in alcohol (mostly rum) of larvae (caterpillars) of the butterfly Battus polydamas ssp. cebriones, feeding on the leaves of Aristolochia trilobata. Aristolochic acids I and II that are well-known nephrotoxic and carcinogenic substances were identified on two samples of "chiniy-trèfl" by chromatographic methods.

Temporal Variation of Aristolochia chilensis Aristolochic Acids during Spring.

In this communication, we report the springtime variation of the composition of aristolochic acids (AAs) in Aristolochia chilensis leaves and stems. The dominant AA in the leaves of all samples, which were collected between October and December, was AA-I (1), and its concentration varied between 212.6±3.8 and 145.6±1.2 mg/kg and decreased linearly. This decrease occurred in parallel with the increase in AA-Ia (5) concentration from 15.9±0.8 mg/kg at the beginning of October to 96.8±7.8 mg/kg in mid-December. Both acids are enzymatically related by methylation-demethylation reactions. Other AAs also showed important variations: AA-II (2) significantly increased in concentration, reaching a maximum in the first two weeks of November and subsequently decreasing in mid-December to approximately the October levels. The principal component in the AA mixture of the stems was also AA-I (1); similar to AA-II (2), its concentration increased beginning in October, peaked in the second week of November and subsequently decreased. The concentrations of AA-IIIa (6) and AA-IVa (7) in the leaves and stems varied throughout the study period, but no clear pattern was identified. Based on the variation of AAs in A. chilensis leaves and stems during the study period, the reduced contents of non-phenolic AAs and increased concentrations of phenolic AAs are likely associated with a decrease in this plant's toxicity during the spring.