Total Pyrrolizidine Alkaloids Concentration Research Articles

Profiling of pyrrolizidine alkaloids using a retronecine-based untargeted metabolomics approach coupled to the quantitation of the retronecine-core in medicinal plants using UHPLC-QTOF

Pyrrolizidine alkaloids (PA) are secondary metabolites of high toxicological relevance. Several PA quantitative methodologies were developed based on a limited number of certified standards, including time consuming solid phase extraction (SPE) purification steps. Herein, we shed light on the variability of PA in herbal extracts and propose a quantification methodology based on ultra-high-performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS) for the evaluation of the total PA content as retronecine-equivalents (RE) directly from crude matrices. Particularly in the focus of the investigation were Alkanna spp. (Boraginaceae), which possess a wide range of pharmaceutical properties. In addition, a comparative PA screening of crude and SPE enriched extracts was performed and PA-containing plants from Fabaceae and Compositae families were included to demonstrate universal applicability. In total, 105 PA were identified using HRMSe experiments, specific MS/MS fragmentation PA patterns, a customized in-house library and literature data. Among them, 18 glycosidic PA derivatives were reported for the first time in literature. Using a hierarchical clustering approach, PA distribution in herbal extracts was shown to be family-dependent and significantly different among species. This was further supported by the results of the total PA concentrations, obtained using a retronecine/heliotridine/internal standard-based targeted UHPLC-HRMS quantification method, which varied from 8.64 ± 0.08–3096.28 ± 273.72 μg RE/g extract dry weight in shoots extracts of Alkanna spp. and leaves extracts of Crotalaria retusa L. respectively. Worth mentioning is that the procedure allowed to quantify PA in Alkanna spp. If the procedure based on 35 specific PA recommended by European regulations had been used, results would have been equal to zero for the four species since none were observed in Alkanna spp. Finally, by combining the RE results with the corresponding dereplication results, a customized correction factor for each extract (ranging from 2.12 to 2.48) was assessed leading to a more accurate estimate of the PA content regardless of the molecular weight of each PA. The present methodology will facilitate PA quantification directly from crude extracts and avoid the underestimation the real PA content due to limited availabilty of authentic reference compounds in botanical extracts used in phytomedicines or food supplements/cosmetics.

Seasonal variation in defence compounds: A case study on pyrrolizidine alkaloids of clones of Jacobaea vulgaris, Jacobaea aquatica and their hybrids

Concentration of plant secondary metabolites (SMs) show seasonal variations. However, it is still not well understood how these abiotic and biotic factors influence the seasonal variations of SMs. In addition, it is of interest to know if and how SMs are reallocated to the different plant organs, in particular whether SMs are reallocated to the remaining tissues when biomass is lost, e.g., during winter. Here we used Jacobaea vulgaris, Jacobaea aquatica, two F1 and four F2 hybrids that differed in their pyrrolizidine alkaloids (PAs) bouquet as a study system. A series of clones of these genotypes were investigated during their vegetative stage spanning 14 months in a semi-natural environment. We found that the total PA concentration in roots and shoots showed a gradual increase until the spring of the second year, whereafter it dropped substantially in shoots. The variation in PA composition due to seasonal changes was significant but relatively small. Senecionine-like PAs were the dominant PAs in roots, while jacobine-/erucifoline-like PAs were dominant in shoots. The variation of PA concentration was significantly correlated with temperature, day length, and plant age. A correlation analysis showed that PAs were not reallocated when biomass was lost in winter. Overall, our study showed that PA composition of each genotype changed over seasons in a different manner but seasonal variation did not overrule the differences in PA composition among genotypes.

Retrospective HRMS Screening and Dedicated Target Analysis Reveal a Wide Exposure to Pyrrolizidine Alkaloids in Small Streams.

Pyrrolizidine alkaloids (PAs) are found to be toxic pollutants emitted into the environment by numerous plant species, resulting in contamination. In this article, we investigate the occurrence of PAs in the aquatic environment of small Swiss streams combining two different approaches. Pyrrolizidine alkaloids (PAs) are toxic secondary metabolites produced by numerous plant species. Although they were classified as persistent and mobile and found to be emitted into the environment, their occurrence in surface waters is largely unknown. Therefore, we performed a retrospective data analysis of two extensive HRMS campaigns each covering five small streams in Switzerland over the growing season. All sites were contaminated with up to 12 individual PAs and temporal detection frequencies between 36 and 87%. Individual PAs were in the low ng/L range, but rain-induced maximal total PA concentrations reached almost 100 ng/L in late spring and summer. Through PA patterns in water and plants, several species were tentatively identified as the source of contamination, with Senecio spp. and Echium vulgare being the most important. Additionally, two streams were monitored, and PAs were quantified with a newly developed, faster, and more sensitive LC-MS/MS method to distinguish different plant-based and indirect human PA sources. A distinctly different PA fingerprint in aqueous plant extracts pointed to invasive Senecio inaequidens as the main source of the surface water contamination at these sites. Results indicate that PA loads may increase if invasive species are sufficiently abundant.

Evolution of Increased Photosynthetic Capacity and Its Underlying Traits in Invasive Jacobaea vulgaris.

The evolution of increased competitive ability (EICA) hypothesis and the shifting defense hypothesis (SDH) predict that evolutionary changes occur in a suite of traits related to defense and growth in invasive plant species as result of the absence of specialist herbivores. We tested how this suite of traits changed due to the absence of specialist herbivores in multiple invasive regions that differ in climatic conditions with native and invasive Jacobaea vulgaris in a controlled environment. We hypothesized that invasive J. vulgaris in all invasive regions have i) a higher plant growth and underlying traits, such as photosynthetic capacity, ii) lower regrowth-related traits, such as carbohydrate storage, and iii) an increased plant qualitative defense, such as pyrrolizidine alkaloids (PAs). Our results show that invasive J. vulgaris genotypes have evolved a higher photosynthetic rate and total PA concentration but a lower investment in root carbohydrates, which supports the SDH hypothesis. All the traits changed consistently and significantly in the same direction in all four invasive regions, indicative of a parallel evolution. Climatic and soil variables did differ between ranges but explained only a very small part of the variation in trait values. The latter suggests that climate and soil changes were not the main selective forces on these traits.

Occurrence of Nine Pyrrolizidine Alkaloids in Senecio vulgaris L. Depending on Developmental Stage and Season.

The contamination of phytopharmaceuticals and herbal teas with toxic plants is an increasing problem. Senecio vulgaris L. is a particularly noxious weed in agricultural and horticultural crops due to its content of toxic pyrrolizidine alkaloids (PAs). Since some of these compounds are carcinogenic, the distribution of this plant should be monitored. The amount of PAs in S. vulgaris is affected by various factors. Therefore, we investigated the occurrence of PAs depending on the developmental stage and season. A systematic study using field-plot experiments (four seasons, five developmental stages of the plants: S1 to S5) was performed and the PA concentration was determined via LC-MS/MS analysis. The total amount of PAs in the plant increased with the plant development, however, the total PA concentrations in µg/g dry matter remained nearly unchanged, whilst trends for specific PAs were observed. The concentrations of PA-N-oxides (PANOs) were much higher than that of tertiary PAs. Maximal amounts of the PA total were 54.16 ± 4.38 mg/plant (spring, S5). The total amount of PAs increased strongly until later developmental stages. Therefore, even small numbers of S. vulgaris may become sufficient for relevant contaminations set out by the maximal permitted daily intake levels recommended by the European Food Safety Authority (EFSA).

Methyl Jasmonate Changes the Composition and Distribution Rather than the Concentration of Defence Compounds: a Study on Pyrrolizidine Alkaloids

In this study we investigated the effect of methyl jasmonate (MeJA) application on pyrrolizidine alkaloid (PA) concentration and composition of two closely related Jacobaea species. In addition, we examined whether MeJA application affected herbivory of the polyphagous leaf feeding herbivore Spodoptera exigua. A range of concentrations of MeJA was added to the medium of Jacobaea vulgaris and J. aquatica tissue culture plants grown under axenic conditions. PA concentrations were measured in roots and shoots using LC-MS/MS. In neither species MeJA application did affect the total PA concentration at the whole plant level. In J. vulgaris the total PA concentration decreased in roots but increased in shoots. In J. aquatica a similar non-significant trend was observed. In both Jacobaea species MeJA application induced a strong shift from senecionine- to erucifoline-like PAs, while the jacobine- and otosenine-like PAs remained largely unaffected. The results show that MeJA application does not necessarily elicits de novo synthesis, but rather leads to PA conversion combined with reallocation of certain PAs from roots to shoots. S. exigua preferred feeding on control leaves of J. aquatica over MeJA treated leaves, while for J. vulgaris both the control and MeJA treated leaves were hardly eaten. This suggests that the MeJA-induced increase of erucifoline-like PAs can play a role in resistance of J. aquatica to S. exigua. In J. vulgaris resistance to S. exigua may already be high due to the presence of jacobine-like PAs or other resistance factors.

Analytical strategies for the determination of pyrrolizidine alkaloids in plant based food and examination of the transfer rate during the infusion process

Two sample preparation methods were developed (graphitised carbon and C18 solid phase extraction clean-up) and validated in house using liquid chromatography and tandem mass spectrometry (MS/MS) for the determination of 30 pyrrolizidine alkaloids (PAs) in salads, herbs, tea, herbals teas as well as tea infusion and ice-tea beverages. Total PAs concentration of samples purchased on the Belgian market varied greatly with matrix type ranging from <LOD to 187151 ng/g for dry samples, while for infusions and ice-tea beverages the highest PAs concentration was 2106 ng/mL. Surprisingly high PAs concentrations were detected in herbs and spices mixes. The infusion study indicated a transfer rate between 16 and 28% (except for monocrotaline) which highlights the overestimation of PAs concentrations in infusions when derived from a dilution factor (transfer rate of 100%) to the measured concentrations of the dry tea sample.

Pyrrolizidine alkaloid variation in Senecio vulgaris populations from native and invasive ranges.

Biological invasion is regarded as one of the greatest environmental problems facilitated by globalization. Some hypotheses about the invasive mechanisms of alien invasive plants consider the plant–herbivore interaction and the role of plant defense in this interaction. For example, the “Shift Defense Hypothesis” (SDH) argues that introduced plants evolve higher levels of qualitative defense chemicals and decreased levels of quantitative defense, as they are released of the selective pressures from specialist herbivores but still face attack from generalists. Common groundsel (Senecio vulgaris), originating from Europe, is a cosmopolitan invasive plant in temperate regions. As in other Senecio species, S. vulgaris contains pyrrolizidine alkaloids (PAs) as characteristic qualitative defense compounds. In this study, S. vulgaris plants originating from native and invasive ranges (Europe and China, respectively) were grown under identical conditions and harvested upon flowering. PA composition and concentration in shoot and root samples were determined using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS). We investigated the differences between native and invasive S. vulgaris populations with regard to quantitative and qualitative variation of PAs. We identified 20 PAs, among which senecionine, senecionine N-oxide, integerrimine N-oxide and seneciphylline N-oxide were dominant in the roots. In the shoots, in addition to the 4 PAs dominant in roots, retrorsine N-oxide, spartioidine N-oxide and 2 non-identified PAs were also prevalent. The roots possessed a lower PA diversity but a higher total PA concentration than the shoots. Most individual PAs as well as the total PA concentration were strongly positively correlated between the roots and shoots. Both native and invasive S. vulgaris populations shared the pattern described above. However, there was a slight trend indicating lower PA diversity and lower total PA concentration in invasive S. vulgaris populations than native populations, which is not consistent with the prediction of SDH.

Phytochemical investigations and food-choice experiments with two mollusc species in three central European Senecio L. (Asteraceae, Senecioneae) species and their hybrids

Differences in herbivore resistance between parental and hybrid taxa influence the trajectories of hybrid swarms and may lead to the maintenance or breakdown of species boundaries. In the present study, phytochemical patterns in terms of pyrrolizidine alkaloids (PAs) and tannins in three central European representatives of the Senecio nemorensis syngameon (i.e., S. germanicus, S. hercynicus, S. ovatus) were inferred and the selective consumption among the pure species and among the pure species and their reciprocal hybrids was studied in food-choice (‘cafeteria’) experiments using two mollusc species. The results show significant qualitative and quantitative differences in chemical composition among the three plant species. Artificial F 1 hybrids of the two investigated plant systems S. germanicus–S. ovatus and S. hercynicus–S. ovatus mainly expressed an additive PA pattern compared to the parental taxa. PA concentration was highest in S. hercynicus, intermediate in S. ovatus and lowest in S. germanicus, while tannin concentration was lower in S. hercynicus compared to S. ovatus and S. germanicus. Food-choice experiments revealed significant plant species-specific preferences exhibited by the two mollusc species (the slug Arion fuscus and the snail Trochulus striolatus danubialis). Consistent negative correlations between the total PA concentration and the consumed leaf area were only observed for the snail, but not for the slug species. When reciprocal hybrid taxa were added to the food-choice designs, the plant taxon-specific differences in consumption disappeared. Additionally, no significant effect of tannin concentration on strength of herbivory was observed. The present results of food-choice experiments in three representatives of the S. nemorensis syngameon, therefore, demonstrate that PAs rather than tannins influence taxon-specific herbivory by molluscs. However, they also suggest that trajectories of mixed stands and hybrid swarms in the wild could be more pronouncedly determined by the local composition of mollusc communities, the frequency of intermediate chemotypes (hybrids), and other factors (e.g., morphology, other biotic and abiotic conditions).

Survey of pyrrolizidine alkaloids in seven varieties of Lappula squarrosa: An alternative source of heart-healthy vegetable oil.

Growing demand for heart-healthy omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), is putting stress on wild fish stocks. There is now a compelling need for new and novel sources of non-traditional seed oils containing high stearidonic acid (SDA), a precursor of EPA and DHA, to reduce this demand. The seed oil of Lappula squarrosa is one of the richest sources of SDA, however, the plant has been found to contain toxic pyrrolizidine alkaloids (PAs). In this study, the PA concentrations of seven varieties (A-G) of Lappula squarrosa were analysed to determine the most suitable varieties for commercial seed oil production. Whilst the clean-up procedure for the PAs in the roots, flowers and leaves was on diatomaceous earth columns and finally analysed with GC-EI-MS, that of the seeds was through SCX-SPE and a more sensitive HPLC-ESI-MS/MS sum parameter method was used in the analysis. Altogether six PAs (supinine, amabiline, intermedine, lycopsamine and 3'-acetylintermedine) including one unknown retronecine-type PA were identified with variety C recording the lowest total PA concentration (4.64 mg seneciphylline equivalents (SE)/g dry weight (d.w.)). Besides, the total PA concentrations in the seeds of Lappula squarrosa varieties ranged between 2.88 μg PA/g and 10.36 μg PA/g d.w. Based solely on overall PA concentrations and PA distribution, variety D (5.95 mg SE/g d.w.) was found to be a potential candidate for commercial seed oil cultivation.

Interactive effects of above- and belowground herbivory and plant competition on plant growth and defence

Competition and herbivory are two major factors that can influence plant growth and plant defence. Although these two factors are often studied separately, they do not operate independently. We examined how aboveground herbivory by beet armyworm larvae (Spodoptera exigua) and belowground herbivory by wireworms (Agriotes lineatus) influenced competition between the plant species Jacobaea vulgaris and Leucanthemum vulgare exposed to three competition levels (no, intra-, and interspecific competition). In addition, we studied the effects of herbivory and competition on pyrrolizidine alkaloid (PA) concentrations in leaves of J. vulgaris. For J. vulgaris, aboveground herbivory significantly reduced shoot biomass while belowground herbivory increased root biomass. Biomass of L. vulgare was not affected by herbivory. Competition caused a reduction in biomass for both plant species, but herbivory did not affect the outcome of the competition. However, competition significantly influenced the amount of leaf damage experienced by the plants. A L. vulgare plant had significantly less damage from aboveground herbivores when grown together with J. vulgaris than when grown alone or in intraspecific competition, while a J. vulgaris plant experienced lowest damage in conditions of intraspecific competition. The total PA concentration in J. vulgaris leaves was highest for plants exposed to interspecific competition. Root herbivory caused an increase in the relative concentration of N-oxides, the less toxic form of PAs, in leaves of plants that were grown without competition, but a decrease in plants exposed to competition. Our study shows that competition and herbivory but also the type of competition and whether herbivory occurs above- or belowground, all influence plant performance. However, overall, there was no evidence that herbivory affects plant–plant competition.

Hepatotoxic pyrrolizidine alkaloids in Emilia sonchifolia from Taiwan

Emilia sonchifolia (L.) DC. (Lilac tasselflower, Zi Bei Cao) is used in Asian countries for food or medicinal purposes. We have investigated the hepatotoxic pyrrolizidine alkaloids (PAs) contained in E. sonchifolia. The objective was to determine profile and quantity of the PAs in E. sonchifolia samples that were collected from several towns in Taiwan. Pyrrolizidine alkaloids were isolated and purified from the aqueous acid extracts by use of strong cation exchange-solid phase extraction (SCX-SPE). Alkaloid extracts were analyzed using gas–chromatography mass spectrometry (GC–MS). Eleven PAs of two types were identified in E. sonchifolia (by type): (1) retronecine bases, senecionine, seneciphylline and integerrimine; (2) otonecine bases, senkirkine, otosenine, neosenkirkine, petasitenine, acetylsenkirkine, desacetyldoronine, acetylpetasitenine, and doronine. Quantification of the individual PAs was determined by six-point linear regression curves in the range of 25–400μg/mL of senkirkine and senecionine. The PAs were found in all plant organs of E. sonchifolia. Senkirkine was the major PA identified with concentrations up to 53.8μg/g of dry herb. The average total PA concentration varied from a low of 33.3μg/g to a high of 93.9μg/g dry herb. This study will provide important information to public consumers regarding the health risk of PA-containing E. sonchifolia.

The Relationship between Structurally Different Pyrrolizidine Alkaloids and Western Flower Thrips Resistance in F2 Hybrids of Jacobaea vulgaris and Jacobaea aquatica

Segregating plant hybrids often have more ecological and molecular variability compared to parental species, and are therefore useful for studying relationships between different traits, and the adaptive significance of trait variation. Hybrid systems have been used to study the relationship between the expression of plant defense compounds and herbivore susceptibility. We conducted a western flower thrips (WFT) bioassay using a hybrid family and investigated the relationship between WFT resistance and pyrrolizidine alkaloid (PA) variation. The hybrid family consisted of two parental (Jacobaea vulgaris and Jacobaea aquatica) genotypes, two F1 genotypes, and 94 F2 hybrid lines. The J. aquatica genotype was more susceptible to thrips attack than the J. vulgaris genotype, the two F1 hybrids were as susceptible as J. aquatica, and susceptibility to WFT differed among F2 hybrid lines: 69 F2 lines were equally susceptible compared to J. aquatica, 10 F2 lines were more susceptible than J. aquatica and 15 F2 lines were as resistant as J. vulgaris or were intermediate to the two parental genotypes. Among 37 individual PAs that were derived from four structural groups (senecionine-, jacobine-, erucifoline- and otosenine-like PAs), the N-oxides of jacobine, jaconine, and jacoline were negatively correlated with feeding damage caused by WFT, and the tertiary amines of jacobine, jaconine, jacoline, and other PAs did not relate to feeding damage. Total PA concentration was negatively correlated with feeding damage. Among the four PA groups, only the total concentration of the jacobine-like PAs was negatively correlated with feeding damage. Multiple regression tests suggested that jacobine-like PAs play a greater role in WFT resistance than PAs from other structural groups. We found no evidence for synergistic effects of different PAs on WFT resistance. The relationship between PA variation and WFT feeding damage in the Jacobaea hybrids suggests a role for PAs in resistance to generalist insects.Electronic supplementary materialThe online version of this article (doi:10.1007/s10886-011-0021-6) contains supplementary material, which is available to authorized users.

Arbuscular mycorrhizal colonization, plant chemistry, and aboveground herbivory on Senecio jacobaea

Arbuscular mycorrhizal fungi (AMF) can affect insect herbivores by changing plant growth and chemistry. However, many factors can influence the symbiotic relationship between plant and fungus, potentially obscuring experimental treatments and ecosystem impacts. In a field experiment, we assessed AMF colonization levels of individual ragwort ( Senecio jacobaea) plants growing in grassland plots that were originally sown with 15 or 4 plant species, or were unsown. We measured the concentrations of carbon, nitrogen and pyrrolizidine alkaloids (PAs), and assessed the presence of aboveground insect herbivores on the sampled plants. Total AMF colonization and colonization by arbuscules was lower in plots sown with 15 species than in plots sown with 4 species and unsown plots. AMF colonization was positively related to the cover of oxeye daisy ( Leucanthemum vulgare) and a positive relationship between colonization by arbuscules and the occurrence of a specialist seed-feeding fly ( Pegohylemyia seneciella) was found. The occurrence of stem-boring, leaf-mining and sap-sucking insects was not affected by AMF colonization. Total PA concentrations were negatively related to colonization levels by vesicles, but did not differ among the sowing treatments. No single factor explained the observed differences in AMF colonization among the sowing treatments or insect herbivore occurrence on S. jacobaea. However, correlations across the treatments suggest that some of the variation was due to the abundance of one plant species, which is known to stimulate AMF colonization of neighbouring plants, while AMF colonization was related to the occurrence of a specialist insect herbivore. Our results thus illustrate that in natural systems, the ecosystem impact of AMF through their influence on the occurrence of specialist insects can be recognised, but they also highlight the confounding effect of neighbouring plant species identity. Hence, our results emphasise the importance of field studies to elucidate interactions between AMF and organisms of different trophic levels.

Species by Environment Interactions Affect Pyrrolizidine Alkaloid Expression in Senecio jacobaea, Senecio aquaticus, and Their Hybrids

We examined the effects of water and nutrient availability on the expression of the defense pyrrolizidine alkaloids (PAs) in Senecio jacobaea and S. aquaticus. Senecio jacobaea, and S. aquaticus are adapted to different natural habitats, characterized by differing abiotic conditions and different selection pressures from natural enemies. We tested if PA concentration and diversity are plastic over a range of water and nutrient treatments, and also whether such plasticity is dependent on plant species. We also tested the hypothesis that hybridization may contribute to PA diversity within plants, by comparing PA expression in parental species to that in artificially generated F1 hybrids, and also in later generation natural hybrids between S. jacobaea and S. aquaticus. We showed that total PA concentration in roots and shoots is not dependent on species, but that species determines the pattern of PA diversification. Pyrrolizidine alkaloid diversity and concentration are both dependent on environmental factors. Hybrids produce a putatively novel PA, and this PA is conserved in natural hybrids, that are backcrossed to S. jacobaea. Natural hybrids that are backcrossed several times to S. jacobaea are with regard to PA diversity significantly different from S. jacobaea but not from S. aquaticus, while F1 hybrids are in all cases more similar to S. jacobaea. These results collectively suggest that PA diversity is under the influence of natural selection.

Chemotype of Senecio jacobaea affects damage by pathogens and insect herbivores in the field

The evolution of the diversity of plant secondary metabolites is still poorly understood. To determine whether natural enemies could exert selection on plant secondary chemistry, pathogen infestation and invertebrate herbivory were measured on 10 genotypes of Senecio jacobaea (Tansy Ragwort) at two experimental field sites during a 2-year period. The genotypes represented two chemotypes based on the presence of the pyrrolizidine alkaloids (PA) jacobine and erucifoline. At one site, Heteren, mainly generalist herbivores were present. Here, damage was limited and did not differ among genotypes or chemotypes. At the other site, Meijendel, several specialists attacked the plants. Damage increased during the year, with a peak in July when most damage was caused by the specialist moth Tyria jacobaeae. At this peak there was no difference in damage among chemotypes. In the months prior to T. jacobaeae damage, chemotypes with jacobine were more severely attacked by specialists than the chemotypes without jacobine. Total damage during that period was positively correlated with both total PA concentration and jacobine concentration. Probably plant vigor also played a role in host preference since damage per individual plant was positively correlated with plant size. Our results suggest that total PA concentration and specifically jacobine had a positive effect on specialist feeding, indicating ecological costs involved in the production of PAs. Ecological costs related to plant secondary compounds could explain why not all individuals produce high levels of these compounds. In addition, differences in specialist herbivore pressures among sites may contribute to the variation in secondary metabolites among populations.

Chemical ecology of the cinnabar moth (Tyria jacobaeae) on a newly recorded host Senecio adonidifolius

Abstract The cinnabar moth ( Tyria jacobaeae , Arctiidae) normally feeds on Senecio jacobaea in the field. For the first time, naturally occurring populations of T. jacobaeae have been found thriving on Senecio adonidifolius , even though the moth's preferred host, S. jacobaea , is available within 50–400 m. In the laboratory, the cinnabar moth has been shown to feed on and develop on S. adonidifolius despite its different leaf morphology, pyrrolizidine alkaloid (PA) profile and a large taxonomic distance to S. jacobaea . Here I examined whether T. jacobaeae has adapted to this new host in the field using adult oviposition behavior and plant acquired defense chemistry in pupae as criteria. Choice tests indicated local adaptation to this newly recorded host. T. jacobaeae reared on S. adonidifolius hosts laid more egg batches and total eggs on it than T. jacobaeae from S. jacobaea . The egg batches were smaller on S. adonidifolius possibly due to highly pinnate thread-like structure of its leaves. The bouquet of plant acquired PAs and the insect metabolized callimorphine in pupae differed widely between pupae collected from the two hosts. T. jacobaeae pupae taken from S. adonidifolius hosts contained more of the insect metabolized callimorphine than pupae taken from S. jacobaea hosts, but they did not differ in total PA concentration. Pupae taken from S. jacobaea hosts contained more unmetabolized plant PA's than pupae from S. adonidifolius hosts. Additionally, 10% of T. jacobaeae larvae taken from S. adonidifolius in Biausse were infested with Carcelia dubia , a parasitic and rare tachinid fly that typically attacks arctiid moths.

Aspectos epidemiológicos da seneciose na região sul do Rio Grande do Sul

Seneciose é a principal intoxicação por planta no sul do Rio Grande do Sul, Brasil. Neste trabalho apresentam-se os dados de 24 surtos de seneciose em bovinos e de um em eqüinos, diagnosticados pelo Laboratório Regional de Diagnóstico (LRD) da Universidade Federal de Pelotas, no período de 1998 a 2000. Adicionalmente são avaliados os dados epidemiológicos de 54 surtos ocorridos no período 1978-1997. Os dados epidemiológicos e clínicos do período 1998-2000 foram coletados em visitas às propriedades onde estavam ocorrendo os surtos. Em 11 (45,83%) surtos os animais tinham até 3 anos de idade e, em 13 (54,16%), 3 anos ou mais. Quanto ao sexo, 9 (37,5%) surtos atingiram fêmeas e 15 (62,5%) afetaram machos. Em relação à época de ocorrência, 10 (41,66%) surtos ocorreram na primavera, 4 (16,66%) no verão, 5 (20,83%) no outono, e 5 (20,83%) no inverno. A morbidade foi estimada em 4,92% e a letalidade em 95,59%. As espécies de Senecio que predominaram nas propriedades visitadas foram S. brasiliensis em 12 (57,14%), S. selloi em 10 (47,61%), S. oxyphyllus em 6 (28,57%), S. heterotrichius em 3 (14,28%), e S. leptolobus em 1 (4,76%) propriedade. Os sinais clínicos mais freqüentes foram emagrecimento progressivo, incoordenação, diarréia, tenesmo, prolapso retal e agressividade. O curso clínico da intoxicação, levando em consideração somente os animais necropsiados ou os observados durante as visitas às fazendas, foi de 24 a 96 horas em 4 (16,66%) surtos, de 4 a 7 dias em 7 (29,16%), de 1 a 2 semanas em 4 (16,66%), de 2 a 3 semanas em 2 (8,33%), de 1 a 2 meses em 2 (8,33%), e de 2 a 3 meses em 1 (4,16%) surto. Em quatro (16,66%) surtos o período de manifestação clínica não foi observado. Nos surtos em que o curso clínico foi mais prolongado, alguns animais apresentaram fotossensibilização. Os achados de necropsia mais comuns foram fígado aumentado de tamanho e endurecido, vesícula biliar aumentada, edema no mesentério e abomaso e líquido nas cavidades. No fígado, a histopatologia revelou fibroplasia, megalocitose e proliferação de células dos ductos biliares e, no cérebro, degeneração da substância branca. Em amostras das diferentes espécies de Senecio, coletadas em diferentes épocas do ano, foi determinada a concentração de alcalóides pirrolizidínicos (APs) e seus N-óxidos através de cromatografia de camada delgada e espectrofotometria. Retrorsina foi encontrada em Senecio brasiliensis, S. heterotrichius, S. selloi e S. oxyphyllus. Além deste, S. brasiliensis e S. heterotrichius registraram mais um e dois alcalóides não identificados, respectivamente. A maior concentração de AP reduzido (AP livre + N-óxido) foi encontrada em S. brasiliensis no inverno (0,25%). Nessa mesma época quantidades decrescentes de AP reduzidos foram encontradas em S. heterotrichius (0,19%), S. oxyphyllus (0,03%) e S. selloi (0,03%). Em S. leptolobus não foram detectados alcalóides. Esses dados demonstraram que S. brasiliensis é a espécie mais importante como causa de seneciose no sul do Rio Grande do Sul. Adicionalmente, foram analisados dados de arquivo do LRD sobre 54 surtos de intoxicação por AP no período 1978-1997. Nesse período, 7 (12,96%) surtos afetaram bovinos menores de 3 anos, 39 (72,22%) afetaram bovinos de 3 anos ou mais, 3 (5,55%) afetaram bovinos de várias idades e, em 5 (9,25%) surtos a idade não foi informada. Trinta e nove (72,22%) surtos afetaram fêmeas, 7 (12,96%) afetaram machos, 3 (5,55%) afetaram ambos os sexos e, em 5 (9,25%), o sexo não foi informado. Vinte e três (42,59%) surtos ocorreram na primavera, 9 (16,66%) no verão, 9 (16,66%) no outono, e 13 (24,07%) no inverno. O maior número de surtos da intoxicação no período 1998-2000 (24 surtos em 3 anos) em relação ao período 1978-1997 (54 surtos em 20 anos) pode estar relacionado à diminuição em mais de 50% da população ovina no Estado.

Variation in pyrrolizidine alkaloid patterns of Senecio jacobaea

We studied the variation in pyrrolizidine alkaloid (PA) patterns of lab-grown vegetative plants of 11 European Senecio jacobaea populations. Plants were classified as jacobine, erucifoline, mixed or senecionine chemotypes based on presence and absence of the PAs jacobine or erucifoline. Due to the presence of jacobine, total PA concentration in jacobine chemotypes was higher than in erucifoline chemotypes. Both relative and absolute concentrations of individual PAs differed between half-sib and clonal families, which showed that variation in PA patterns had a genetic basis. Within most populations relative abundance of PAs varied considerably between individual plants. Most populations consisted either of the jacobine chemotype or of the erucifoline chemotype, sometimes in combination with mixed or senecionine chemotypes.

Nutrients decrease pyrrolizidine alkaloid concentrations in Senecio jacobaea

• Changes in the defence compounds pyrrolizidine alkaloids (PAs) in roots and shoots of Senecio jacobaea are reported in response to nutrient addition in order to investigate whether changes in concentration are adaptive. • PA concentrations were examined in leaves and roots of 40 vegetative ragwort plants, subjected to four nutrient treatments in a climate chamber study. Roots from 10 plants were subdivided into main root cortex, main root vascular cylinder, lateral roots and root tips and analysed for PA concentrations. • Increasing nutrients lead to a significant reduction in total PA concentration of both roots and shoots. All individual PAs except jacobine decreased in concentration. The total amount of PA produced in the whole plant was not influenced by nutrient supply. Root tips contained a three times lower concentration than the main and lateral roots. The concentrations in the main root cortex were five times higher than concentrations in the vascular cylinder. • Changes in biomass rather than changes in production rates can explain alterations in PA concentration of S. jacobaea in response to nutrients.