Determination of cocaine in some south american species of Erythroxylum using mass fragmentography

Thirteen South American species of Erythroxylum have been analyzed for their cocaine content. Cocaine was found only in E. coca Lam., E. novogranatense (Morris) Hieron. and E. novogranatense var. truxillense (Rusby) Machado. The amount of cocaine was determined by mass fragmentography using deuterium labelled cocaine as internal standard     

Flavonoids as chemotaxonomic markers for cultivated Amazonian coca

Purported ‘Amazonian coca,’ Erythroxylum coca var. ipadu Plowman (E. coca var. ipadu) was harvested from cultivated fields in Colombia, South America to determine: (a) its identity; (b) if its leaf flavonoids were complimentary to those present in leaf tissue of E. c. var. ipadu in our collection derived from Colombia; and (c) if complimentary, indicative of kinship to E. coca var. ipadu obtained from Colombia, or a related Erythroxylum taxon. Leaf extracts from Amazonian field-grown coca afforded eight O-conjugated flavonoids: two O-conjugates of taxifolin, one O-conjugate of quercetin, two O-conjugates of eriodictyol and three O-conjugates of kaempferol. Present also in leaf tissue of Amazonian field-grown coca, but lacking in leaf tissue from our collection of E. c. var. ipadu was an O-ethyl ester typically found in E. coca var. coca, kaempferols and a 7-O-rutinoside commonly encountered in the E. novogranatense taxons. Flavonoids found in our collection of E. coca var. ipadu were five O-conjugated derivatives of taxifolin and an O-conjugated quercetin. Leaf flavonoids of currently cultivated Amazonian coca are a mixture of those present in E. coca var. coca, E. coca var. ipadu and E. novogranatense var. truxillense, whereas those present in our authenticated living collection are derivatives of E. coca var. coca. Our data suggest that the Amazonian coca under cultivation in Colombia is a genetic hybrid cross between E. coca var. coca and E. novogranatense var. truxillense, occurring after 1972.     

Content and De Novo Synthesis of Cocaine in Embryos and Endosperms from Fruit of Erythroxylum coca Lam

There is controversy as to whether the mature fruit of Erythroxylumcoca var. coca Lam. contains the cocaine alkaloid (benzoylmethylecgonine).In the present study, cocaine was monitored to determine ifit was present in embryos and endosperms of mature fruit ofE. coca var. coca Lam., and if present, the time required forde novo synthesis in imbibing seed. Seeds from mature fruitof E. coca were dissected to separate the embryos from the endosperms.The separated embryos and endosperms were analysed for cocaine.Subsequently, endosperms and embryos from seed imbibed. undera light and dark treatment were separated on days 3, 6, 9, 12and 15 and analysed for cocaine. Cocaine was present in embryos(0.005% of d. wt) and endosperms (0–001% of d. wt) ofmature fruit of E. coca. De novo synthesis of cocaine occurredonly in embryos of seed imbibed under light after day 9 of imbibition. Erythroxylum coca, alkaloid, benzoylmethylecgonine, cocaine, embryo, endosperm, seed imbibition     

Variation of Alkaloid Content in Erythroxylum coca Leaves from Leaf Bud to Leaf Drop

A sequential study describing the content (%) of alkaloids inleaves of Erythroxylum coca var. coca Lam. from leaf bud developmentto leaf drop has not previously been conducted. The length oftime the leaf resides on the plant and its concurrent metabolicactivity also has not been defined. In the present study, cocaine,methyl ecgonine, hygrine, tropinone, trans -cinnamoylcocaine,cis-cinnamoylcocaine, tropacocaine and cuscohygrine were monitoredto determine: (1) their content and patterns of accumulationfrom leaf bud to leaf drop; (2) the time leaves resided on theplant; and (3) whether leaves were metabolically active untilleaf drop. E. coca plants were grown in a controlled environmentfor 37 months. Leaves similar in chronological age were extractedand analysed for alkaloid content by gas chromatography (GC)and gas chromatography/mass spectrometry (GC/MS). Cocaine washighest in 7 d old rolled leaves (0·75%) and declinedto 0·39% at leaf drop. Cocaine, methyl ecgonine, hygrine,tropinone, trans -cinnamoylcocaine, cis-cinnamoylcocaine, cuscohygrineand tropacocaine content in 35 d old (mature) leaves was 0·61,0·59, 0·68, 0·08, 0·31, 0·55,0·52, and 0·05%. respectively. Cocaine, methylecgonine, hygrine, cis -cinnamoylcocaine, and cuscohygrine displayeda gradual decline from week 2 to week 36 of leaf duration. Tropinoneand tropacocaine were the least abundant of the alkaloids monitored.Cis-cinnamoylcocaine content exceeded cocaine at week 12, 16,and weeks 19 to 23 of leaf duration. Trans -cinnamoylcocainewas highest in rolled leaves (week 1) and in expanded leavesafter week 30. The monitored alkaloids appeared to be part ofthe metabolically active pool of the leaf. Leaves remained onthe E. coca plants for 36 weeks.Copyright 1994, 1999 AcademicPress Cocaine, methyl ecgonine, hygrine, tropinone, trans-cinnamoylcocaine, cis-cinnamoylcocaine, cusco-hygrine, tropacocaine, leaf bud, rolled leaves, expanded leaves, alkaloids, patterns, fluctuation, Erythroxylum coca var. coca, E, coca     

Cocaine and Cinnamoylcocaine Content of Erythroxylum Species

The dried leaves of 31 species and two varieties of the genusErythroxylum were quantitatively analysed for cocaine and bothcis- and trans-cinnamoylocaine, using a stable-isotope dilutionmethod and gas chromatography-mass spectrometry selected-ionmonitoring. Both cocaine and cinnamoylcocaine were detectedin all four varieties of cultivated coca (E. coca var. coca,E. coca var. ipadu, E. novogranatense var. novogranatense andE. novogranatense var.truxillense). Cinnamoylcocaines (cis andtrans) were found in much higher concentrations in both varietiesof E. novogranatense than in either variety of E. coca. Amazoniancoca (E. coca var. ipadu) contained consistently lower cocainelevels than the montane variety (E. coca var. coca). Twenty-ninewild species of Erythroxylum, selected to represent morphological,ecological and taxonomic diversity, were analysed for the samealkaloids; cocaine was detected in 13 neotropical species, representingfive sections of the genus. No cocaine was detected in the OldWorld species. Two wild species from Venezuela, E. recurrensand E. steyermarkii, contained cocaine levels comparable tothose found in the commercially cultivated species. Erythroxylaceae, Erythroxylum, coca, cocaine, cinnamoylcocaine, alkaloids, chemotaxonomy     

The influence of bioturbation by the sandprawn Callianassa kraussi on feeding and survival of the bivalve Eumarcia paupercula and the gastropod Nassarius kraussianus

Field observations and experimental evidence have shown that bioturbation by the southern African sandprawn Callianassa kraussi may significantly influence the abundance and distribution of the filter-feeding bivalve Eumarcia paupercula and the grazing gastropod Nassarius kraussianus. It was hypothesized that (1) sediment reworking by C. kraussi negatively affects microalgal growth on the sediment surface, leading to a reduction in food intake by N. kraussianus, (2) sediment deposited by C. kraussi will also diminish the food uptake of E. paupercula by interfering with its filtration mechanism. To test these hypotheses, manipulative field and laboratory experiments were undertaken in which N. kraussianus and E. paupercula were added to treatments with and without C. kraussi, and their survival and gut chlorophyll-a content measured. The effects of C. kraussi on sediment erodability and on condition of E. paupercula (tissue mass/shell length) were determined in a second experiment. In the presence of C. kraussi, (1) microalgal consumption by both N. kraussianus and E. paupercula was halved; (2) condition and survival of E. paupercula were significantly reduced but survival of N. kraussianus was unaffected; and (3) sediment erodability was increased. A significant negative relationship was established between sediment erodability and condition of E. paupercula. Evidently C. kraussi exerts a strongly negative influence on the feeding of E. paupercula and N. kraussianus, and this may explain the scarcity of these organisms in areas containing high densities of C. kraussi.     

Palatability of the Antarctic rhodophyte Palmaria decipiens (Reinsch) RW Ricker and its endo/epiphyte Elachista antarctica Skottsberg to sympatric amphipods

Although many macroalgae that occur throughout Western Antarctic Peninsular waters are known to produce defensive secondary metabolites that deter grazing, the rhodophyte, Palmaria decipiens is palatable to several sympatric meso- and macro-grazers. It has been hypothesized that high levels of mesoherbivory by amphipods may account for the conspicuous lack of filamentous epiphytes emerging from the thalli of marcophytes in this region. Nonetheless, Elachista antarctica is a filamentous phaeophyte found growing within, and emerging from the thallus of the rhodophyte P. decipiens. It is surprising that E. antarctica occurs exclusively in association with a palatable species of macroalgae considering the standing biomass of other chemically defended unpalatable species is very high. We tested the hypothesis that E. antarctica grows on P. decipiens due to the host's overwhelming palatability compared to that of the epiphyte. That is, the hypothesis that mesograzers prefer the host over the epiphyte, grazing around emerging filaments. Choice and no choice feeding assays with live tissues of E. antarctica and P. decipiens were conducted in three different trials with four sympatric amphipod species (Prostebbingia gracilis, Gondogeneia antarctica, Oradarea bidentata, and Paraphimedia integricauda) commonly found in association with P. decipiens. G. antarctica consumed both species but ate P. decipiens at a faster rate than the epiphyte in two of three trials. P. gracilis, O. bidentata, and P. integricauda fed on the epiphyte, E. antarctica at faster rates than upon P. decipiens. Aggressive grazing of the epiphyte by this suite of amphipods indicates that differences in palatability and differences in grazing pressure on host and epiphyte do not explain the exclusive epiphytism of E. antarctica on P. decipiens.     

Cucurbitacin E and I in Iberis amara: Feeding inhibitors for Phyllotreta nemorum

Cucurbitacin E and cucurbitacin I have been isolated from green parts of Iberis amara and identified by TLC, UV and MS. It is shown that cucurbitacins act as feeding inhibitors for the flea beetle Phyllotreta nemorum. The most potent feeding inhibitors in green parts of I. amara towards P. nemorum are cucurbitacin E and I, and the concentrations of these compounds in the plant are found to be high enough to prevent feeding of the flea beetle.     

Botryosphaeriaceae from tuart (Eucalyptus gomphocephala) woodland, including descriptions of four new species

Eucalyptus gomphocephala (tuart) is a tree native to the southwest coast of Western Australia, where, in some areas, there is a significant decline in the health of tuart. Botryosphaeriaceous taxa have been isolated as endophytes and canker pathogens from numerous hosts in many parts of the world and have been implicated in the decline of E. gomphocephala. In the present study, endophytic fungi were isolated from a wide variety of native woody plant species (Acacia cochlearis, A. rostellifera, Allocasuarina fraseriana, Agonis flexuosa, Banksia grandis, E. gomphocephala, E. marginata and Santalum acuminatum), at two locations in native E. gomphocephala woodland; a site in decline at Yalgorup National Park and a healthy site at Woodman Point Regional Park. Of the 226 isolates obtained, 154 were botryosphaeriaceous taxa, 80 % of which were found to be Neofusicoccum australe, isolated from all hosts at both collection sites. Four new species are described, Dothiorella moneti, Dothiorella santali, Neofusicoccum pennatisporum, and a species belonging to a genus only recently included in the Botryosphaeriaceae, Aplosporella yalgorensis. The other species isolated were Botryosphaeria dothidea on the new hosts A. rostellifera, A. cochlearis and E. marginata and Dichomera eucalypti, on the new host E. marginata. None of the new species formed lesions on excised stems of their host species, E. gomphocephala, or a common plantation species, E. globulus. However, Neofusicoccum australe formed lesions on excised stems of E. globulus and E. gomphocephala.     

Infection of tomato by Tomato Yellow Leaf Curl Virus alters the foraging behavior and parasitism of the parasitoid Encarsia formosa on Bemisia tabaci

Encarsia formosa Gahan is a solitary endoparasitoid that is commercially reared and released for augmentative biological control of whiteflies including Bemisia tabaci (Gennadius). Bemisia tabaci biotypes B and Q are two most invasive species that greatly reduce crop yields in China by feeding on plant sap and by transmitting Tomato Yellow Leaf Curl Virus (TYLCV). The effects of TYLCV infection of tomato on E. formosa foraging on B. tabaci B and Q are unknown. In Y-tube olfactometer assays in the present study, E. formosa significantly preferred TYLCV-infected tomato plants over TYLCV-free plants. The wasp females also significantly preferred TYLCV-infected tomato plants infested with 3rd-instar nymphs of B. tabaci biotype Q over TYLCV-free plants with biotype Q nymphs. However, no significant differences were observed when B. tabaci biotype B was infested on tomato plants. The oviposition bioassays confirmed that TYLCV infection on tomato plants resulted in the recruitment of parasitoids. These results indicate that TYLCV-infection of tomato increase the foraging of E. formosa on B. tabaci, as differs on the B and Q biotypes.     

Phyllocarida and the origin of the Malacostraca

Tentative homologies with Decapoda are proposed for grooves on the carapace of Echinocaris and Montecaris. Together with F.R. Schram's hoploid trend, reinforced by the discovery of a raptorial limb in Sairocaris, this may strengthen a phyllocarid origin for Eumalacostraca. Both these trends, however, may be parallelisms, and phylogenetic analysis is required. Pleopods are here proved to have been present in additional Archaeostraca, which improves their ancestral condition. E. Dahl's view of the phyllocarids as an evolutionary dead end is falsified from fossils, and the phyllocarids are supported as a stem group of the Malacostraca.     

Cysteine proteinases substitute for serine proteinases in the midgut glands of Crangon crangon and Crangon allmani (Decapoda: Caridea)

The utilization of dietary proteins in crustaceans is facilitated by a set of peptide hydrolases which are often dominated by “trypsin-like” serine proteinases. As expected, the North Sea shrimps Crangon crangon and Crangon allmani showed in their midgut glands high proteolytic activities. However, the majority of animals lacked trypsin and chymotrypsin. Conversely, a minority of about 10% of the animals had elevated trypsin activities. The appearance of trypsin was neither related to the mode of feeding nor to the nutritive state of the animals. When present, trypsin was expressed in both species as a single isoform of apparently 20 kDa. The lack of serine proteinases was also confirmed by inhibitor assays. AEBSF, a serine proteinase inhibitor, slightly reduced total proteinase activity by less than 10%. In contrast E 64, a cysteine proteinase inhibitor, caused a reduction of more than 70% of total proteinase activity, indicating that a substantial share of proteolytic activity is caused by cysteine proteinases. Cathepsin L-like proteinases were identified as major cysteine proteinases.A comparison with the eucarid crustaceans Pandalus montagui, Pagurus bernhardus, Cancer pagurus and Euphausia superba showed a similar high level of total proteinase activity in all species. Trypsin, however, varied significantly between species showing lowest activities in Caridea and the highest activity in E. superba. E 64 suppressed total proteinase activity by more than 70% in Crangon species but not in C. pagurus and E. superba. In contrast, the serine proteinase inhibitor AEBSF had only little effect in Caridea but was most effective in P. bernhardus, C. pagurus and E. superba. The results may indicate different traits of food utilization strategies in some eucarid crustaceans. Caridea may express predominantly cysteine proteinase, while in Anomura, Brachyura and Euphausiacea, serine proteinases may prevail.     

Antifeedant activity of ethanolic extract from Flourensia oolepis and isolation of pinocembrin as its active principle compound

The ethanolic extract from Flourensia oolepis aerial parts showed strong antifeedant activity against the pest larvae, Epilachna paenulata, with an antifeedant index (AI%) of 99.1% at 100 μg/cm². Based on chromatographic fractionation of the extract, guided by bioassays on E. paenulata, the flavanone pinocembrin (1) was isolated as the most active principle. In a choice assay, compound 1 showed strong antifeedant activity against E. paenulata, Xanthogaleruca luteola and Spodoptera frugiperda with an AI% of 90, 94 and 91% (p < 0.01) respectively, at 50 μg/cm². The dosages necessary for 50% feeding inhibition of the insects (ED₅₀) were 7.98, 6.13 and 8.86 μg/cm², respectively. The feeding inhibitory activity of 1 against E. paenulata was compared with the activity of other structurally related flavonoids like naringenin, which was inactive up to 100 μg/cm², catechin which was nearly 6 times less active than 1, and quercetin which was equally active as 1. The effect of these on the feeding behavior of E. paenulata was also studied.     

Feeding and survival rates of the copepods Euterpina acutifrons Dana and Acartia grani Sars on the dinoflagellates Alexandrium minutum Balech and Gyrodinium corsicum Paulmier and the Chryptophyta Rhodomonas baltica Karsten

Here we studied the feeding performances and survival rates of the copepods Euterpina acutifrons and Acartia grani exposed to cultures of the toxic dinoflagellates Alexandrium minutum and Gyrodinium corsicum and the nontoxic Chryptophyta Rhodomonas baltica. The presence of hemolysins in G. corsicum was also analysed. For both copepod species, high ingestion rates on A. minutum and lower but similar rates on G. corsicum and R. baltica were found. After 120 h, only the E. acutifrons that grazed on G. corsicum or R. baltica showed 100% survival, whereas survival of the copepods that grazed on A. minutum was significantly lower. A. grani was more sensitive to the dinoflagellates, and at 120 h showed survival rates of 70% on G. corsicum and 60% on A. minutum. These results support the hypothesis that the feeding interactions between copepods and toxic dinoflagellates are highly species-specific. Long-term observations indicated that E. acutifrons can graze on G. corsicum without impairing their feeding or growth performances. Both copepod species consumed A. minutum in the short-term, but their long-term impact on this and other toxic marine dinoflagellates would be limited by the toxic effects of these flagellates. We did not find any evidence of haemolytic activity in extracts of G. corsicum. Toxins other than haemolysins may be responsible for the deleterious effects observed on A. grani.     

The steam volatile leaf oils of some species of Eucalyptus subseries Strictinae

The volatile leaf oils of Eucalyptus stenostoma, E. fraxinoides, E. triflora, E. dendromorpha, E. burgessiana, E. rupicola, E. approximans subsp. approximans, E. approximans subsp. codonocarpa and E. paliformis were qualitatively very similar to each other but significantly different from the leaf oils of the remaining species of the subseries. Their leaf oils were characterized by high concentrations of p-cymene and by the presence of α- and β-phellandrene, variable amounts of piperitone, trans- and cis-piperitol and trans- and cis-p-menth- 2-en-1-ol. The three last named alcohols have not been previously reported from the Myrtaceae.     

Content and Distribution of Erythroxylum coca Leaf Alkaloids

Fully expanded leaves of Erythroxylum coca var. coca Lam. (Erythroxylaceae)35-70-d-old were harvested from 21-month-old plants grown undergreenhouse conditions. Each harvested leaf was placed on a plexiglasssilhouette according to its dimensions and divided into fourprimary sections (petiole, base, mid and anterior) and threesubsections (lamina periphery, false mid-rib, and true mid-rib)to determine the distribution and content of hygrine, cuscohygrine,trans-cinnamoylcocaine, cis -cinnamoylcocaine, tropacocaine,tropinone, methyl ecgonine and cocaine. The leaf sub-sectionsand petiole were extracted and analysed for alkaloid content(%) by HPLC (cocaine alkaloid only) gas chromatography and GC/MS.Cocaine, methyl ecgonine and hygrine were highest in the laminaperiphery with a content of 0·48, 0·46 and 0·32%,respectively. Trans-cinnamoylcocaine was pre-eminent of thecinnamoylcocaines and was most abundant in the petiole at acontent of 0·24%. Cis-cinnamoylcocaine, tropinone, cuscohygrineand tropacocaine were ubiquitously distributed throughout theleaf where the average contents were 0·14, 0·004,0·16 and 0·04%, respectively.Copyright 1995, 1999Academic Press Alkaloids, Erythroxylum coca var. coca, E. coca, leaf alkaloids, petiole alkaloids, cocaine, cuscohygrine, methyl ecgonine, hygrine, tropinone, tropacocaine, trans-cinnamoylcocaine, cis-cinnamoylcocaine