Jean-Marie Bastide

Composition and antimalarial activity in vitro of volatile components of lippia multiflora

The essential oil of Lippia multiflora was prepared by hydrodistillation of leaves and stalks and characterized by GC and mass spectroscopy. The oil was tested for antimalarial activity on in vitro cultures of Plasmodium falciparum (FcB1-Columbia chloroquine-resistant strain and F32-Tanzania chloroquine-sensitive strain). The dilutions inhibiting the in vitro growth of the parasite by 50% 24 and 72 hr after administration of the essential oil to the parasite culture were 1/12,000 and 1/21,000, respectively. When tested on a highly synchronized culture, the essential oil inhibited growth mostly at the trophozoite-schizont step, indicating a potential effect on the first nuclear division of the parasite.

In vitro antimalarial activity of eight essential oils

Abstract Essential oils of eight plants were prepared by hydrodistillation of leaves and twigs (stems) of Artemisia vulgaris, Eucalyptus globulus, Myrtus communis, Juniperus communis, Lavandula angusti/olia, Origanum vulgare, Rosmarinus officinalis and Salvia officinalis. Their oil components were characterized by GC and MS. These oils were then tested for in vitro antimalarial activity on Plasmodium falciparum. Two strains of P. falciparum were tested: FcBl-Columbia (chloroquine-resistant) and a Nigerian chloroquine-sensitive strain. Concentrations inhibiting 50% of the parasite in vitro growth, obtained after 24 and 72 h contact between the oil and the parasite culture, ranged from 150 μg/mL to 1 mg/mL. The best results were obtained with Myrtus communis and Rosmarinus officinalis oils which inhibited P. falciparum at concentrations ranging from 150 to 270 μg/mL.

In vitro antiplasmodial activity of stem and root extracts of Nauclea latifolia S.M. (Rubiaceae).

Aqueous extracts from Nauclea latifolia S.M. (Rubiaceae), a plant commonly used in Ivory Coast by traditional healers for the treatment of malaria, were tested on two strains of Plasmodium faliparum: FcB1-Colombia (chloroquine-resistant) and a Nigerian strain (chloroquine-sensitive). The extracts were obtained from stems and roots of the plant in two forms, infusion and decoction, both methods used by most traditional healers. The in vitro activity of N. latifolia extracts on P. falciparum was assessed both visually and by a radioactive method. The visual analysis allowed determination of the time of extract action on the erythrocytic cycle, as well as the parasitic stage of most inhibitory effect. Similar results were obtained applying fresh, frozen or lyophilized extracts. The IC50 values determined were within the range already reported for other antimalarial plants such as Azadirachta indica A. Juss (Meliaceae) or Artemisia annua L. (Asteraceae). Aqueous extracts of N. latifolia inhibited P. falciparum (FcB1 strain) mainly at the end of the erythrocytic cycle (32nd to 48th hour).

Genetic Polymorphism of Aspergillus fumigatus in Clinical Samples from Patients with Invasive Aspergillosis: Investigation Using Multiple Typing Methods

ABSTRACT The genotypes of 52 strains of Aspergillus fumigatusisolated from 12 patients with invasive aspergillosis were investigated using three typing methods (random amplified polymorphic DNA, sequence-specific DNA polymorphism, and microsatellite polymorphism) combined with multilocus enzyme electrophoresis. Isolates were from patients hospitalized in three different geographic areas (Lyon, France; Grenoble, France; and Milan, Italy). In each case, the genetic polymorphism of several colonies (two to five) within the first respiratory clinical sample was studied. For the 52 isolates tested, random amplified polymorphic DNA identified 8 different genotypes, sequence-specific DNA polymorphism identified 9 different types, and microsatellite polymorphism identified 14 types. A combination of these results with multilocus enzyme electrophoresis study identified 25 different types within the sample studied. We identified 3 patients (of the 12 studied) who carried a single genotype; 6 patients were infected by two genotypes, 1 patient had four genotypes, while the last patient had five. A combination of typing methods provided better discrimination than the use of a single method. Typing methods revealed a population structure within each geographical site, suggesting that the epidemiology of A. fumigatus should be considered separately for each of these geographic areas. This study demonstrates the usefulness of combining several typing methods in reaching an understanding of the epidemiology of A. fumigatus and clarifies whether it is sufficient to type one isolate from each specimen to determine the strain involved in invasive aspergillosis.

Atypical strains of Candida albicans recovered from AIDS patients

By using multilocus enzyme electrophoresis (MLEE) we have analyzed the genetic diversity encountered among chlamydospore-positive Candida albicans strains. While the type II strains of the former C. stellatoidea were genetically indistinguishable from those of C. albicans, type I strains constituted a distinct subgroup compared with C. albicans strains. Nevertheless, all these strains remained genetically very closely related compared with other species of Candida (e.g. C. tropicalis, C. krusei and C. glabrata). These results corroborate the synonymy between C. stellatoidea and C. albicans. Chlamydospore-positive C. albicans strains with atypical sugar assimilation patterns displayed a great genetic divergence from the cluster constituted by C. albicans and the strains of the former C. stellatoidea. However, these atypical strains were more closely related to C. albicans than they were to C. tropicalis, C. krusei or C. glabrata. These strains represent a genetically entity distinct from the typical C. albicans strains used in this study. The data also support the view that the atypical strains described here belong to the same genetic group as atypical C. albicans strains previously described by others.

Typing of Saccharomyces cerevisiae Clinical Strains by Using Microsatellite Sequence Polymorphism

ABSTRACT It seems that S. cerevisiae, which was thought for about 30 years to be a nonpathogenic yeast, should now be considered an opportunistic pathogen. In this study, we estimated the discrimination ability of the microsatellite sequence amplification technique within a sample of clinical and reference S. cerevisiae strains and S. boulardii reference strains.

Genetic Structure of Candida glabrata Populations in AIDS and Non-AIDS Patients

ABSTRACT The genotypes of 63 strains (11 reference strains and 52 strains from hospitalized patients) of the haploid yeast Candida glabrata were determined from 33 putative gene enzymatic loci. This enabled the characterization of 26 different multilocus genotypes. Genetic differentiation was found between distant hospitals (located in Montpellier and Paris, France) but not for other parameters (anatomic origins or human immunodeficiency virus-positive [HIV+] and HIV− patients). Strong nonrandom association between loci could be seen. Such statistical linkages were confirmed upon comparing the patterns of 14 RAPD [random(ly) amplified polymorphic DNA] primers from 20 of these strains to results obtained from multilocus enzyme electrophoresis analysis. This finding suggests a mainly clonal mode of reproduction of C. glabrata. The consequences of the clonality displayed by C. glabrata populations on the epidemiology of this yeast are also discussed.

Combination of three typing methods for the molecular epidemiology of Aspergillus fumigatus infections

This study investigated the source of infection and strain relatedness of Aspergillus fumigatus isolates from bronchial colonisation and invasive aspergillosis (IA) in four transplant patients. Environmental isolates from the patients home and from the hospital and infecting isolates were obtained for patient A who developed IA. Clinic environmental and colonising isolates were obtained for patient B. Sequential isolates were obtained from various organs from patient C who developed IA and also from patient D who had a bronchitic aspergillosis that developed into IA. Ninety-one A. fumigatus isolates were analysed by three typing methods: multi-locus enzyme electrophoresis (MLEE), random amplified polymorphic DNA (RAPD) and sequence-specific DNA primers (SSDP). The three combined typing methods demonstrated a greater differentiation of isolates than the typing methods used separately or in pairs. This demonstrated the genotypic variability of A. fumigatus and facilitated better epidemiological analysis. Large polymorphisms were demonstrated for each patient isolate between and colonies within various samples. The relatedness of the isolates suggested nosocomially acquired aspergillosis for patient B, but the source of infection for patient A remained unclear. The results suggested at least three multiple infections among the four patients. This study enabled the identification of the source of infection and strain relatedness, which in turn facilitates the development of preventive measures for patient management in the future.

Genetic diversity in the yeast species Malassezia pachydermatis analysed by multilocus enzyme electrophoresis

Fifty-two strains of the yeast species Malassezia pachydermatis were analysed by multilocus enzyme electrophoresis. M. pachydermatis appeared to be genetically heterogeneous. A total of 27 electrophoretic types were identified that could be divided into five distinct groups with different host specificities. The diversity revealed by this electrophoretic method matched remarkably well the reported genetic variability obtained by comparing large subunit rRNA sequences. This study also suggests that genetic exchanges can occur in the anamorphic species M. pachydermatis.

Multilocus enzyme electrophoresis analysis of Aspergillus fumigatus strains isolated from the first clinical sample from patients with invasive aspergillosis

The genotypes of 50 isolates of Aspergillus fumigatus from 11 patients with invasive aspergillosis, obtained from three hospitals in different geographical areas, were determined by multilocus enzyme electrophoresis (MLEE). The study analysed the genetic polymorphism of multiple isolates from the first sample. Seven of the 14 enzymic loci studied were polymorphic, giving rise to eight different electrophoretic types. For nine of 11 patients studied, no polymorphism was observed in isolates within the first clinical sample. Analysis of genetic distance between electrophoretic types demonstrated a genetic heterogeneity within each geographical site. Moreover, some genotypes were preferentially found in a given area and this revealed a population structure within these geographical sites. Therefore, the epidemiology of A. fumigatus should be considered separately for each of these areas. The multiple discriminatory markers of MLEE seem to provide a powerful tool for increasing the understanding of the biology of this fungus.