Impact of inter-amoebic phagocytosis on the L. pneumophila growth.

Free-living amoebae are known to act as replication niches for the pathogenic bacterium Legionella pneumophila in freshwater environments. However, we previously reported that some strains of the Willaertia magna species are more resistant to L. pneumophila infection and differ in their ability to support its growth.

Endocannabinoids inhibit the growth of free-living amoebae.

The cannabinoid Delta(9)-tetrahydrocannabinol inhibits the growth of some pathogenic amoebae in vitro and exacerbates amoebic encephalitis in animal models. However, the effects of endogenous cannabinoids on amoebae remain unknown. Therefore, we tested several endocannabinoids (N-acyl ethanolamines and 2-O-acyl glycerol) on different genera of amoebae.

[Molecular diagnosis of a fatal primary amoebic meningoencephalitis in Guadeloupe (French West Indies)].

We report the first case of primary amoebic meningoencephalitis in a 9-year-old boy in Guadeloupe. The outcome was rapidly fatal in 7 days. The patient presumably acquired the infection by swimming and diving in a basin supplied by natural thermal water 1 week before onset of the disease.

Free-living freshwater amoebae differ in their susceptibility to the pathogenic bacterium Legionella pneumophila.

Legionella pneumophila is known as a facultative intracellular parasite of free-living soil and freshwater amoebae, of which several species have been shown to support the growth of the pathogenic bacteria. We report for the first time the behaviour of two strains (c2c and Z503) of the amoeba Willaertia magna towards different strains of L. pneumophila serogroup 1 and compared it with Acanthamoeba castellanii and Hartmannella vermiformis, known to be L.

An enzyme-linked immunosorbent assay (ELISA) for the identification of Naegleria fowleri in environmental water samples.

Naegleria fowleri, a free-living amoeba, is the causative agent of primary amoebic meningoencephalitis, a fatal human disease of the central nervous system often contracted after swimming in fresh water. Identifying sites contaminated by N. fowleri is important in order to prevent the disease.

Elimination of free-living amoebae in fresh water with pulsed electric fields.

This study investigates the effects of pulsed electric fields on the inactivation of trophozoite form of Naegleria lovaniensis Ar9M-1 in batch and flow processes, systematically examining the lethal effect of field strength, pulse duration, number of pulses, and pulse frequency. Our results show that amoebae eradication is modulated by pulse parameters, composition of the pulsing medium, and physiological state of the cells. Cell survival is not related to the energy delivered to the cell suspension during the electrical treatment.

Rapid detection and enumeration of Naegleria fowleri in surface waters by solid-phase cytometry.

A new method for the rapid and accurate detection of pathogenic Naegleria fowleri amoebae in surface environmental water was developed. The method is based on an immunofluorescent assay combined with detection by solid-phase cytometry. In this study we developed and compared two protocols using different reporter systems conjugated to antibodies.

Use of multiplex PCR and PCR restriction enzyme analysis for detection and exploration of the variability in the free-living amoeba Naegleria in the environment.

A multiplex PCR was developed to simultaneously detect Naegleria fowleri and other Naegleria species in the environment. Multiplex PCR was also capable of identifying N. fowleri isolates with internal transcribed spacers of different sizes.

The amoeba-to-flagellate transformation test is not reliable for the diagnosis of the genus Naegleria. Description of three new Naegleria spp.

Trophozoites of several isolates from one location in Australia have failed consistently to transform into flagellates, although they display all other characteristics of the genus Naegleria. When changing the standard transformation test, flagellates were produced. In phylogenetic trees derived from partial small subunit ribosomal DNA (SSUrDNA) sequences, one of these strains branches close to a cluster comprising N.

Assessing the risk of primary amoebic meningoencephalitis from swimming in the presence of environmental Naegleria fowleri.

Free-living Naegleria fowleri amoebae cause primary amoebic meningoencephalitis (PAM). Because of the apparent conflict between their ubiquity and the rarity of cases observed, we sought to develop a model characterizing the risk of PAM after swimming as a function of the concentration of N. fowleri.

Different growth rates in amoeba of genotypically related environmental and clinical Legionella pneumophila strains isolated from a thermal spa.

Two cases of legionellosis occurring 3 years apart were acquired in the same French thermal spa and were apparently due to the same strain of Legionella pneumophila serogroup 1, as shown by genomic macrorestriction analysis. Minor differences between the two isolates were found by random amplification PCR profiling which showed an additional band with one of the isolates. Analysis of 107 L.

Species specificity of a monoclonal antibody produced to Naegleria fowleri and partial characterization of its antigenic determinant.

Monoclonal antibody (Mab) 5D12 against Naegleria fowleri was analyzed for species specificity. Mab 5D12 reacted with a ubiquitous epitope present on the membrane of N. fowleri but not with soluble antigens.

Analysis of the 5.8S rRNA gene and the internal transcribed spacers in Naegleria spp. and in N. fowleri.

Internal transcribed spacers (ITS) and the 5.8S ribosomal gene of 21 Naegleria fowleri strains and eight other species including Naegleria gruberi were sequenced. The results showed that this region can help differentiate between and within species.

Genetic variation in the free-living amoeba Naegleria fowleri.

In this study, 30 strains of the pathogenic free-living amoeba Naegleria fowleri were investigated by using the randomly amplified polymorphic DNA (RAPD) method. The present study confirmed our previous finding that RAPD variation is not correlated with geographical origin. In particular, Mexican strains belong to the variant previously detected in Asia, Europe, and the United States.

Comparative recoveries of Naegleria fowleri amoebae from seeded river water by filtration and centrifugation.

Detection of pathogenic Naegleria fowleri in environmental water samples, which is necessary for the prevention of primary amoebic meningoencephalitis, generally requires concentrating the samples. Two concentration techniques, filtration and centrifugation, were used to study the recovery of N. fowleri, in vegetative or cystic form, that had been mixed with the two other thermotolerant Naegleria species, N.

Use of monoclonal antibodies to distinguish pathogenic Naegleria fowleri (cysts, trophozoites, or flagellate forms) from other Naegleria species.

Monoclonal antibodies (MAbs) reactive to the pathogenic amoeba Naegleria fowleri were analyzed by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay, Western blotting (immunoblotting), and radioimmunoprecipitation assay (RIPA). Two MAbs (3A4 and 5D12) showed reactivity by ELISA with all N. fowleri strains tested and no reactivity with the five other Naegleria species, N.

[Therapeutic evaluation of Mormoiron attapulgite in acute diarrheas of infants and children. A multicenter study in controlled liberal practice versus placebo in 113 patients].

A placebo-controlled multicenter study was carried out to evaluate the effectiveness and safety of Actapulgite in infants and children with acute diarrhea. One hundred and thirteen patients, with a mean age of 28 months, were enrolled by pediatricians in the office. As recommended during acute diarrhea, the dosage of Actapulgite was doubled during the first three treatment days (2 to 4 packets a day according to the patient's weight), and reduced by half thereafter.

Genetic structure of natural populations of the free-living amoeba, Naegleria lovaniensis. Evidence for sexual reproduction.

The genetic structure of two populations of Naegleria lovaniensis, comprising 71 isolates collected from the same local geographical area was investigated by isoenzyme analysis. Allelic variation at seven polymorphic enzymatic loci allowed identification of 45 distinctive genotype associations. Analysis of single locus variation reveals that most of them are close to Hardy-Weinberg equilibrium, which indicates segregation and free recombination between alleles.

[Acanthamoeba keratitis. Morphologic and isoenzymatic study of a strain isolated from a medically treated case].

We report here the third case of Acanthamoeba keratitis discovered in France in a 30 year-old man wearing soft contact lenses. Amoebas of Acanthamoeba genus could be isolated twice from his corneal ulcer with simple ocular swabs. Owing to medical treatment alone, a successful healing of his eye was obtained.

Appearance in Europe of Naegleria fowleri displaying the Australian type of restriction-fragment-length polymorphism.

We report for the first time the isolation in Europe of Naegleria fowleri showing a type of restriction-fragment-length polymorphism (RFLP) usually found in Australia. The presence of this type as well as the European type fluctuated with time in the cooling waters of the nuclear power station investigated. Two possible explanations for the appearance of the Australian N.

Large genetic heterogeneity within amoebas of the species Naegleria gruberi and evolutionary affinities to the other species of the genus.

The allozyme survey was extended to 7 strains of Naegleria gruberi and N. jadini in order to further characterize the genetic structure of these free-living amoebas. As formerly known for several characters the electrophoretic evidence reveals considerable heterogeneity at the genetic level among N.

Application of isoenzymatic typing to the identification of nonaxenic strains of Naegleria (Protozoa, Rhizopoda).

Isoenzymatic typing of the different species of Naegleria was studied by comparing isoelectric focusing on axenic and nonaxenic strains using the most discriminating enzymes: lactate dehydrogenase (LDH), malic enzyme (ME), beta-hydroxybutyrate dehydrogenase (beta-HBDH), superoxide dismutase (SOD), and acid phosphatase (AP). The results show that bacteria in nonaxenic cultures have no influence on the interpretation of zymograms. The specificity of the enzymatic patterns of each strain is maintained in all of these enzymatic systems except LDH.

First evidence for diploidy and genetic recombination in free-living amoebae of the genus naegleria on the basis of electrophoretic variation.

Electrophoretic variation for 15 enzyme-coding genes was studied in various Naegleria (Rhizopoda, Vahlkampfiidae) species. The occurrence of complex banding patterns provided the first evidence of a diploid structure of the genome of these amoebae. The putative loci identified were found not to be linked and the genotypic distribution suggested chromosomal recombination for one species (Naegleria lovaniensis).