Search Results (6)

Background: Primary amebic meningoencephalitis (PAM) caused by Naegleria fowleri is a rare and devastating infection of the central nervous system, often diagnosed late, due to its rapid progression and nonspecific symptoms. Case Presentation: We report one of the youngest documented pediatric Vietnamese cases of PAM in a 10-month-old girl from the Mekong Delta, Vietnam. The diagnosis was confirmed through multiplex real-time PCR (MPL-rPCR), microscopy, and sequencing. Clinical data were gathered retrospectively from medical records, and additional details were provided by the patient’s family. Treatment regimens, disease progression, and diagnostic challenges were reviewed and compared to existing literature. With intensive treatment, the child survived for 14 days, representing one of the longest reported pediatric PAM survival durations. No direct exposure to untreated freshwater or other typical risk factors for Naegleria fowleri infection was identified, underscoring the unique epidemiological nature of this case. MPL-rPCR enabled timely detection of the pathogen and demonstrated its utility in resource-limited settings. Conclusions: This case highlights the critical need for rapid, accessible diagnostic tools such as MPL-rPCR, particularly in resource-constrained environments where traditional diagnostics may not be feasible. It also emphasizes the importance of international collaboration and investment in cost-effective diagnostics and novel therapeutic strategies. The geographical expansion of PAM due to climate change further underscores the urgency of these measures to improve health outcomes in vulnerable populations. Full article

Naegleria fowleri is a pathogenic free-living amoeba, commonly found around the world in warm, fresh water and soil. N. fowleri trophozoites can infect humans by entering the brain through the nose and causing usually fatal primary amebic meningoencephalitis (PAM). Trophozoites can encyst to survive under unfavorable conditions such as cold temperature, starvation, and desiccation. Recent technological advances in genomics and bioinformatics have provided unique opportunities for the identification and pre-validation of pathogen-related and environmental resistance through improved understanding of the biology of pathogenic N. fowleri trophozoites and cysts at a molecular level. However, genomic and transcriptomic data on differential expression genes (DEGs) between trophozoites and cysts of N. fowleri are very limited. Here, we report transcriptome Illumina RNA sequencing (RNA-seq) for N. fowleri trophozoites and cysts and de novo transcriptome assembly. RNA-seq libraries were generated from RNA extracted from N. fowleri sampled from cysts, and a reference transcriptome was generated through the assembly of trophozoite data. In the database, the assembly procedure resulted in 42,220 contigs with a mean length of 11,254 nucleotides and a C+G content of 37.21%. RNA sequencing showed that 146 genes in cysts of N. fowleri indicated 2-fold upregulation in comparison with trophozoites of N. fowleri, and 163 genes were downregulated; these genes were found to participate in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The KEGG pathway included metabolic (131 sequences) and genetic information processing (66 sequences), cellular processing (43 sequences), environmental information processing (22 sequences), and organismal system (20 sequences) pathways. On the other hand, an analysis of 11,254 sequences via the Gene Ontology database showed that their annotations contained 1069 biological processes including the cellular process (228 sequences) and metabolic process (214 sequences); 923 cellular components including cells (240 sequences) and cell parts (225 sequences); and 415 molecular functions including catalytic activities (195 sequences) and binding processes (186 sequences). Differential expression levels increased in cysts of N. fowleri compared to trophozoites of N. fowleri, which were mainly categorized as serine/threonine protease, kinase, and lipid metabolism-related proteins. These results may provide new insights into pathogen-related genes or environment-resistant genes in the pathogenesis of N. fowleri. Full article

Swimming pool water treatment by chemicals is an essential step to avoid microbial proliferation and infections namely caused by free living amoeba such as, for example, primary amebic meningoencephalitis and Acanthamoeba keratitis. In the present study, a commercial reactive, CLORICAN, based on chlorine dioxide, was evaluated against Acanthamoeba spp. and Naegleria fowleri. We observed that CLORICAN could eliminate in a short period of incubation time both amoebae. Furthermore, Naegleria fowleri’s trophozoites were more sensitive than those of Acanthamoeba spp. By means of inverted microscopy, the chlorine dioxide was found to greatly affect morphology shape by increasing the cell size shrinkage. Full article

Free-living amoeba Naegleria fowleri causes a rapidly fatal infection primary amebic meningoencephalitis (PAM) in children. The drug of choice in treating PAM is amphotericin B, but very few patients treated with amphotericin B have survived PAM. Therefore, development of efficient drugs is a critical unmet need. We identified that the FDA-approved pitavastatin, an inhibitor of HMG Co-A reductase involved in the mevalonate pathway, was equipotent to amphotericin B against N. fowleri trophozoites. The genome of N. fowleri contains a gene encoding protein farnesyltransferase (FT), the last common enzyme for products derived from the mevalonate pathway. Here, we show that a clinically advanced FT inhibitor lonafarnib is active against different strains of N. fowleri with EC₅₀ ranging from 1.5 to 9.2 µM. A combination of lonafarnib and pitavastatin at different ratios led to 95% growth inhibition of trophozoites and the combination achieved a dose reduction of about 2- to 28-fold for lonafarnib and 5- to 30-fold for pitavastatin. No trophozoite with normal morphology was found when trophozoites were treated for 48 h with a combination of 1.7 µM each of lonafarnib and pitavastatin. Combination of lonafarnib and pitavastatin may contribute to the development of a new drug regimen for the treatment of PAM. Full article

During the first trimester of 2020, the Ministry of Health of Costa Rica reported the first three cases of primary amebic meningoencephalitis (PAM). In two cases, laboratory personnel of the hospitals preliminarily identified amoeboid forms in cerebrospinal fluid (CSF) samples. For the molecular confirmation of species, CSF samples were sent to our laboratory. We carried out microscopic analyses and exflagellation assays. Besides, samples were cultured in 2% casein hydrolysate medium and in non-nutrient agar plates supplemented with Escherichia coli. Finally, PCR and sequencing were employed for the molecular diagnosis and species identification. In all cases, the presence of Naegleria fowleri was confirmed. An environmental investigation to identify the possible infection sources was also performed. Water samples from hot springs and groundwater from an artisan well were collected and after filtration and culture in non-nutrient agar plates supplemented with E. coli, thermotolerance and exflagellation assays were carried out. For the positive samples, PCR and sequencing were performed, confirming the presence of N. fowleri in several water samples. The report of these cases and the possible association with hot springs has had a significant impact on the population and health authorities of Costa Rica. Full article

Free-living amoebae (FLA) can be found both in natural aquatic environments and in artificial, man-made aquatic environments. For a long time, (naked) FLA were considered to be harmless protozoa of soil and water. However, research since the 1960s has demonstrated that FLA can be pathogenic to humans and animals with nearly 100% morbidity from some strains. As etiological agents of the so-called Acanthamoebiasis, the Acanthamoebae can trigger several specific diseases (or symptoms) in humans. The amoebiasis of the central nervous system is called granulomatous amebic encephalitis (GAE), when Acanthamoebae are the etiological agents, showing subacute to chronic progress. GAE differs clinically from the primary amoebic meningoencephalitis (PAM(E)), which is caused by Naegleria fowleri. The Acanthamoeba keratitis is not necessarily associated with an immune suppression, but rather with a trauma, exposure to contaminated water or, particularly, the improper handling of contact lenses, which promotes infection. The clinical picture of GAE by Balamuthia mandrillaris is characterized by headache and neck stiffness. The infection is chronic and the time between infection and appearance of neurological symptoms may range from one month to about two years. The prevalence of FLA in water networks is associated with biofilms, where the amoebae live within a biocoenosis sympatric with other microorganisms. These biofilms serve as feeding grounds for the FLA and provide protection to a certain degree while the FLA adhere to the surfaces. In such a biocoenosis there are multiple interactions between FLA and other microorganisms: In addition to their role as pathogens, FLA are known to serve as host of and vehicles for diverse intracellular organisms (bacteria, viruses, eucaryonts), some of them being natural human pathogens. They act as reservoir or vehicle for various microorganisms such as various Legionella sp., Listeria monocytogenes, Pseudomonas aeruginosa and some Mycobacterium sp. In the cyst-stage of the FLA these intracellular organisms (endocytobionts) are protected to a high degree against any adverse environment (FLA as “Trojan horse”). This host-endocytobiont relationship may further lead to health risks in terms of the development of pathogenicity/virulence and antibiotic resistance (FLA as “Trainings ground”). Full article