Fossil Geyserite and Testate Amoebae in Geothermal Spring Vent Pools: Paleoecology and Variable Preservation Quality in Jurassic Sinter of Patagonia (Deseado Massif, Argentina).

Geyserite is a type of terrestrial siliceous hot spring deposit (sinter) formed subaerially in proximal vent areas, with near-neutral pH, alkali chloride discharge fluids characterized by initial high temperatures (~73°C to up to 100°C) that fluctuate rapidly in relation to dynamic hydrology, seasonality, wind, and other environmental parameters. We analyzed sinters at the Claudia paleogeothermal field from the Late Jurassic (~150 Ma) Deseado Massif geological province, Argentinean Patagonia. The geyserite samples-with spicular to columnar to nodular morphologies-contain abundant microfossils in monotypic assemblages that occur in three diagenetic states of preservation. The best-preserved microfossils consist of vesicle-like structures with radial heteropolar symmetry (~35 µm average diameter), circular apertures, smooth walls lacking ornamentation, and disk- or beret-like shapes. Comparisons with extant, morphologically similar organisms suggest an affinity with the testate amoebae of the Arcella hemisphaerica-Arcella rotundata complex and Centropyxis aculeata strain discoides. These species occur in active geothermal pools between 22°C and 45°C, inconsistent with the temperature of formation of modern geyserites. We propose that the testate amoebae may have colonized the geyserite during cooler phases in between spring-vent eruptive cycles to prey on biofilms. Silica precipitation through intermittent bathing and splashing of fluctuating thermal fluid discharge could have led to their entrapment and fossilization. Petrographic analysis supports cyclicity in paleovent water eruptions and later diagenesis that transformed the opal into quartz. Spatially patchy degradation and modification of the silicified microorganisms resulted in variable preservation quality of the microfossils. This contribution illustrates the importance of microscale analysis to locate early silicification and identify high-quality preservation of fossil remains in siliceous hot spring deposits, which are important in early life studies on Earth and potentially Mars.

Neutrophils versus Protozoan Parasites: Plasmodium, Trichomonas, Leishmania, Trypanosoma, and Entameoba.

Neutrophils are the most abundant polymorphonuclear granular leukocytes in human blood and are an essential part of the innate immune system. Neutrophils are efficient cells that eliminate pathogenic bacteria and fungi, but their role in dealing with protozoan parasitic infections remains controversial. At sites of protozoan parasite infections, a large number of infiltrating neutrophils is observed, suggesting that neutrophils are important cells for controlling the infection. Yet, in most cases, there is also a strong inflammatory response that can provoke tissue damage. Diseases like malaria, trichomoniasis, leishmaniasis, Chagas disease, and amoebiasis affect millions of people globally. In this review, we summarize these protozoan diseases and describe the novel view on how neutrophils are involved in protection from these parasites. Also, we present recent evidence that neutrophils play a double role in these infections participating both in control of the parasite and in the pathogenesis of the disease.

Unusual and Unconsidered Mechanisms of Bacterial Resilience and Resistance to Quinolones.

Quinolone resistance has been largely related to the presence of specific point mutations in chromosomal targets, with an accessory role of impaired uptake and enhanced pump-out. Meanwhile the relevance of transferable mechanisms of resistance able to protect the target of pump-out or inactivate quinolones has been increasingly reported since 1998. Nevertheless, bacteria have other strategies and mechanisms allowing them to survive and even proliferate in the presence of quinolones, which might be qualified as resistance or resilience mechanisms. These include decreasing levels of quinolone target production, transient amoeba protection, benthonic lifestyle, nutrient-independent slow growth, activation of stringent response, inactivation or degradation of quinolones as well as apparently unrelated or forgotten chromosomal mutations. These mechanisms have been largely overlooked, either because of the use of classical approaches to antibiotic resistance determination or due to the low increase in final minimum inhibitory concentration levels. This article is devoted to a review of a series of these mechanisms.

Attenuation of In Vitro and In Vivo Virulence Is Associated with Repression of Gene Expression of AIG1 Gene in Entamoeba histolytica.

Entamoeba histolytica virulence results from complex host-parasite interactions implicating multiple amoebic components (e.g., Gal/GalNAc lectin, cysteine proteinases, and amoebapores) and host factors (microbiota and immune response). UG10 is a strain derived from E. histolytica virulent HM-1:IMSS strain that has lost its virulence in vitro and in vivo as determined by a decrease of hemolytic, cytopathic, and cytotoxic activities, increased susceptibility to human complement, and its inability to form liver abscesses in hamsters. We compared the transcriptome of nonvirulent UG10 and its parental HM-1:IMSS strain. No differences in gene expression of the classical virulence factors were observed. Genes downregulated in the UG10 trophozoites encode for proteins that belong to small GTPases, such as Rab and AIG1. Several protein-coding genes, including iron-sulfur flavoproteins and heat shock protein 70, were also upregulated in UG10. Overexpression of the EhAIG1 gene (EHI_180390) in nonvirulent UG10 trophozoites resulted in augmented virulence in vitro and in vivo. Cocultivation of HM-1:IMSS with E. coli O55 bacteria cells reduced virulence in vitro, and the EhAIG1 gene expression was downregulated. In contrast, virulence was increased in the monoxenic strain UG10, and the EhAIG1 gene expression was upregulated. Therefore, the EhAIG1 gene (EHI_180390) represents a novel virulence determinant in E. histolytica.

Diversity of Surface Fibril Patterns in Mimivirus Isolates.

Among the most intriguing structural features in the known virosphere are mimivirus surface fibrils, proteinaceous filaments approximately 150 nm long, covering the mimivirus capsid surface. Fibrils are important to promote particle adhesion to host cells, triggering phagocytosis and cell infection. However, although mimiviruses are one of the most abundant viral entities in a plethora of biomes worldwide, there has been no comparative analysis on fibril organization and abundance among distinct mimivirus isolates. Here, we describe the isolation and characterization of Megavirus caiporensis, a novel lineage C mimivirus with surface fibrils organized as "clumps." This intriguing feature led us to expand our analyses to other mimivirus isolates. By employing a combined approach including electron microscopy, image processing, genomic sequencing, and viral prospection, we obtained evidence of at least three main patterns of surface fibrils that can be found in mimiviruses: (i) isolates containing particles with abundant fibrils, distributed homogeneously on the capsid surface; (ii) isolates with particles almost fibrilless; and (iii) isolates with particles containing fibrils in abundance, but organized as clumps, as observed in Megavirus caiporensis. A total of 15 mimivirus isolates were analyzed by microscopy, and their DNA polymerase subunit B genes were sequenced for phylogenetic analysis. We observed a unique match between evolutionarily-related viruses and their fibril profiles. Biological assays suggested that patterns of fibrils can influence viral entry in host cells. Our data contribute to the knowledge of mimivirus fibril organization and abundance, as well as raising questions on the evolution of those intriguing structures. IMPORTANCE Mimivirus fibrils are intriguing structures that have drawn attention since their discovery. Although still under investigation, the function of fibrils may be related to host cell adhesion. In this work, we isolated and characterized a new mimivirus, called Megavirus caiporensis, and we showed that mimivirus isolates can exhibit at least three different patterns related to fibril organization and abundance. In our study, evolutionarily-related viruses presented similar fibril profiles, and such fibrils may affect how those viruses trigger phagocytosis in amoebas. These data shed light on aspects of mimivirus particle morphology, virus-host interactions, and their evolution.

Identificación de amebas de vida libre en tanques de almacenamiento de agua de edificios del área metropolitana de Ambato / Identification of free-living amoebas in water storage tanks of buildings in the metropolitan area of Ambato

Naegleria fowleri, Acanthamoeba spp. y Balamuthia mandrillaris, son especies de ameba de vida libre (AVL) que causan enfermedades en el sistema nervioso central grave y puede ser fatal en humanos y otros animales. Todos los géneros de AVL tienen dos etapas: quiste y trofozoíto. Los trofozoítos se dividen por fisión binaria, se alimentan activamente y es la etapa infecciosa del organismo. En el medio ambiente, los acanthopodios permiten que los trofozoítos se adhieran a las superficies de las bacterias, hongos, algas y detritos en los que se encuentran su alimento. Acanthamoeba spp. Parecen mantener una relación endosimbiótica con algunas de las bacterias que consumen. Muchas de estas bacterias, como la Legionella, Pseudomonas, Vibrio y algunas micobacterias parecen adquirir factores de virulencia mientras están viviendo dentro de las amebas, lo que les permite sobrevivir y prosperar dentro de otras células fagocíticas (por ejemplo, macrófagos humanos). La tenacidad de AVL para proliferar en ecosistemas hostiles y sobrevivir a la desecación o desinfección hace de estos organismos una fuente potencial transmitida por el agua, por los alimentos y el aire. Conocidos por su ubicuidad, los AVL han sido aislados de diversas matrices ambientales, como suelo, agua dulce y salobre, aguas residuales, aguas termales, polvo y otros. En función de lo antes comentado, este trabajo tiene como objetivo identificar amebas de vida libre en tanques de almacenamiento de agua de edificios del área metropolitana de la ciudad de Ambato (Ecuador), con el fin de aplicar soluciones correctivas en cualquiera de sus puntos. Según el sitio de muestreo, el género de AVL identificado con mayor frecuencia en la superficie fue Balamuthia sp (70,59%) y Acanthamoeba sp., con un 37,78 %. El fondo estuvo caracterizado por Naegleria sp. con un 44,44%(AU)

Naegleria fowleri, Acanthamoeba spp. and Balamuthia mandrillaris are species of free-living amoeba (FLA) that cause severe central nervous system disease and can be fatal in humans and other animals. All genera of FLA have two stages: cyst and trophozoite. Trophozoites divide by binary fission, actively feed, and is the infective stage of the organism. In the environment, acanthopods allow trophozoites to adhere to the surfaces of bacteria, fungi, algae, and detritus on which their food is found. Acanthamoeba spp. they appear to maintain an endosymbiotic relationship with some of the bacteria they consume. Many of these bacteria, such as Legionella, Pseudomonas, Vibrio, and some mycobacteria appear to acquire virulence factors while living inside amoebas, allowing them to survive and thrive inside other phagocytic cells (eg, human macrophages). FLA's tenacity to proliferate in hostile ecosystems and survive desiccation or disinfection makes these organisms a potential waterborne, foodborne, and airborne source. Known for their ubiquity, FLAs have been isolated from various environmental matrices, such as soil, fresh and brackish water, wastewater, hot springs, dust, and others. Based on the aforementioned, this work aims to identify free-living amoebas in water storage tanks of buildings in the metropolitan area of the city of Ambato (Ecuador), in order to apply corrective solutions at any of its points. According to the sampling site, the FLA genus most frequently identified on the surface was Balamuthia sp (70.59%) and Acanthamoeba sp., with 37.78 %. The background was characterized by Naegleria sp. with 44.44%(AU)

The AraC-type transcription factor TagK is a new player in the signaling cascade that induces the anti-eukaryotic T6SS of Xanthomonas citri.

Type 6 secretion systems (T6SSs) are specialized multiprotein complexes that inject protein effectors into prokaryotic and/or eukaryotic cells. We previously described the role of the T6SS of the phytopathogen Xanthomonas citri pv. citri as an anti-eukaryotic nanoweapon that confers resistance to predation by the amoeba Dictyostelium discoideum. Transcription of the X. citri T6SS genes is induced by a signaling cascade involving the Ser/Thr kinase PknS and the extracytoplasmic function sigma factor EcfK. Here, we used a strain overexpressing a phosphomimetic constitutively active version of EcfK (EcfKT51E ) to identify the EcfK regulon, which includes a previously uncharacterized transcription factor of the AraC-family (TagK), in addition to T6SS genes and genes encoding protein homeostasis factors. Functional studies demonstrated that TagK acts downstream of EcfK, binding directly to T6SS gene promoters and inducing T6SS expression in response to contact with amoeba cells. TagK controls a small regulon, consisting of the complete T6SS, its accessory genes and additional genes encoded within the T6SS cluster. We conclude that a singular regulatory circuit consisting of a transmembrane kinase (PknS), an alternative sigma factor (EcfK) and an AraC-type transcriptional regulator (TagK) promotes expression of the X. citri T6SS in response to a protozoan predator.

Anaerobic ATP synthesis pathways and inorganic phosphate transport and their possible roles in encystment in Acanthamoeba castellanii.

Acanthamoeba castellanii is the etiological agent of amoebic keratitis and is present in the environment in trophozoite or cyst forms. Both forms can infect the vertebrate host and colonize different tissues. The high resistance of cysts to standard drugs used in clinics contributes to the lack of effective treatments. Therefore, in this context, studies have emerged to understand cyst physiology and metabolism. Phosphate transporters are proteins responsible for the uptake of extracellular inorganic phosphate and transport to the cytosol. This work aims to verify the relationship between Pi transport and energetic metabolism in cysts of A. castellanii. The phosphate uptake ratio was higher in cysts compared with trophozoites. Recently, three sequences related to phosphate transporters have been identified in the A. castellanii genome (AcPHS1, AcPHS2, and AcPHS3); the messenger RNA expression levels of which differ depending on the amoeba life form. Pi uptake in cysts displayed peak activity at alkaline pH, whereas Pi transport in trophozoites was not affected in the same pH ranges. Cysts harbor a low-affinity Pi transport system (K0,5 and Vmax values of 1.76 ± 0.26 mM and 104.6 ± 6.3 nmol Pi × h-1 × 106 cells) compared to the trophozoite phosphate transport system. Pi transport seems important for anaerobic adenosine triphosphate synthesis in cysts, which initially occurs through the glycolytic pathway and subsequently through the pyruvate ferredoxin oxidoreductase pathway. Altogether, these results suggest that contrary to that previously postulated, cysts are active metabolic forms, and, as noted in trophozoites, phosphate uptake is important for energetic metabolism.

Cutaneous balamuthiasis: A clinicopathological study.

INTRODUCTION: Balamuthia mandrillaris, a free-living amoeba, causes an uncommon infection that is characterized by cutaneous and neurological involvement, which carries a poor prognosis. METHODS: This is a retrospective observational study including patients with clinical suspicion of cutaneous balamuthiasis, their skin biopsies, and/or a positive direct immunofluorescence test. The data were collected from the Dermatology and Pathology service of the Hospital Cayetano Heredia and the Instituto Tropical Alexander von Humboldt, Lima, Peru, from January 1985 to June 2007. We identified 60 biopsies from 35 patients, from which clinical data were available in 30. RESULTS: Twenty-two (73%) patients had centrofacial lesions, mostly located on the nose. The classical lesion was an asymptomatic, erythematous, or violaceous infiltrated plaque. Twenty-two (73%) patients had neurologic involvement. Fifty (83%) biopsies showed granulomatous dermatitis and 75% showed ill-defined tuberculoid granulomas without caseous necrosis. Multinucleated giant cells were observed in 52 (87%) biopsies. Trophozoite forms were identified in the biopsies of 25 (71%) patients. Direct immunofluorescence was positive in 25 (71%) patients. CONCLUSION: B. mandrillaris is a pathogen that is capable of inducing a characteristic skin lesion with a reaction pattern of ill-defined tuberculoid granulomas and many giant cells.

Nisin Induces Cell-Cycle Arrest in Free-Living Amoebae Acanthamoeba castellanii.

PURPOSE: Acanthamoeba spp. are free-living amoebas with worldwide distribution and play an important role as disease-causing agents in humans. Drug inability to completely eradicate these parasites along with their toxic effects suggest urgent need for new antimicrobials. Nisin is a natural antimicrobial peptide produced by Lactococcus lactis. Nisin is also the only bacteriocin approved for use in food preservation. In this work, we analyzed the effect of nisin on the growth of Acanthamoeba castellanii trophozoites. METHODS: A total of 8 × 104 trophozoites were exposed to increasing concentrations of nisin to determine its activity. Changes in cell membrane and cellular cycle of trophozoites were investigated by flow cytometry, and nisin cytotoxicity in mammalian cells was evaluated in L929 cells by MTT method. RESULTS: After 24 h exposure to increasing nisin concentrations, an IC50 of 4493.2 IU mL-1 was obtained for A. castellanii trophozoites. However, after 72 h a recovery in amoebic growth was observed, and it was no longer possible to determine IC50. Flow cytometry analysis showed that nisin has no effect on the membrane integrity. Treatment with nisin induced cell-cycle arrest during G1 and S phases in A. castellanii trophozoites, which recovered their growth after 72 h. CONCLUSION: This is one of the first studies showing the effect of internationally approved nisin against A. castellanii trophozoites. Nisin caused cell-cycle arrest in trophozoites, momentarily interfering with the DNA replication process. The data highlight the amoebostatic activity of nisin, and suggest its use as an adjuvant for the treatment of infections caused by Acanthamoeba spp.

A history of over 40 years of potentially pathogenic free-living amoeba studies in Brazil - a systematic review

Free-living amoeba (FLA) group includes the potentially pathogenic genera Acanthamoeba, Naegleria, Balamuthia, Sappinia, and Vermamoeba, causative agents of human infections (encephalitis, keratitis, and disseminated diseases). In Brazil, the first report on pathogenic FLA was published in the 70s and showed meningoencephalitis caused by Naegleria spp. FLA studies are emerging, but no literature review is available to investigate this trend in Brazil critically. Thus, the present work aims to integrate and discuss these data. Scopus, PubMed, and Web of Science were searched, retrieving studies from 1974 to 2020. The screening process resulted in 178 papers, which were clustered into core and auxiliary classes and sorted into five categories: wet-bench studies, dry-bench studies, clinical reports, environmental identifications, and literature reviews. The papers dating from the last ten years account for 75% (134/178) of the total publications, indicating the FLA topic has gained Brazilian interest. Moreover, 81% (144/178) address Acanthamoeba-related matter, revealing this genus as the most prevalent in all categories. Brazil's Southeast, South, and Midwest geographic regions accounted for 96% (171/178) of the publications studied in the present work. To the best of our knowledge, this review is the pioneer in summarising the FLA research history in Brazil.

Primary Amoebic Meningoencephalitis by Naegleria fowleri: Pathogenesis and Treatments.

Naegleria fowleri is a free-living amoeba (FLA) that is commonly known as the "brain-eating amoeba." This parasite can invade the central nervous system (CNS), causing an acute and fulminating infection known as primary amoebic meningoencephalitis (PAM). Even though PAM is characterized by low morbidity, it has shown a mortality rate of 98%, usually causing death in less than two weeks after the initial exposure. This review summarizes the most recent information about N. fowleri, its pathogenic molecular mechanisms, and the neuropathological processes implicated. Additionally, this review includes the main therapeutic strategies described in case reports and preclinical studies, including the possible use of immunomodulatory agents to decrease neurological damage.

Encefalitis por ameba de vida libre: reporte de caso / Free-living amoeba encephalitis: case report

La encefalitis amebiana granulomatosa es una enfermedad considerada atípica debido a la baja incidencia desde que fue descubierta. Sin embargo, conforme han avanzando los métodos diagnósticos, en retrospectiva, se ha descrito la presencia de esta enfermedad desde principios del siglo XX, identificando a Acanthamoeba spp y Balamuthia mandrillaris como agentes más comunes.Es una enfermedad de difícil diagnóstico debido al amplio abanico clínico que puede presentar, siendo el sitio anatómico dentro del sistema nervioso central quien determinará estas características. El conocimiento y sospecha de la misma permitirá un abordaje terapéutico oportuno aunque, en la mayoría de los casos, con un desenlace fatal para el paciente.A continuación se presenta el caso clínico de un paciente masculino de 26 años de edad originario de la ciudad de Durango, México, sin antecedentes de importancia, de ocupación ladrillero, quien sufre golpe con ladrillo en región naso-orbitaria, ingresando al hospital al tercer día del evento con diagnóstico de celulitis periorbitaria izquierda y nasal, cefalea de leve intensidad, rinorrea abundante, hemiparesia fasciocorporal izquierda, parestesia de extremidad superior izquierda, diplopía y vértigo postural. Los estudios imagenológicos revelaron lesiones heterogéneas en el lóbulo occipital, hemisferio cerebeloso izquierdo y tallo cerebral. Se realizó escisión de lesión tumoral cerebelar, con estudio anatomopatológico, reportando encefalitis granulomatosa por presencia de ameba de vida libre. El paciente evolucionó hacia deterioro rostro-caudal declarándose su fallecimiento a los 16 días de su estancia intrahospitalaria.En el Estado de Durango, México, no se cuenta con casos reportados por infección de ameba de vida libre

Granulomatous amoebic encephalitis is a disease considered atypical due to the low incidence since it was discovered. However, as diagnostic methods have progressed, in retrospect, the presence of this disease has been described since the beginning of the 20th century, identifying Acanthamoeba spp. and Balamuthia mandrillaris as the most common agents.A disease that is difficult to diagnose due to the wide clinical range that it can present, being the anatomical site within the central nervous system that will determine these characteristics. The knowledge and suspicion of it will allow a timely therapeutic approach although, in most cases, with a fatal outcome for the patient.The clinical case is presented below of a 26 year old male patient from the city of Durang, México, with no significant history, of brick occupation, who suffers a blow with a brick in the naso-orbital region, follows, entering our hospital on the third day of the event with a diagnosis of left periorbital and nasal cellulitis, mild headache, profuse rhinorrhea, left fasciocorporeal hemiparesis, left upper limb parestesia, diplopia, and postural vertigo.Imaging studies were performed observing heterogeneous lesions in the occipital lobe, left cerebellar hemisphere and brain stem; excision of the cerebellar tumor lesion being carried out, with pathological study, reporting granulomatous encephalitis due to the presence of free-living amoeba. The patient progresses to face-caudal deterioration, declaring his death after 16 days of hospital stay.Within the state of Durango, Mexico, there are no reported cases of free-living amoeba infection

Amoebal coculture and enrichment methods as a proposal for water quality control in Brazil.

Free living amoebae (FLA) can be found in different environments, where they feed on diverse microorganisms. Some bacteria preyed by FLA are called amoeba-resistant bacteria (ARB), as they can resist to lysosomal fusion and are capable of multiplying and evading FLA after internalization, propagating in the environment. Despite the health risks due to the existence of pathogenic and opportunistic species that are ARB and the pathogenicity of some FLA species, there are no water quality protocols to analyze the presence of ARB or FLA. In this sense, our study aimed to isolate FLA through amoebal enrichment and to identify ARB using amoebal coculture in water samples from a public park and two hospitals in southern Brazil. As a result, 9 different microorganisms genera have been identified through amoebal coculture, including fastidious Legionella spp. and Bosea vestrisii. From the positive samples for FLA, by amoebal enrichment, Acanthamoeba spp., Vermamoeba vermiformis and Naegleria spp. were identified in 14 amoebic isolates. The methodologies used in this work proved to be effective as simple and low-cost methods to be used in the implementation in water quality control of anthropogenic environments.

Single cell cloning of free-living amoebae with isolated square agar.

An alternative amoeba cloning technique is described. The amoebic cells were scraped from the agar and inoculated in a new agar medium separated in isolated squares, preventing a cell from invading the space of another cell, increasing the reliability of cloning, and providing an efficient quality control of technique.

The role of Acanthamoeba spp. in biofilm communities: a systematic review.

Acanthamoeba spp. have always caused disease in immunosuppressed patients, but since 1986, they have become a worldwide public health issue by causing infection in healthy contact lens wearers. Amoebae of the Acanthamoeba genus are broadly distributed in nature, living either freely or as parasites, and are frequently associated with biofilms throughout the environment. These biofilms provide the parasite with protection against external aggression, thus favoring its increased pathogeny. This review aims to assess observational studies on the association between Acanthamoeba spp. and biofilms, opening potential lines of research on this severe ocular infection. A systematic literature search was conducted in May 2020 in the following databases: PubMed Central®/Medline, LILACS, The Cochrane Library, and EMBASE®. The studies were selected following the inclusion and exclusion criteria specifically defined for this review. Electronic research recovered 353 publications in the literature. However, none of the studies met the inclusion criterion of biofilm-producing Acanthamoeba spp., inferring that the parasite does not produce biofilms. Nonetheless, 78 studies were classified as potentially included regarding any association of Acanthamoeba spp. and biofilms. These studies were allocated across six different locations (hospital, aquatic, ophthalmic and dental environments, biofilms produced by bacteria, and other places). Acanthamoeba species use biofilms produced by other microorganisms for their benefit, in addition to them providing protection to and facilitating the dissemination of pathogens residing in them.

Primer informe clínico de Balamuthia mandrillaris en el distrito de Camaná, Arequipa, Perú / First case report of the Balamuthia mandrillaris in the Camaná district of Arequipa, Peru

Resumen Las infecciones por amebas de vida libre causan compromiso cutáneo y neurológico. Estas afecciones presentan una baja frecuencia pero una alta letalidad, mayor al 98%. Gene ralmente el cuadro clínico es inespecífico y los exámenes de laboratorio no son de gran ayuda, por lo que representa un reto diagnóstico y terapéutico. En este informe presentamos el caso de un paciente de 21 años que fue hospitalizado por un síndrome convulsivo con tumoraciones cerebrales, además de una lesión cutánea en el muslo derecho. El análisis histopatológico, PCR y el cuadro clínico permitieron el diagnóstico de encefalitis amebiana granulomatosa. A pesar del tratamiento, el paciente tuvo un desenlace fatal.

Abstract Infections by free-living amoebas cause cutaneous and neurological compromise. These conditions have a low frequency, but a high lethality more than 98%. Generally, the clinical picture is nonspecific; the laboratory tests dont help, so it represents a diagnostic and therapeutic challenge. In this report, we present the case of a 21-year-old patient, who was hospitalized for a convulsive syndrome with brain tumors, in addition to a cutaneous lesion on the right thigh. Histopathological analysis, PCR and the clinical picture allowed the diagnosis of granulomatous amebic encephalitis. Despite the treatment, the patient had a fatal outcome.

Transcriptional Control of the Production of Aspergillus fumigatus Conidia-Borne Secondary Metabolite Fumiquinazoline C Important for Phagocytosis Protection.

Aspergillus fumigatus produces diverse secondary metabolites whose biological functions and regulation remain to be understood. Despite the importance of the conidia for this fungus, the role of the conidia-born metabolite fumiquinazoline C (FqC) is unclear. Here, we describe a dual function of the cell-wall integrity pathway in regulating FqC biosynthesis dictated by the MAPK kinase MpkA, which phosphorylates one of the nonribosomal peptide synthetases enzymes of the cluster (FmqC), and the transcription factor RlmA, which directly regulates the expression of fmq genes. Another level of crosstalk between the FqC regulation and the cell physiology is described since the deletion of the stress-responsive transcription factor sebA provokes derepression of the fmq cluster and overproduction of FqC. Thus, we describe a mechanism by which A. fumigatus controls FqC biosynthesis orchestrated by MpkA-RlmA and SebA and hence enabling survival and adaptation to the environmental niche, given that FqC is a deterrent of ameba predation.

[First case report of the Balamuthia mandrillaris in the Camaná district of Arequipa, Peru]. / Primer informe clínico de Balamuthia mandrillaris en el distrito de Camaná, Arequipa, Perú.

Infections by free-living amoebas cause cutaneous and neurological compromise. These conditions have a low frequency, but a high lethality more than 98%. Generally, the clinical picture is nonspecific; the laboratory tests dont help, so it represents a diagnostic and therapeutic challenge. In this report, we present the case of a 21-year-old patient, who was hospitalized for a convulsive syndrome with brain tumors, in addition to a cutaneous lesion on the right thigh. Histopathological analysis, PCR and the clinical picture allowed the diagnosis of granulomatous amebic encephalitis. Despite the treatment, the patient had a fatal outcome.

Presence and interaction of free-living amoebae and amoeba-resisting bacteria in water from drinking water treatment plants.

Free-living amoebae (FLA) are ubiquitous and many isolates have been shown to be infected with amoeba-resisting bacteria, as the example of Acanthamoeba and Legionella interaction. Due to the high environmental prevalence of Acanthamoeba. in the Castilian Plateau (Spain), the aims of this work were to investigate the occurrence of Acanthamoeba and other FLA in water from several sampling points from four Drinking Water Treatment Plants (DWTP) and to investigate the presence of Legionella spp. and other amoeba-resisting bacteria in biofilms in raw and finished water, taking into account that no legislation exists for this protozoa control. Acanthamoeba was detected at different sampling points, and sand filters seemed to contribute to amoebic enrichment. After ozonation, a temporary decrease in viable amoebae was observed. The genotypes detected were T3, T4, and T5, revealing the first report of genotype T5 in waters from this region. Moreover, Balamuthia mandrillaris, Vermamoeba vermiformis and Paravahlkampfia sp. were detected. Regarding Legionella, PCR detection in raw and finished water was higher than by agar culture, but even higher after Acanthamoeba co-culture. Also, Legionella's presence was higher in raw water than in finished water. The decrease of free Legionella observed from raw (27.5%, by PCR) to finished water (3.4% by PCR) contrasted with the increase of Legionella-infected FLA from raw (30.7%) to finished water (52%). At biofilm, free Legionella was not detected, and the percentage of infected FLA was low (3.8%). Legionella species identified in these samples were L. drozanskii, L. donaldsonii and L. feeleii. Additionally, Acanthamoeba co-culture led to the isolation of Pseudomonas aeruginosa, P. stutzeri, P. fluorecens, Achromobacter xylosoxidans and Stenotrophomonas maltophilia. The highly disseminated presence of Acanthamoeba and the detection of amoeba-resisting bacteria inside amoebae highlight the importance of developing methods for controlling FLA in order to limit human pathogenic amoeba-resisting bacteria survival to the water purification processes.