Arcobacter butzleri survives within trophozoite of Acanthamoeba castellanii

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The plasma EBV load showed high specificity to detect all PTLD; while an increasing sensitivity was noted as PTLD stages progressed. Total PTLD: included categories 1–4 (A); advanced PTLD: categories 2–4 (B)." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "María Dolores Fellner, Karina A. Durand, Veronica Solernou, Andrea Bosaleh, Ziomara Balbarrey, María T. García de Dávila, Marcelo Rodríguez, Lucía Irazu, Lidia V. Alonio, María A. Picconi" "autores" => array:10 [ 0 => array:2 [ "nombre" => "María Dolores" "apellidos" => "Fellner" ] 1 => array:2 [ "nombre" => "Karina A." "apellidos" => "Durand" ] 2 => array:2 [ "nombre" => "Veronica" "apellidos" => "Solernou" ] 3 => array:2 [ "nombre" => "Andrea" "apellidos" => "Bosaleh" ] 4 => array:2 [ "nombre" => "Ziomara" "apellidos" => "Balbarrey" ] 5 => array:2 [ "nombre" => "María T." "apellidos" => "García de Dávila" ] 6 => array:2 [ "nombre" => "Marcelo" "apellidos" => "Rodríguez" ] 7 => array:2 [ "nombre" => "Lucía" "apellidos" => "Irazu" ] 8 => array:2 [ "nombre" => "Lidia V." "apellidos" => "Alonio" ] 9 => array:2 [ "nombre" => "María A." "apellidos" => "Picconi" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0325754116000110?idApp=UINPBA00004N" "url" => "/03257541/0000004800000002/v1_201607030039/S0325754116000110/v1_201607030039/en/main.assets" ] "itemAnterior" => array:19 [ "pii" => "S0325754116000079" "issn" => "03257541" "doi" => "10.1016/j.ram.2016.02.002" "estado" => "S300" "fechaPublicacion" => "2016-04-01" "aid" => "88" "copyright" => "Asociación Argentina de Microbiología" "documento" => "article" "crossmark" => 1 "licencia" => "http://creativecommons.org/licenses/by-nc-nd/4.0/" "subdocumento" => "fla" "cita" => "Rev Argent Microbiol. 2016;48:101-4" "abierto" => array:3 [ "ES" => true "ES2" => true "LATM" => true ] "gratuito" => true "lecturas" => array:2 [ "total" => 1375 "formatos" => array:3 [ "EPUB" => 39 "HTML" => 814 "PDF" => 522 ] ] "es" => array:12 [ "idiomaDefecto" => true "cabecera" => "INFORME BREVE" "titulo" => "Evaluación de la transmisión galactógena de Trichinella patagoniensis en ratones BALB/c" "tienePdf" => "es" "tieneTextoCompleto" => "es" "tieneResumen" => array:2 [ 0 => "es" 1 => "en" ] "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "101" "paginaFinal" => "104" ] ] "titulosAlternativos" => array:1 [ "en" => array:1 [ "titulo" => "Assessment of transmammary transmission of Trichinella patagoniensis in BALB/c mice" ] ] "contieneResumen" => array:2 [ "es" => true "en" => true ] "contieneTextoCompleto" => array:1 [ "es" => true ] "contienePdf" => array:1 [ "es" => true ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Fernando A. Fariña, Mariana I. Pasqualetti, Natalia M. Cardillo, Tatiana Aronowicz, Mariano Ercole, Silvio J. Krivokapich, Mabel M. Ribicich" "autores" => array:7 [ 0 => array:2 [ "nombre" => "Fernando A." "apellidos" => "Fariña" ] 1 => array:2 [ "nombre" => "Mariana I." "apellidos" => "Pasqualetti" ] 2 => array:2 [ "nombre" => "Natalia M." "apellidos" => "Cardillo" ] 3 => array:2 [ "nombre" => "Tatiana" "apellidos" => "Aronowicz" ] 4 => array:2 [ "nombre" => "Mariano" "apellidos" => "Ercole" ] 5 => array:2 [ "nombre" => "Silvio J." "apellidos" => "Krivokapich" ] 6 => array:2 [ "nombre" => "Mabel M." "apellidos" => "Ribicich" ] ] ] ] ] "idiomaDefecto" => "es" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0325754116000079?idApp=UINPBA00004N" "url" => "/03257541/0000004800000002/v1_201607030039/S0325754116000079/v1_201607030039/es/main.assets" ] "en" => array:20 [ "idiomaDefecto" => true "cabecera" => "Brief reports" "titulo" => "Arcobacter butzleri survives within trophozoite of Acanthamoeba castellanii" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "105" "paginaFinal" => "109" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "María P. Villanueva, Gustavo Medina, Heriberto Fernández" "autores" => array:3 [ 0 => array:3 [ "nombre" => "María P." "apellidos" => "Villanueva" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 1 => array:3 [ "nombre" => "Gustavo" "apellidos" => "Medina" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] 2 => array:4 [ "nombre" => "Heriberto" "apellidos" => "Fernández" "email" => array:1 [ 0 => "hfernand@uch.cl" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "*" "identificador" => "cor0005" ] ] ] ] "afiliaciones" => array:2 [ 0 => array:3 [ "entidad" => "Instituto de Microbiología Clínica, Universidad Austral de Chile, PO Box 567, Valdivia, Chile" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Facultad de Ciencias de la Salud, Universidad Católica de Temuco, PO Box 15-D, Temuco, Chile" "etiqueta" => "b" "identificador" => "aff0010" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Arcobacter butzleri sobrevive en el interior de trofozoitos de Acanthamoeba castellanii" ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1009 "Ancho" => 2040 "Tamanyo" => 261224 ] ] "descripcion" => array:1 [ "en" => "Phase contrast microscopy. (A) Trophozoite of A. castellanii showing A. butzleri seen in close association with A. castellanii gathering at one pole of the amoeba cell known as food-cup (30min of co-incubation). (B) Trophozoite of A. castellanii showing A. butzleri within the amoebic vacuoles (50min of co-incubation)." ] ] ] "textoCompleto" => "The genus Arcobacter was originally proposed by Vandamme et al.<a class="elsevierStyleCrossRef" href="#bib0145">14</a> and includes bacteria previously considered as members of the genus Campylobacter. Today, the genus Arcobacter is represented by 21 species isolated from human, animals and environmental samples<a class="elsevierStyleCrossRef" href="#bib0150">15</a>. However, only four species have been isolated from human and animal samples and they are considered of medical interest: Arcobacter cryaerophilus, Arcobacter butzleri, Arcobacter skirrowii and Arcobacter thereius<a class="elsevierStyleCrossRef" href="#bib0140">13</a>.A. butzleri is the species that is most frequently isolated from environmental water, food and clinical samples. From a public health point of view, it is considered an emerging foodborne and zoonotic enteropathogen transmitted by food and water. Furthermore, it has been ranked as the fourth most common member of Campylobacteraceae isolated from human feces<a class="elsevierStyleCrossRefs" href="#bib0085">2,5,13</a>. Thus, it has become increasingly important as an agent of diarrhea in human beings, recognizing as potential routes of infection the consumption and manipulation of contaminated raw or poorly cooked food of animal origin and untreated water<a class="elsevierStyleCrossRefs" href="#bib0085">2,5,13</a>.One of the main reservoirs of this species is water in diverse environments<a class="elsevierStyleCrossRef" href="#bib0085">2</a> where they may interact with other microorganisms, which are natural inhabitants of hydric ecosystems, in particular with free-living amoebas.Free-living amoebas of the Genus Acanthamoeba are widely distributed in water environments. Amoebic trophozoites are part of the indigenous inhabitants of natural water bodies, sharing this habitat with other microorganisms, mainly bacteria. Since these bacteria can survive for diverse periods within the amoeba, many researchers have suggested that these protozoa may be a reservoir for bacterial species<a class="elsevierStyleCrossRef" href="#bib0080">1</a>. Because of the particular ability of some bacteria to survive within amoebae, the members of genus Acanthamoeba could play a role in the transmission of some enteric bacteria. The survival of Campylobacter jejuni inside Acanthamoeba and its transmission to chicken as an endosymbiont of A. castellanii and Acanthamoeba polyphaga<a class="elsevierStyleCrossRef" href="#bib0080">1</a> has been demonstrated.Since A. butzleri is a bacterium that can be recovered from water environments and food products, such as mussels produced in estuarine environments<a class="elsevierStyleCrossRefs" href="#bib0085">2,6</a>, it could share this environment with free-living amoebas, probably establishing close relationships with these protozoa, similarly to those described in the genus Campylobacter<a class="elsevierStyleCrossRefs" href="#bib0080">1,11,12</a>.In order to demonstrate if A. butzleri is capable of establishing similar close relations with free-living amoebas, we determined the survival capacity of Arcobacter inside A. castellanii.Three strains of A. butzleri were used for this study. Strain F215 (isolated from child human stools), strain Puar190 (isolated from chicken meat) and the reference strain LMG10828 (isolated from human feces) used as control strain and kindly provided by Dr. Peter Vandamme (Laboratory of Microbiology, Ghent University, Belgium). These strains were stored at −80°C in brain-heart infusion broth supplemented with 15% glycerol. Before each experiment, bacteria were grown on blood agar plates incubated at 26°C for 48h under aerobic conditions.A. castellanii strain BP91/2760 was originally isolated from the eye of a patient with keratitis and was kindly provided by Dr. Winiecka-Krusnell (Public Health Agency of Sweden). Amoebae were maintained axenically in peptone yeast extract glucose (PYG) medium at 26°C as monolayers in 25cm2 flasks and were examined under an inverted microscope before use. To assess the number of amoebae, counts were made using a Neubauer chamber.To establish A. butzleri/A. castellanii early relationships, A. castellanii were grown to confluence in 25cm2 flasks with 5ml of PYG medium. The flask were gently shaken and the PYG with non-adherent amoebae was removed and replaced with fresh medium; then the amoebae were incubated on ice for 30min to weaken adherence. The suspensions were centrifuged at 203g for 10min and the pellet was washed three times with Page's saline solution. After the last centrifugation, the pellet was suspended in PYG and the total number of amoebae was estimated. Aliquots of 1ml with approximately 6×104 trophozoites were placed in each well of a 24-well tissue culture dish. Bacterial strains suspended in Page's saline solution to a density of approximately 1.2×108 colony forming units (CFU), and 100ml of this suspension was added to each well containing the amoebae and the plate was incubated at 26°C under aerobic conditions to induce the infection of the amoebas by Arcobacter strains in a 1:2000 ratio. As bacterial survival controls, three wells were inoculated with A. butzleri suspended in Page's saline solution without amoebae. Finally, the assessment of the early interactions between bacteria and amoebae was performed by taking aliquots of the suspension at 30 and 50min and observed directly under phase contrast microscope.Long-term interaction between bacteria and amoebae was studied at 24, 48, 72, 120, 168, and 240h using a modification of the gentamicin protection assay proposed by Dirks and Quinlan<a class="elsevierStyleCrossRef" href="#bib0090">3</a>. In brief, after the early interaction assay, 100μg/ml gentamicin were added to each well and incubated at room temperature for 1h to kill extra-amoebic bacterial cells. Amoebas were harvested from the well and washed three times with Page's saline solution. After the third wash, an aliquot of 100μl was taken and plated on blood agar plates to determine the number of extracellular bacteria that might have escaped the gentamicin treatment at each sampling time. 100μl of 0.5% sodium desoxycholate was added to another aliquot for 25–30min to perform the amoeba lysis. An aliquot of 100μl of the lysate suspension was taken to count the number of total bacteria by CFU counts (intracellular+extracellular bacteria). Abundance of viable intracellular bacteria was determined by subtracting the number of extracellular bacteria from the total count. All these experiments were carried out in triplicate. The amoebae/bacteria interaction was also monitored and documented through phase contrast microscopy and by bright-field microscopy.The results of the assays were divided into two groups in accordance with the duration of the tests performed.A. – Early interactions between A. butzleri and A. castellanii. In the first 30min of co-incubation, A. butzleri could be seen in close association with A. castellanii, at this time most of the bacteria showed a tendency to gather at one pole of the amoeba cell (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>A). Soon after 50min of co-culture, A. butzleri cells were detected inside the amoeba. The internalized A. butzleri were located in vacuoles and never in the amoeba cytoplasm (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>B). These observations are consistent with our previous studies<a class="elsevierStyleCrossRefs" href="#bib0110">7,8</a>, confirming an initial contact pattern in the interaction A. butzleri/A. castellanii.<elsevierMultimedia ident="fig0005"></elsevierMultimedia>The internalization of A. butzleri seems to be an active and probable metabolic process since all intracellular bacteria are located in vacuoles and never inside the amoeba cytoplasmic region. These images are similar to those previously reported by our group<a class="elsevierStyleCrossRefs" href="#bib0110">7,8</a>.This early observation indicated that the interaction between bacteria and amoebae could be a common phenomenon in Arcobacter, which is time-efficient. Recently, Medina et al.<a class="elsevierStyleCrossRef" href="#bib0115">8</a> have reported that the attachment process of A. butzleri to A. castellanii involves the participation of mannose-binding proteins and membrane-associated receptors of glucose and galactose present in the amoebae, whereas in their internalization, the protozoan actin polymerization plays an active role.B. – Time course of the interaction of A. butzleri and A. castellanii. The long-term experiments demonstrated that A. butzleri strains F-215 and PUAr190 were able to survive for at least 240h inside the amoebas. On the other hand, control assays done to establish whether the presence of A. butzleri could affect the survival of the amoebae, showed no significant differences in the survival rates of the amoebae with and without bacteria (data not shown).Regarding the survival of A. butzleri within A. castellanii trophozoites at different time points, all the bacteria studied in the co-culture with the amoeba as well as control bacteria showed a decrease in CFU values until reaching the minimum count on the tenth day. There is a clear difference in the survival of these three strains of A. butzleri inside the amoebae. The data suggest that recently isolated A. butzleri strains F-215 and PUAr190 have better survival than the reference strain LMG10828. Their colony counts suggest that this strain does not multiply inside A. castellani, not reaching the high numbers seen for the other two strains (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>). Furthermore, their UFC numbers drop in a similar way as the number of viable bacterial cells decays in the control assay without amoebae. Whether those differences are related to the source of isolation, geographic origin or number of subcultures is unknown, requiring further investigations.<elsevierMultimedia ident="fig0010"></elsevierMultimedia>The methodology used in this study to estimate the intra-amoeba bacterial survival rate was selected because other methods, such as vital stains and counting by microscopy, express their results as percentages and do not allow to determine the bacterial number as accurately as the gentamicin protection assay does<a class="elsevierStyleCrossRef" href="#bib0090">3</a>.The A. butzleri strains used in this study were not only able to infect A. castellanii but also capable of surviving inside the vacuoles, in which we confirmed that bacteria were actively motile. Through the phagolysosome formation analysis, Medina et al.<a class="elsevierStyleCrossRef" href="#bib0115">8</a> concluded that the survival of A. butzleri inside the amoebae could be related to their ability to remain inside vacuoles not fused with lysosomes, or with their ability to retard phagolysosome formation. However, further investigations are needed to elucidate the survival mechanisms of A. butzleri inside free-living amoebae.Transmissibility of enteropathogens, particularly those associated in zoonosis, has been difficult to identify and for the same reason, nearly impossible to prevent. In the last quarter of the 20th century and the beginning of this century, emerging pathogens have become a major health problem<a class="elsevierStyleCrossRef" href="#bib0095">4</a>. Recent observations showed that free-living amoebae may act as a reservoir and vehicle for the survival and transmission of C. jejuni and other enteropathogens, providing a unique form of self-perpetuation of these pathogens<a class="elsevierStyleCrossRefs" href="#bib0080">1,9,10,12</a>.Our team has been very interested in the role of Arcobacter species as human pathogens and its association with animal sources<a class="elsevierStyleCrossRefs" href="#bib0100">5,6</a>. Different studies have indicated that A. butzleri can survive under several environmental conditions and can be isolated from diverse hosts like farm animals, seafood, and hose pets<a class="elsevierStyleCrossRefs" href="#bib0085">2,5,6</a>. In the present study, the survival of A. butzleri inside A. castellanii strongly suggests that this protozoon can serve as a reservoir for A. butzleri and it might be an important vehicle in the transmission of this human emergent zoonotic pathogen, acting as a Trojan horse<a class="elsevierStyleCrossRefs" href="#bib0085">2,8,12</a>. Experimental transmissibility assays of A. butzleri inside A. castellanii into mice, currently performed in our laboratory (data not shown), allow to infer the eventual participation of this protozoon as transmission vehicle of A. butzleri. Further studies have to be carried out to elucidate this epidemiological angle of the A. butzleri/free-living amoebae relationships.Ethical disclosuresProtection of human and animal subjectsThe authors declare that no experiments were performed on humans or animals for this study.Confidentiality of dataThe authors declare that no patient data appear in this article.Right to privacy and informed consentThe authors declare that no patient data appear in this article.Financial supportGrants DID-UACH S-2007-37 and FONDECYT 1110202.Conflict of interestsThe authors declare that they have no conflict of interests." "textoCompletoSecciones" => array:1 [ "secciones" => array:9 [ 0 => array:3 [ "identificador" => "xres680252" "titulo" => "Abstract" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0005" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec686103" "titulo" => "Keywords" ] 2 => array:3 [ "identificador" => "xres680253" "titulo" => "Resumen" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0010" ] ] ] 3 => array:2 [ "identificador" => "xpalclavsec686104" "titulo" => "Palabras clave" ] 4 => array:3 [ "identificador" => "sec0005" "titulo" => "Ethical disclosures" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0010" "titulo" => "Protection of human and animal subjects" ] 1 => array:2 [ "identificador" => "sec0015" "titulo" => "Confidentiality of data" ] 2 => array:2 [ "identificador" => "sec0020" "titulo" => "Right to privacy and informed consent" ] ] ] 5 => array:2 [ "identificador" => "sec0025" "titulo" => "Financial support" ] 6 => array:2 [ "identificador" => "sec0030" "titulo" => "Conflict of interests" ] 7 => array:2 [ "identificador" => "xack229393" "titulo" => "Acknowledgements" ] 8 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2015-03-02" "fechaAceptado" => "2015-12-01" "PalabrasClave" => array:2 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec686103" "palabras" => array:4 [ 0 => "Arcobacter butzleri" 1 => "Acanthamoeba castellanii" 2 => "Survival" 3 => "Environmental reservoir" ] ] ] "es" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Palabras clave" "identificador" => "xpalclavsec686104" "palabras" => array:4 [ 0 => "Arcobacter butzleri" 1 => "Acanthamoeba castellanii" 2 => "Sobrevida" 3 => "Reservorio ambiental" ] ] ] ] "tieneResumen" => true "resumen" => array:2 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "The survival of three Arcobacter butzleri strains inside Acanthamoeba castellanii was assessed using axenic cultures of A. castellanii that were inoculated with the tested strains and incubated at 26°C under aerobic conditions for 240h. The behavior of bacteria in contact with amoebae was monitored using phase contrast microscopy. The bacterial survival rate within amoebae was assessed through counting colony forming units, using the gentamicin protection assay. All A. butzleri strains were able to survive during 240h within the amoebae, thus suggesting that (i) A. butzleri resists the amoebic digestion processes at least for the analyzed time; (ii) that A. castellanii could serve as an environmental reservoir for this bacterium, probably acting as a transmission vehicle for A. butzleri." ] "es" => array:2 [ "titulo" => "Resumen" "resumen" => "Se determinó la sobrevida de 3 cepas de Arcobacter butzleri en el interior de trofozoítos de Acanthamoeba castellanii utilizando cultivos axénicos de la ameba, los que fueron inoculados con las cepas bacterianas e incubados a 26°C en aerobiosis durante 240h. La interacción bacterias/amebas fue monitorizada mediante microscopía de contraste de fase. Las tasas de sobrevida bacteriana en el interior de las amebas fueron establecidas a través del recuento de unidades formadoras de colonias mediante ensayos de protección con gentamicina. Todas las cepas de A. butzleri fueron capaces de sobrevivir en el interior de la ameba al menos durante 240h, lo que sugiere, por un lado, que A. butzleri puede resistir los procesos de digestión propios de la ameba, al menos por el lapso aquí analizado y, por el otro, que A. castellanii podría actuar como reservorio ambiental de esta bacteria y, probablemente, como vehículo de transmisión de A. butzleri." ] ] "multimedia" => array:2 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1009 "Ancho" => 2040 "Tamanyo" => 261224 ] ] "descripcion" => array:1 [ "en" => "Phase contrast microscopy. (A) Trophozoite of A. castellanii showing A. butzleri seen in close association with A. castellanii gathering at one pole of the amoeba cell known as food-cup (30min of co-incubation). (B) Trophozoite of A. castellanii showing A. butzleri within the amoebic vacuoles (50min of co-incubation)." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 951 "Ancho" => 1631 "Tamanyo" => 79758 ] ] "descripcion" => array:1 [ "en" => "Average colony-forming units (CFU) of intracellular A. butzleri recovered after co-culture with A. castellanii. 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Arcobacter butzleri sobrevive en el interior de trofozoitos de Acanthamoeba castellanii

María P. Villanuevaa, Gustavo Medinab, Heriberto Fernándeza,

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hfernand@uch.cl

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a Instituto de Microbiología Clínica, Universidad Austral de Chile, PO Box 567, Valdivia, Chile

b Facultad de Ciencias de la Salud, Universidad Católica de Temuco, PO Box 15-D, Temuco, Chile

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    "titulo" => "<span class="elsevierStyleItalic">Arcobacter butzleri</span> survives within trophozoite of <span class="elsevierStyleItalic">Acanthamoeba castellanii</span>"
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          "en" => "<p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Phase contrast microscopy&#46; &#40;A&#41; Trophozoite of <span class="elsevierStyleItalic">A&#46; castellanii</span> showing <span class="elsevierStyleItalic">A&#46; butzleri</span> seen in close association with <span class="elsevierStyleItalic">A&#46; castellanii</span> gathering at one pole of the amoeba cell known as food-cup &#40;30<span class="elsevierStyleHsp" style=""></span>min of co-incubation&#41;&#46; &#40;B&#41; Trophozoite of <span class="elsevierStyleItalic">A&#46; castellanii</span> showing <span class="elsevierStyleItalic">A&#46; butzleri</span> within the amoebic vacuoles &#40;50<span class="elsevierStyleHsp" style=""></span>min of co-incubation&#41;&#46;</p>"
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    "textoCompleto" => "<span class="elsevierStyleSections"><p id="par0005" class="elsevierStylePara elsevierViewall">The genus <span class="elsevierStyleItalic">Arcobacter</span> was originally proposed by Vandamme <span class="elsevierStyleItalic">et al&#46;</span><a class="elsevierStyleCrossRef" href="#bib0145"><span class="elsevierStyleSup">14</span></a> and includes bacteria previously considered as members of the genus <span class="elsevierStyleItalic">Campylobacter</span>&#46; Today&#44; the genus <span class="elsevierStyleItalic">Arcobacter</span> is represented by 21 species isolated from human&#44; animals and environmental samples<a class="elsevierStyleCrossRef" href="#bib0150"><span class="elsevierStyleSup">15</span></a>&#46; However&#44; only four species have been isolated from human and animal samples and they are considered of medical interest&#58; <span class="elsevierStyleItalic">Arcobacter cryaerophilus</span>&#44; <span class="elsevierStyleItalic">Arcobacter butzleri</span>&#44; <span class="elsevierStyleItalic">Arcobacter skirrowii</span> and <span class="elsevierStyleItalic">Arcobacter thereius</span><a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">13</span></a>&#46;</p><p id="par0010" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">A&#46; butzleri</span> is the species that is most frequently isolated from environmental water&#44; food and clinical samples&#46; From a public health point of view&#44; it is considered an emerging foodborne and zoonotic enteropathogen transmitted by food and water&#46; Furthermore&#44; it has been ranked as the fourth most common member of <span class="elsevierStyleItalic">Campylobacteraceae</span> isolated from human feces<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">2&#44;5&#44;13</span></a>&#46; Thus&#44; it has become increasingly important as an agent of diarrhea in human beings&#44; recognizing as potential routes of infection the consumption and manipulation of contaminated raw or poorly cooked food of animal origin and untreated water<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">2&#44;5&#44;13</span></a>&#46;</p><p id="par0015" class="elsevierStylePara elsevierViewall">One of the main reservoirs of this species is water in diverse environments<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">2</span></a> where they may interact with other microorganisms&#44; which are natural inhabitants of hydric ecosystems&#44; in particular with free-living amoebas&#46;</p><p id="par0020" class="elsevierStylePara elsevierViewall">Free-living amoebas of the Genus <span class="elsevierStyleItalic">Acanthamoeba</span> are widely distributed in water environments&#46; Amoebic trophozoites are part of the indigenous inhabitants of natural water bodies&#44; sharing this habitat with other microorganisms&#44; mainly bacteria&#46; Since these bacteria can survive for diverse periods within the amoeba&#44; many researchers have suggested that these protozoa may be a reservoir for bacterial species<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">1</span></a>&#46; Because of the particular ability of some bacteria to survive within amoebae&#44; the members of genus <span class="elsevierStyleItalic">Acanthamoeba</span> could play a role in the transmission of some enteric bacteria&#46; The survival of <span class="elsevierStyleItalic">Campylobacter jejuni</span> inside <span class="elsevierStyleItalic">Acanthamoeba</span> and its transmission to chicken as an endosymbiont of <span class="elsevierStyleItalic">A&#46; castellanii</span> and <span class="elsevierStyleItalic">Acanthamoeba polyphaga</span><a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">1</span></a> has been demonstrated&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">Since <span class="elsevierStyleItalic">A&#46; butzleri</span> is a bacterium that can be recovered from water environments and food products&#44; such as mussels produced in estuarine environments<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">2&#44;6</span></a>&#44; it could share this environment with free-living amoebas&#44; probably establishing close relationships with these protozoa&#44; similarly to those described in the genus <span class="elsevierStyleItalic">Campylobacter</span><a class="elsevierStyleCrossRefs" href="#bib0080"><span class="elsevierStyleSup">1&#44;11&#44;12</span></a>&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">In order to demonstrate if <span class="elsevierStyleItalic">A&#46; butzleri</span> is capable of establishing similar close relations with free-living amoebas&#44; we determined the survival capacity of <span class="elsevierStyleItalic">Arcobacter</span> inside <span class="elsevierStyleItalic">A&#46; castellanii&#46;</span></p><p id="par0035" class="elsevierStylePara elsevierViewall">Three strains of <span class="elsevierStyleItalic">A&#46; butzleri</span> were used for this study&#46; Strain F215 &#40;isolated from child human stools&#41;&#44; strain Puar190 &#40;isolated from chicken meat&#41; and the reference strain LMG10828 &#40;isolated from human feces&#41; used as control strain and kindly provided by Dr&#46; Peter Vandamme &#40;Laboratory of Microbiology&#44; Ghent University&#44; Belgium&#41;&#46; These strains were stored at &#8722;80<span class="elsevierStyleHsp" style=""></span>&#176;C in brain-heart infusion broth supplemented with 15<span class="elsevierStyleHsp" style=""></span>&#37; glycerol&#46; Before each experiment&#44; bacteria were grown on blood agar plates incubated at 26<span class="elsevierStyleHsp" style=""></span>&#176;C for 48<span class="elsevierStyleHsp" style=""></span>h under aerobic conditions&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">A&#46; castellanii</span> strain BP91&#47;2760 was originally isolated from the eye of a patient with keratitis and was kindly provided by Dr&#46; Winiecka-Krusnell &#40;Public Health Agency of Sweden&#41;&#46; Amoebae were maintained axenically in peptone yeast extract glucose &#40;PYG&#41; medium at 26<span class="elsevierStyleHsp" style=""></span>&#176;C as monolayers in 25<span class="elsevierStyleHsp" style=""></span>cm<span class="elsevierStyleSup">2</span> flasks and were examined under an inverted microscope before use&#46; To assess the number of amoebae&#44; counts were made using a Neubauer chamber&#46;</p><p id="par0045" class="elsevierStylePara elsevierViewall">To establish <span class="elsevierStyleItalic">A&#46; butzleri</span>&#47;<span class="elsevierStyleItalic">A&#46; castellanii</span> early relationships&#44; <span class="elsevierStyleItalic">A&#46; castellanii</span> were grown to confluence in 25<span class="elsevierStyleHsp" style=""></span>cm<span class="elsevierStyleSup">2</span> flasks with 5<span class="elsevierStyleHsp" style=""></span>ml of PYG medium&#46; The flask were gently shaken and the PYG with non-adherent amoebae was removed and replaced with fresh medium&#59; then the amoebae were incubated on ice for 30<span class="elsevierStyleHsp" style=""></span>min to weaken adherence&#46; The suspensions were centrifuged at 203<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span> for 10<span class="elsevierStyleHsp" style=""></span>min and the pellet was washed three times with Page&#39;s saline solution&#46; After the last centrifugation&#44; the pellet was suspended in PYG and the total number of amoebae was estimated&#46; Aliquots of 1<span class="elsevierStyleHsp" style=""></span>ml with approximately 6<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">4</span> trophozoites were placed in each well of a 24-well tissue culture dish&#46; Bacterial strains suspended in Page&#39;s saline solution to a density of approximately 1&#46;2<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">8</span> colony forming units &#40;CFU&#41;&#44; and 100<span class="elsevierStyleHsp" style=""></span>ml of this suspension was added to each well containing the amoebae and the plate was incubated at 26<span class="elsevierStyleHsp" style=""></span>&#176;C under aerobic conditions to induce the infection of the amoebas by <span class="elsevierStyleItalic">Arcobacter</span> strains in a 1&#58;2000 ratio&#46; As bacterial survival controls&#44; three wells were inoculated with <span class="elsevierStyleItalic">A&#46; butzleri</span> suspended in Page&#39;s saline solution without amoebae&#46; Finally&#44; the assessment of the early interactions between bacteria and amoebae was performed by taking aliquots of the suspension at 30 and 50<span class="elsevierStyleHsp" style=""></span>min and observed directly under phase contrast microscope&#46;</p><p id="par0050" class="elsevierStylePara elsevierViewall">Long-term interaction between bacteria and amoebae was studied at 24&#44; 48&#44; 72&#44; 120&#44; 168&#44; and 240<span class="elsevierStyleHsp" style=""></span>h using a modification of the gentamicin protection assay proposed by Dirks and Quinlan<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">3</span></a>&#46; In brief&#44; after the early interaction assay&#44; 100<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml gentamicin were added to each well and incubated at room temperature for 1<span class="elsevierStyleHsp" style=""></span>h to kill extra-amoebic bacterial cells&#46; Amoebas were harvested from the well and washed three times with Page&#39;s saline solution&#46; After the third wash&#44; an aliquot of 100<span class="elsevierStyleHsp" style=""></span>&#956;l was taken and plated on blood agar plates to determine the number of extracellular bacteria that might have escaped the gentamicin treatment at each sampling time&#46; 100<span class="elsevierStyleHsp" style=""></span>&#956;l of 0&#46;5&#37; sodium desoxycholate was added to another aliquot for 25&#8211;30<span class="elsevierStyleHsp" style=""></span>min to perform the amoeba lysis&#46; An aliquot of 100<span class="elsevierStyleHsp" style=""></span>&#956;l of the lysate suspension was taken to count the number of total bacteria by CFU counts &#40;intracellular<span class="elsevierStyleHsp" style=""></span>&#43;<span class="elsevierStyleHsp" style=""></span>extracellular bacteria&#41;&#46; Abundance of viable intracellular bacteria was determined by subtracting the number of extracellular bacteria from the total count&#46; All these experiments were carried out in triplicate&#46; The amoebae&#47;bacteria interaction was also monitored and documented through phase contrast microscopy and by bright-field microscopy&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">The results of the assays were divided into two groups in accordance with the duration of the tests performed&#46;</p><p id="par0060" class="elsevierStylePara elsevierViewall">A&#46; &#8211; Early interactions between <span class="elsevierStyleItalic">A&#46; butzleri</span> and <span class="elsevierStyleItalic">A&#46; castellanii</span>&#46; In the first 30<span class="elsevierStyleHsp" style=""></span>min of co-incubation&#44; <span class="elsevierStyleItalic">A&#46; butzleri</span> could be seen in close association with <span class="elsevierStyleItalic">A&#46; castellanii</span>&#44; at this time most of the bacteria showed a tendency to gather at one pole of the amoeba cell &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>A&#41;&#46; Soon after 50<span class="elsevierStyleHsp" style=""></span>min of co-culture&#44; <span class="elsevierStyleItalic">A&#46; butzleri</span> cells were detected inside the amoeba&#46; The internalized <span class="elsevierStyleItalic">A&#46; butzleri</span> were located in vacuoles and never in the amoeba cytoplasm &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>B&#41;&#46; These observations are consistent with our previous studies<a class="elsevierStyleCrossRefs" href="#bib0110"><span class="elsevierStyleSup">7&#44;8</span></a>&#44; confirming an initial contact pattern in the interaction <span class="elsevierStyleItalic">A&#46; butzleri</span>&#47;<span class="elsevierStyleItalic">A&#46; castellanii&#46;</span></p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0065" class="elsevierStylePara elsevierViewall">The internalization of <span class="elsevierStyleItalic">A&#46; butzleri</span> seems to be an active and probable metabolic process since all intracellular bacteria are located in vacuoles and never inside the amoeba cytoplasmic region&#46; These images are similar to those previously reported by our group<a class="elsevierStyleCrossRefs" href="#bib0110"><span class="elsevierStyleSup">7&#44;8</span></a>&#46;</p><p id="par0070" class="elsevierStylePara elsevierViewall">This early observation indicated that the interaction between bacteria and amoebae could be a common phenomenon in <span class="elsevierStyleItalic">Arcobacter</span>&#44; which is time-efficient&#46; Recently&#44; Medina <span class="elsevierStyleItalic">et al&#46;</span><a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">8</span></a> have reported that the attachment process of <span class="elsevierStyleItalic">A&#46; butzleri</span> to <span class="elsevierStyleItalic">A&#46; castellanii</span> involves the participation of mannose-binding proteins and membrane-associated receptors of glucose and galactose present in the amoebae&#44; whereas in their internalization&#44; the protozoan actin polymerization plays an active role&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">B&#46; &#8211; Time course of the interaction of <span class="elsevierStyleItalic">A&#46; butzleri</span> and <span class="elsevierStyleItalic">A&#46; castellanii</span>&#46; The long-term experiments demonstrated that <span class="elsevierStyleItalic">A&#46; butzleri</span> strains F-215 and PUAr190 were able to survive for at least 240<span class="elsevierStyleHsp" style=""></span>h inside the amoebas&#46; On the other hand&#44; control assays done to establish whether the presence of <span class="elsevierStyleItalic">A&#46; butzleri</span> could affect the survival of the amoebae&#44; showed no significant differences in the survival rates of the amoebae with and without bacteria &#40;data not shown&#41;&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">Regarding the survival of <span class="elsevierStyleItalic">A&#46; butzleri</span> within <span class="elsevierStyleItalic">A&#46; castellanii</span> trophozoites at different time points&#44; all the bacteria studied in the co-culture with the amoeba as well as control bacteria showed a decrease in CFU values until reaching the minimum count on the tenth day&#46; There is a clear difference in the survival of these three strains of <span class="elsevierStyleItalic">A&#46; butzleri</span> inside the amoebae&#46; The data suggest that recently isolated <span class="elsevierStyleItalic">A&#46; butzleri</span> strains F-215 and PUAr190 have better survival than the reference strain LMG10828&#46; Their colony counts suggest that this strain does not multiply inside <span class="elsevierStyleItalic">A&#46; castellani</span>&#44; not reaching the high numbers seen for the other two strains &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>&#41;&#46; Furthermore&#44; their UFC numbers drop in a similar way as the number of viable bacterial cells decays in the control assay without amoebae&#46; Whether those differences are related to the source of isolation&#44; geographic origin or number of subcultures is unknown&#44; requiring further investigations&#46;</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia><p id="par0085" class="elsevierStylePara elsevierViewall">The methodology used in this study to estimate the intra-amoeba bacterial survival rate was selected because other methods&#44; such as vital stains and counting by microscopy&#44; express their results as percentages and do not allow to determine the bacterial number as accurately as the gentamicin protection assay does<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">3</span></a>&#46;</p><p id="par0090" class="elsevierStylePara elsevierViewall">The <span class="elsevierStyleItalic">A&#46; butzleri</span> strains used in this study were not only able to infect <span class="elsevierStyleItalic">A&#46; castellanii</span> but also capable of surviving inside the vacuoles&#44; in which we confirmed that bacteria were actively motile&#46; Through the phagolysosome formation analysis&#44; Medina <span class="elsevierStyleItalic">et al&#46;</span><a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">8</span></a> concluded that the survival of <span class="elsevierStyleItalic">A&#46; butzleri</span> inside the amoebae could be related to their ability to remain inside vacuoles not fused with lysosomes&#44; or with their ability to retard phagolysosome formation&#46; However&#44; further investigations are needed to elucidate the survival mechanisms of <span class="elsevierStyleItalic">A&#46; butzleri</span> inside free-living amoebae&#46;</p><p id="par0095" class="elsevierStylePara elsevierViewall">Transmissibility of enteropathogens&#44; particularly those associated in zoonosis&#44; has been difficult to identify and for the same reason&#44; nearly impossible to prevent&#46; In the last quarter of the 20<span class="elsevierStyleSup">th</span> century and the beginning of this century&#44; emerging pathogens have become a major health problem<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">4</span></a>&#46; Recent observations showed that free-living amoebae may act as a reservoir and vehicle for the survival and transmission of <span class="elsevierStyleItalic">C&#46; jejuni</span> and other enteropathogens&#44; providing a unique form of self-perpetuation of these pathogens<a class="elsevierStyleCrossRefs" href="#bib0080"><span class="elsevierStyleSup">1&#44;9&#44;10&#44;12</span></a>&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">Our team has been very interested in the role of <span class="elsevierStyleItalic">Arcobacter</span> species as human pathogens and its association with animal sources<a class="elsevierStyleCrossRefs" href="#bib0100"><span class="elsevierStyleSup">5&#44;6</span></a>&#46; Different studies have indicated that <span class="elsevierStyleItalic">A&#46; butzleri</span> can survive under several environmental conditions and can be isolated from diverse hosts like farm animals&#44; seafood&#44; and hose pets<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">2&#44;5&#44;6</span></a>&#46; In the present study&#44; the survival of <span class="elsevierStyleItalic">A&#46; butzleri</span> inside <span class="elsevierStyleItalic">A&#46; castellanii</span> strongly suggests that this protozoon can serve as a reservoir for <span class="elsevierStyleItalic">A&#46; butzleri</span> and it might be an important vehicle in the transmission of this human emergent zoonotic pathogen&#44; acting as a Trojan horse<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">2&#44;8&#44;12</span></a>&#46; Experimental transmissibility assays of <span class="elsevierStyleItalic">A&#46; butzleri</span> inside <span class="elsevierStyleItalic">A&#46; castellanii</span> into mice&#44; currently performed in our laboratory &#40;data not shown&#41;&#44; allow to infer the eventual participation of this protozoon as transmission vehicle of <span class="elsevierStyleItalic">A&#46; butzleri</span>&#46; Further studies have to be carried out to elucidate this epidemiological angle of the <span class="elsevierStyleItalic">A&#46; butzleri</span>&#47;free-living amoebae relationships&#46;</p><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Ethical disclosures</span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Protection of human and animal subjects</span><p id="par0105" class="elsevierStylePara elsevierViewall">The authors declare that no experiments were performed on humans or animals for this study&#46;</p></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Confidentiality of data</span><p id="par0110" class="elsevierStylePara elsevierViewall">The authors declare that no patient data appear in this article&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Right to privacy and informed consent</span><p id="par0115" class="elsevierStylePara elsevierViewall">The authors declare that no patient data appear in this article&#46;</p></span></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Financial support</span><p id="par0120" class="elsevierStylePara elsevierViewall">Grants DID-UACH S-2007-37 and FONDECYT 1110202&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Conflict of interests</span><p id="par0125" class="elsevierStylePara elsevierViewall">The authors declare that they have no conflict of interests&#46;</p></span></span>"
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        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">The survival of three <span class="elsevierStyleItalic">Arcobacter butzleri</span> strains inside <span class="elsevierStyleItalic">Acanthamoeba castellanii</span> was assessed using axenic cultures of <span class="elsevierStyleItalic">A&#46; castellanii</span> that were inoculated with the tested strains and incubated at 26<span class="elsevierStyleHsp" style=""></span>&#176;C under aerobic conditions for 240<span class="elsevierStyleHsp" style=""></span>h&#46; The behavior of bacteria in contact with amoebae was monitored using phase contrast microscopy&#46; The bacterial survival rate within amoebae was assessed through counting colony forming units&#44; using the gentamicin protection assay&#46; All <span class="elsevierStyleItalic">A&#46; butzleri</span> strains were able to survive during 240<span class="elsevierStyleHsp" style=""></span>h within the amoebae&#44; thus suggesting that &#40;i&#41; <span class="elsevierStyleItalic">A&#46; butzleri</span> resists the amoebic digestion processes at least for the analyzed time&#59; &#40;ii&#41; that <span class="elsevierStyleItalic">A&#46; castellanii</span> could serve as an environmental reservoir for this bacterium&#44; probably acting as a transmission vehicle for <span class="elsevierStyleItalic">A&#46; butzleri</span>&#46;</p></span>"
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        "resumen" => "<span id="abst0010" class="elsevierStyleSection elsevierViewall"><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Se determin&#243; la sobrevida de 3 cepas de <span class="elsevierStyleItalic">Arcobacter butzleri</span> en el interior de trofozo&#237;tos de <span class="elsevierStyleItalic">Acanthamoeba castellanii</span> utilizando cultivos ax&#233;nicos de la ameba&#44; los que fueron inoculados con las cepas bacterianas e incubados a 26<span class="elsevierStyleHsp" style=""></span>&#176;C en aerobiosis durante 240<span class="elsevierStyleHsp" style=""></span>h&#46; La interacci&#243;n bacterias&#47;amebas fue monitorizada mediante microscop&#237;a de contraste de fase&#46; Las tasas de sobrevida bacteriana en el interior de las amebas fueron establecidas a trav&#233;s del recuento de unidades formadoras de colonias mediante ensayos de protecci&#243;n con gentamicina&#46; Todas las cepas de <span class="elsevierStyleItalic">A&#46; butzleri</span> fueron capaces de sobrevivir en el interior de la ameba al menos durante 240<span class="elsevierStyleHsp" style=""></span>h&#44; lo que sugiere&#44; por un lado&#44; que <span class="elsevierStyleItalic">A&#46; butzleri</span> puede resistir los procesos de digesti&#243;n propios de la ameba&#44; al menos por el lapso aqu&#237; analizado y&#44; por el otro&#44; que <span class="elsevierStyleItalic">A&#46; castellanii</span> podr&#237;a actuar como reservorio ambiental de esta bacteria y&#44; probablemente&#44; como veh&#237;culo de transmisi&#243;n de <span class="elsevierStyleItalic">A&#46; butzleri</span>&#46;</p></span>"
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          "en" => "<p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Phase contrast microscopy&#46; &#40;A&#41; Trophozoite of <span class="elsevierStyleItalic">A&#46; castellanii</span> showing <span class="elsevierStyleItalic">A&#46; butzleri</span> seen in close association with <span class="elsevierStyleItalic">A&#46; castellanii</span> gathering at one pole of the amoeba cell known as food-cup &#40;30<span class="elsevierStyleHsp" style=""></span>min of co-incubation&#41;&#46; &#40;B&#41; Trophozoite of <span class="elsevierStyleItalic">A&#46; castellanii</span> showing <span class="elsevierStyleItalic">A&#46; butzleri</span> within the amoebic vacuoles &#40;50<span class="elsevierStyleHsp" style=""></span>min of co-incubation&#41;&#46;</p>"
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Article information

ISSN: 03257541
Original language: English

DOI:10.1016/j.ram.2015.12.003

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Year/Month	Html	Pdf	Total

2024 November	1	0	1
2024 October	20	0	20
2024 September	28	1	29
2024 August	24	2	26
2024 July	16	4	20
2024 June	13	0	13
2024 May	20	1	21
2024 April	24	4	28
2024 March	34	5	39
2024 February	30	6	36
2024 January	41	8	49
2023 December	44	7	51
2023 November	53	6	59
2023 October	67	7	74
2023 September	42	0	42
2023 August	51	4	55
2023 July	53	7	60
2023 June	48	7	55
2023 May	85	28	113
2023 April	78	7	85
2023 March	70	3	73
2023 February	53	4	57
2023 January	57	2	59
2022 December	53	6	59
2022 November	53	11	64
2022 October	43	14	57
2022 September	57	17	74
2022 August	42	9	51
2022 July	49	7	56
2022 June	38	5	43
2022 May	24	12	36
2022 April	20	8	28
2022 March	42	10	52
2022 February	22	4	26
2022 January	32	6	38
2021 December	37	11	48
2021 November	40	5	45
2021 October	51	15	66
2021 September	46	8	54
2021 August	39	6	45
2021 July	24	14	38
2021 June	36	7	43
2021 May	44	11	55
2021 April	142	29	171
2021 March	52	15	67
2021 February	50	13	63
2021 January	37	14	51
2020 December	47	10	57
2020 November	37	9	46
2020 October	25	5	30
2020 September	17	10	27
2020 August	28	11	39
2020 July	14	5	19
2020 June	19	9	28
2020 May	33	7	40
2020 April	17	7	24
2020 March	18	10	28
2020 February	24	10	34
2020 January	22	4	26
2019 December	19	10	29
2019 November	27	5	32
2019 October	24	5	29
2019 September	32	5	37
2019 August	16	5	21
2019 July	32	10	42
2019 June	69	23	92
2019 May	134	23	157
2019 April	62	9	71
2019 March	18	5	23
2019 February	19	3	22
2019 January	11	2	13
2018 December	12	4	16
2018 November	40	9	49
2018 October	30	7	37
2018 September	28	10	38
2018 August	6	2	8
2018 July	9	2	11
2018 June	17	4	21
2018 May	7	7	14
2018 April	16	1	17
2018 March	12	1	13
2018 February	10	3	13
2018 January	16	2	18
2017 December	10	0	10
2017 November	15	4	19
2017 October	9	1	10
2017 September	13	1	14
2017 August	10	4	14
2017 July	9	3	12
2017 June	14	2	16
2017 May	20	1	21
2017 April	12	13	25
2017 March	17	30	47
2017 February	27	1	28
2017 January	14	0	14
2016 December	25	4	29
2016 November	29	8	37
2016 October	28	12	40
2016 September	47	10	57
2016 August	69	6	75
2016 July	44	10	54

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