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Rosa Bragulat, Gemma Castellá, Marcos Isidoro-Ayza, Mariano Domingo, F. Javier Cabañes" "autores" => array:5 [ 0 => array:3 [ "nombre" => "M. Rosa" "apellidos" => "Bragulat" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] 1 => array:3 [ "nombre" => "Gemma" "apellidos" => "Castellá" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] 2 => array:3 [ "nombre" => "Marcos" "apellidos" => "Isidoro-Ayza" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">b</span>" "identificador" => "aff0010" ] ] ] 3 => array:3 [ "nombre" => "Mariano" "apellidos" => "Domingo" "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">b</span>" "identificador" => "aff0010" ] 1 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">c</span>" "identificador" => "aff0015" ] ] ] 4 => array:4 [ "nombre" => "F. Javier" "apellidos" => "Cabañes" "email" => array:1 [ 0 => "javier.cabanes@uab.es" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">*</span>" "identificador" => "cor0005" ] ] ] ] "afiliaciones" => array:3 [ 0 => array:3 [ "entidad" => "Veterinary Mycology Group, Departament of Animal Health and Anatomy, Veterinary School, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Servei de Diagnòstic de Patologia Veterinària, Departament of Animal Health and Anatomy, Veterinary School, Universitat Autònoma de Barcelona, Barcelona, Spain" "etiqueta" => "b" "identificador" => "aff0010" ] 2 => array:3 [ "entidad" => "Centre de Recerca en Sanitat Animal (CReSA), IRTA, Bellaterra, Barcelona, Spain" "etiqueta" => "c" "identificador" => "aff0015" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Tipificación y análisis filogenético de una cepa de <span class="elsevierStyleItalic">Cunninghamella bertholletiae</span> aislada de un delfín mular <span class="elsevierStyleItalic">(Tursiops truncatus)</span>" ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0020" "etiqueta" => "Fig. 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 3107 "Ancho" => 3502 "Tamanyo" => 621244 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0070" class="elsevierStyleSimplePara elsevierViewall">Molecular phylogenetic tree inferred from maximum likelihood analysis of ITS sequences. Bootstrap replications frequencies over 70% (1000 replications) are shown at nodes. The tree is rooted with <span class="elsevierStyleItalic">Abisidia cuneospora</span>.</p>" ] ] ] "textoCompleto" => "<span class="elsevierStyleSections"><p id="par0005" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">Cunninghamella</span> is a genus of the order Mucorales which includes saprophytic species, rarely causing mycoses. The more common pathogens of this order are included in the genera <span class="elsevierStyleItalic">Rhizopus</span>, <span class="elsevierStyleItalic">Mucor</span> and <span class="elsevierStyleItalic">Lichtheimia</span>.<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">5</span></a> Currently 12 species are accepted in <span class="elsevierStyleItalic">Cunninghamella</span>,<a class="elsevierStyleCrossRef" href="#bib0130"><span class="elsevierStyleSup">13</span></a> of which the most frequently reported in human mycoses is the thermophilic species <span class="elsevierStyleItalic">Cunninghamella bertholletiae</span>. This species remains a rare cause of human mucormycosis and has been described almost exclusively for immunosuppressed hosts.<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">5</span></a> Infections by other species such <span class="elsevierStyleItalic">Cunninghamella blakesleeana</span>, <span class="elsevierStyleItalic">Cunninghamella echinulata</span>, and <span class="elsevierStyleItalic">Cunninghamella elegans</span> are highly exceptional.<a class="elsevierStyleCrossRef" href="#bib0125"><span class="elsevierStyleSup">12</span></a> In the same way this species is not often reported as an animal pathogen. A comprehensive review of opportunistic mycoses of animals (humans included) was published by Smith<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">9</span></a> in the late eighties of the last century. This book reviews important aspects of the pathogenesis of these mycoses and includes a detailed list of probable cases of mucormycosis in more than 40 animal species other than man. Although this fungus is considered in this book as a well documented causal agent of mucormycosis in the human being, none of the cases mentioned in animals was caused by <span class="elsevierStyleItalic">C. bertholletiae</span>. So this species does not appear to cause mucormycosis in animals.</p><p id="par0010" class="elsevierStylePara elsevierViewall">Surprisingly, more recently this fungus has been involved in two cases of mucormycosis in cetaceans. <span class="elsevierStyleItalic">C. bertholletiae</span> has been reported as the cause of fatal pneumonia in a captive-held killer whale (<span class="elsevierStyleItalic">Orcinus orca</span>)<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">1</span></a> and as the cause of meningoencephalomyelitis in a freeranging bottlenose dolphin (<span class="elsevierStyleItalic">Tursiops truncatus</span>), which was found stranded and dead on the Spanish Mediterranean coast.<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">6</span></a> Until now, to our knowledge, no other cases of <span class="elsevierStyleItalic">C. bertholletiae</span> infection, neither in domestic nor in wild animals, have been published.</p><p id="par0015" class="elsevierStylePara elsevierViewall">In this paper we describe the phenotypic and genotypic characterization, and the phylogenetic analysis of the isolate of <span class="elsevierStyleItalic">C. bertholletiae</span> involved in the first case of a central nervous system mucormycosis in a bottlenose dolphin.</p><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Materials and methods</span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Morphological and physiological characterization</span><p id="par0020" class="elsevierStylePara elsevierViewall">The morphological and physiological characterization of the isolate was carried out based on macroscopic and microscopic characteristics of the colonies on Sabouraud glucose agar (SGA; Oxoid, Basingstoke, UK) and potato dextrose agar (PDA; Becton Dickinson) cultures after incubation at 25<span class="elsevierStyleHsp" style=""></span>°C for 7 days. Other tests, such as the isolate temperature tolerance (30, 40 and 45<span class="elsevierStyleHsp" style=""></span>°C) on SGA and PDA for 7 days, were also performed.<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">4,13</span></a></p></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">DNA extraction, gene amplification, sequencing and phylogenetic analysis</span><p id="par0025" class="elsevierStylePara elsevierViewall">Molecular characterization of the isolate was carried out based on the D1/D2 regions of the 26S (D1/D2) and the ITS-5.8S (ITS) rRNA gene sequencing. DNA was extracted and purified directly from a seven day old culture in SGA according to the FastDNA Spin kit protocol with the FastPrep FP-24 instrument. The procedures for the amplification and sequencing were according to the protocols described previously.<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">2</span></a> The resulting sequences were aligned using Clustal X v2.0.12<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">7</span></a> and regions of ambiguous alignment were removed with Gblocks.<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">3</span></a> Maximum likelihood trees were inferred with the server version of RAxML-HPC2 v8,<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">10</span></a> as implemented on the Cipres portal, using the GTR model. The robustness of the trees was estimated by a bootstrap analysis with 1000 replicates. For the phylogenetic analyses sequences of representative strains from the rest of the <span class="elsevierStyleItalic">Cunninghamella</span> species deposited in GenBank were also included.</p></span></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Results</span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Morphology and physiology</span><p id="par0030" class="elsevierStylePara elsevierViewall">Colonies on SGA and PDA were fast-growing, white at first, although they became tannish-gray, covering the whole plate (85<span class="elsevierStyleHsp" style=""></span>mm in diameter) after 7 days of incubation at 25, 30 and 40<span class="elsevierStyleHsp" style=""></span>°C. Limited growth was observed after 7 days at 45<span class="elsevierStyleHsp" style=""></span>°C (20<span class="elsevierStyleHsp" style=""></span>mm in diameter) (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>). The micromorphology showed characteristic erect sporangiophores showing the typical branching patterns. The sporangiophores had short lateral branches in the apical region which ended in globose to pyriform vesicles covered with globose one-spored sporangioles. Sporangiola born on pedicels, and vesicles were ovoid to subglobose, 6.4–11.2<span class="elsevierStyleHsp" style=""></span>×<span class="elsevierStyleHsp" style=""></span>5.4–8.2<span class="elsevierStyleHsp" style=""></span>μm, hyaline to pale brownish and smooth to very shortly echinulate (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>). Chlamydospores were not observed. The isolate was identified as <span class="elsevierStyleItalic">C. bertholletiae</span> on the basis of its morphological and thermophilic characteristics.<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">4,13</span></a></p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><elsevierMultimedia ident="fig0010"></elsevierMultimedia></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Molecular and phylogenetic analysis</span><p id="par0035" class="elsevierStylePara elsevierViewall">The D1/D2 and ITS sequences included 693 and 688 base pairs, respectively. The nucleotide sequences of the D1/D2 and ITS determined in this study were deposited in the GenBank and their accession numbers are given in <a class="elsevierStyleCrossRefs" href="#fig0015">Figs. 3 and 4</a>. In the phylogenetic trees of D1/D2 (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>) and ITS (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>), our isolate clustered with other <span class="elsevierStyleItalic">C. bertholletiae</span> strains forming a well supported clade (100% bootstrap). Our strain and <span class="elsevierStyleItalic">C. bertholletiae</span> CBS 779.68 showed the same D1/D2 sequence and differed at 1–2<span class="elsevierStyleHsp" style=""></span>bp from the rest of the strains analyzed. Intraspecific variability in ITS sequences was greater. Our strain differed from neotype strain NRRL 1380 of <span class="elsevierStyleItalic">C. bertholletiae</span> at 44 nucleotide positions whereas only differed at 3 nucleotide positions from <span class="elsevierStyleItalic">C. bertholletiae</span> ATCC 42115.</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia><elsevierMultimedia ident="fig0020"></elsevierMultimedia></span></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Discussion</span><p id="par0040" class="elsevierStylePara elsevierViewall">The <span class="elsevierStyleItalic">C. bertholletiae</span> isolate studied was unambiguously identified by its morphology and genotypic methods. Morphological and physiological characters matched those described for this species.<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">4,13</span></a><span class="elsevierStyleItalic">C. bertholletiae</span> is quite different morphologically from <span class="elsevierStyleItalic">Cunninghamella intermedia</span> and <span class="elsevierStyleItalic">Cunninghamella multiverticillata</span> which have a similar temperature tolerance. They are able to grow at 41–43<span class="elsevierStyleHsp" style=""></span>°C (maximum growth temperature).<a class="elsevierStyleCrossRef" href="#bib0130"><span class="elsevierStyleSup">13</span></a> Despite the fact that the latter two are thermophilic species, <span class="elsevierStyleItalic">C. intermedia</span> and <span class="elsevierStyleItalic">C. multiverticillata</span> are not involved in human or animal infections. However <span class="elsevierStyleItalic">C. bertholletiae</span> grows at 45<span class="elsevierStyleHsp" style=""></span>°C<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">4</span></a> and is the most frequently reported <span class="elsevierStyleItalic">Cunninghamella</span> species in human mycoses.</p><p id="par0045" class="elsevierStylePara elsevierViewall">The identification of the isolate was confirmed by DNA sequencing of the D1/D2 and ITS. In the phylogenetic study, the isolate clustered in the same clade as <span class="elsevierStyleItalic">C. bertholletiae</span> NRRL 1380 neotype strain although some differences were observed in the ITS. It has been reported that ITS regions are highly variable within the genus <span class="elsevierStyleItalic">Cunninghamella</span>, showing an intraspecific variability that ranges from 2.9% to 14.4%, sometimes even greater than interspecific distances between closely related species.<a class="elsevierStyleCrossRef" href="#bib0125"><span class="elsevierStyleSup">12</span></a> The intraspecific variability of ITS sequences described for <span class="elsevierStyleItalic">C. bertholletiae</span> is 1%,<a class="elsevierStyleCrossRef" href="#bib0120"><span class="elsevierStyleSup">11</span></a> although the neotype strain NRRL 1380 was not included in the study. The strain in our study had an identity of 99% when compared to different strains of <span class="elsevierStyleItalic">C. bertholletiae</span> isolated from different human patients, as well as strain ATCC 42115. This strain showed 100% identity with a clinical strain isolated from a pulmonary case of mucormycosis in a killer whale.<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">1</span></a></p><p id="par0050" class="elsevierStylePara elsevierViewall">Mucormycosis in animals is frequently recorded in association with erosion and/or ulceration of the alimentary tract, placentitis and acute pneumonia, among other sites of infection. In ruminants, increased gastric acidity, malnutrition and the feeding of rough, dry food may lead to the development of gastrointestinal mucormycotic lesions.<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">9</span></a> However, in animals, as in some human cases, diagnosis of mucormycosis is often still made at necropsy with histopathological evidence of tissue invasion by characteristic non-septate broad hyphae, but no microbiological analyses are performed to identify the etiological agent. <span class="elsevierStyleItalic">C. bertholletiae</span> is recognized as a cosmopolitan soil organism which has been also found in a wide variety of nuts, seeds, and plants. The small size of sporangiospores allows them to remain airborne for prolonged periods, which can increase the exposure risk. The predominant mode of acquisition of <span class="elsevierStyleItalic">C. bertholletiae</span> infections is presumed to be via the respiratory tract.<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">5</span></a></p><p id="par0055" class="elsevierStylePara elsevierViewall">In the cetacean cases, the possible sources of infection are unclear. However, the entry via aerosol into the laryngeal tonsillar tissue and hematogenous dissemination to the central nervous system was suspected in the dolphin case, based on the severe involvement of arterioles at the basal arteriolar network of the brain.<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">6</span></a> On the other hand, in the killer whale case a large number of stones were found in the stomach compartments; they might act as a facilitating factor for the onset of the mucormycosis in this immunocompromised animal.<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">1</span></a></p><p id="par0060" class="elsevierStylePara elsevierViewall">The reasons why this pathogen has been isolated only in cetaceans and not in other domestic or wild animals are at the moment unknown and need further study. However, <span class="elsevierStyleItalic">C. bertholletiae</span> has been recently recovered from gastric fluid and blowhole bottlenose dolphin samples.<a class="elsevierStyleCrossRef" href="#bib0105"><span class="elsevierStyleSup">8</span></a> Surprisingly, it was the only mucoralean fungus isolated from these animals.</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Conflict of interest</span><p id="par0065" class="elsevierStylePara elsevierViewall">The authors declare that they have no conflicts of interest</p></span></span>" "textoCompletoSecciones" => array:1 [ "secciones" => array:10 [ 0 => array:3 [ "identificador" => "xres938733" "titulo" => "Abstract" "secciones" => array:5 [ 0 => array:2 [ "identificador" => "abst0005" "titulo" => "Background" ] 1 => array:2 [ "identificador" => "abst0010" "titulo" => "Aims" ] 2 => array:2 [ "identificador" => "abst0015" "titulo" => "Methods" ] 3 => array:2 [ "identificador" => "abst0020" "titulo" => "Results" ] 4 => array:2 [ "identificador" => "abst0025" "titulo" => "Conclusions" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec912433" "titulo" => "Keywords" ] 2 => array:3 [ "identificador" => "xres938734" "titulo" => "Resumen" "secciones" => array:5 [ 0 => array:2 [ "identificador" => "abst0030" "titulo" => "Antecedentes" ] 1 => array:2 [ "identificador" => "abst0035" "titulo" => "Objetivos" ] 2 => array:2 [ "identificador" => "abst0040" "titulo" => "Métodos" ] 3 => array:2 [ "identificador" => "abst0045" "titulo" => "Resultados" ] 4 => array:2 [ "identificador" => "abst0050" "titulo" => "Conclusiones" ] ] ] 3 => array:2 [ "identificador" => "xpalclavsec912432" "titulo" => "Palabras clave" ] 4 => array:3 [ "identificador" => "sec0005" "titulo" => "Materials and methods" "secciones" => array:2 [ 0 => array:2 [ "identificador" => "sec0010" "titulo" => "Morphological and physiological characterization" ] 1 => array:2 [ "identificador" => "sec0015" "titulo" => "DNA extraction, gene amplification, sequencing and phylogenetic analysis" ] ] ] 5 => array:3 [ "identificador" => "sec0020" "titulo" => "Results" "secciones" => array:2 [ 0 => array:2 [ "identificador" => "sec0025" "titulo" => "Morphology and physiology" ] 1 => array:2 [ "identificador" => "sec0030" "titulo" => "Molecular and phylogenetic analysis" ] ] ] 6 => array:2 [ "identificador" => "sec0035" "titulo" => "Discussion" ] 7 => array:2 [ "identificador" => "sec0040" "titulo" => "Conflict of interest" ] 8 => array:2 [ "identificador" => "xack317087" "titulo" => "Acknowledgments" ] 9 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2016-11-28" "fechaAceptado" => "2017-03-10" "PalabrasClave" => array:2 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec912433" "palabras" => array:5 [ 0 => "Animal" 1 => "Cetacean" 2 => "<span class="elsevierStyleItalic">Cunninghamella bertholletiae</span>" 3 => "Dolphin" 4 => "Mucormycosis" ] ] ] "es" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Palabras clave" "identificador" => "xpalclavsec912432" "palabras" => array:5 [ 0 => "Animales" 1 => "Cetáceos" 2 => "<span class="elsevierStyleItalic">Cunninghamella bertholletiae</span>" 3 => "Delfín" 4 => "Mucormicosis" ] ] ] ] "tieneResumen" => true "resumen" => array:2 [ "en" => array:3 [ "titulo" => "Abstract" "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleItalic">Cunninghamella</span> is a genus of the order Mucorales which includes saprophytic species, rarely causing mycoses. The most frequently reported in human mycoses is the thermophilic species <span class="elsevierStyleItalic">Cunninghamella bertholletiae</span>. However, this species does not appear to cause mucormycosis in animals, so there is scarce information about <span class="elsevierStyleItalic">C. bertholletiae</span> isolates from animals.</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Aims</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">In this paper we describe the phenotypic and genotypic characterization, and the phylogenetic analysis, of an isolate of <span class="elsevierStyleItalic">C. bertholletiae</span> involved in a central nervous system mucormycosis in a dolphin.</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Methods</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">The isolate studied in this publication was characterized using the current morphological and physiological identification system for <span class="elsevierStyleItalic">Cunninghamella</span> species. DNA sequencing and analysis of the D1/D2 regions of the 26S rRNA gene and the ITS-5.8S rRNA gene sequences were also performed.</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Results</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">Colonies were fast-growing, white at first, although they became tannish-gray, covering the whole plate after 7 days of incubation at 30 and 40<span class="elsevierStyleHsp" style=""></span>°C. Limited growth was observed after 7 days at 45<span class="elsevierStyleHsp" style=""></span>°C. The micromorphology showed characteristic erect sporangiophores. The identification of the isolate was confirmed by DNA sequencing of the D1/D2 regions of the 26S and the ITS-5.8S (ITS) rRNA gene sequencing.</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Conclusions</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">In the phylogenetic study, the isolate clustered in the same clade as <span class="elsevierStyleItalic">C. bertholletiae</span> neotype strain although some differences were observed in the ITS sequences. In the cetacean cases, the possible sources of infection are unclear. The reasons why this pathogen has been found only in cetaceans and not in other domestic or wild animals are at the moment unknown and need further study.</p></span>" "secciones" => array:5 [ 0 => array:2 [ "identificador" => "abst0005" "titulo" => "Background" ] 1 => array:2 [ "identificador" => "abst0010" "titulo" => "Aims" ] 2 => array:2 [ "identificador" => "abst0015" "titulo" => "Methods" ] 3 => array:2 [ "identificador" => "abst0020" "titulo" => "Results" ] 4 => array:2 [ "identificador" => "abst0025" "titulo" => "Conclusions" ] ] ] "es" => array:3 [ "titulo" => "Resumen" "resumen" => "<span id="abst0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Antecedentes</span><p id="spar0030" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleItalic">Cunninghamella</span> es un género perteneciente al orden Mucorales, que incluye especies saprófitas que raramente causan micosis. De este género, <span class="elsevierStyleItalic">Cunninghamella bertholletiae</span> es la especie termófila más frecuentemente citada en micosis humanas. No obstante, no parece que sea una causa habitual de mucormicosis en animales, ya que es escasa la información sobre cepas de esta especie procedentes de estos.</p></span> <span id="abst0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Objetivos</span><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">En esta publicación describimos la tipificación fenotípica, genotípica y el análisis filogenético de una cepa de <span class="elsevierStyleItalic">C. bertholletiae</span> causante de una mucormicosis del sistema nervioso central en un delfín.</p></span> <span id="abst0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Métodos</span><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">La cepa fue tipificada mediante los criterios morfológicos y fisiológicos actualmente utilizados para la identificación de estas especies. También se llevó a cabo la secuenciación y el análisis de los fragmentos génicos D1/D2 26S e ITS-5.8S del ARN ribosómico.</p></span> <span id="abst0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Resultados</span><p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">Las colonias presentaron un crecimiento rápido; eran blanquecinas al principio y se volvieron de color marrón agrisado con el tiempo, y cubrieron totalmente las placas a los 7 días de incubación a las temperaturas de 30 y 40<span class="elsevierStyleHsp" style=""></span>°C. A 45<span class="elsevierStyleHsp" style=""></span>°C, después de 7 días de incubación, el crecimiento fue limitado. Al microscopio se pudieron observar los característicos esporangióforos de esta especie. La identificación de la cepa se confirmó mediante la secuenciación de los fragmentos génicos D1/D2 26S e ITS-5.8S del ARN ribosómico.</p></span> <span id="abst0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Conclusiones</span><p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">En el estudio filogenético, la cepa se agrupó en el mismo clado que la cepa neotipo de <span class="elsevierStyleItalic">C. bertholletiae</span>, aunque se detectaron algunas diferencias en las secuencias correspondientes a los ITS. En los casos causados por esta especie en cetáceos, se desconocen las posibles fuentes de infección. Tampoco se conoce por el momento por qué este patógeno ha sido aislado solo de cetáceos y no de otros animales domésticos o salvajes.</p></span>" "secciones" => array:5 [ 0 => array:2 [ "identificador" => "abst0030" "titulo" => "Antecedentes" ] 1 => array:2 [ "identificador" => "abst0035" "titulo" => "Objetivos" ] 2 => array:2 [ "identificador" => "abst0040" "titulo" => "Métodos" ] 3 => array:2 [ "identificador" => "abst0045" "titulo" => "Resultados" ] 4 => array:2 [ "identificador" => "abst0050" "titulo" => "Conclusiones" ] ] ] ] "multimedia" => array:4 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Fig. 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 788 "Ancho" => 1725 "Tamanyo" => 192885 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0055" class="elsevierStyleSimplePara elsevierViewall">Colonial morphology of the isolate of <span class="elsevierStyleItalic">C. bertholletiae</span> on potato dextrose agar after 7 days at (a) 40<span class="elsevierStyleHsp" style=""></span>°C and (b) 45<span class="elsevierStyleHsp" style=""></span>°C.</p>" ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Fig. 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1260 "Ancho" => 1650 "Tamanyo" => 146459 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0060" class="elsevierStyleSimplePara elsevierViewall">Characteristic immature (a) and mature (b) vesicles of the isolate of <span class="elsevierStyleItalic">C. bertholletiae</span>, covered with globose one-spored sporangioles borne on pedicels. Sporangiospores (c). Lactophenol cotton blue stain.</p>" ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Fig. 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 2002 "Ancho" => 3502 "Tamanyo" => 395412 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0065" class="elsevierStyleSimplePara elsevierViewall">Molecular phylogenetic tree inferred from maximum likelihood analysis of D1/D2 sequences. Bootstrap replications frequencies over 70% (1000 replications) are shown at nodes. The tree is rooted with <span class="elsevierStyleItalic">Gongronella butleri</span>.</p>" ] ] 3 => array:7 [ "identificador" => "fig0020" "etiqueta" => "Fig. 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 3107 "Ancho" => 3502 "Tamanyo" => 621244 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0070" class="elsevierStyleSimplePara elsevierViewall">Molecular phylogenetic tree inferred from maximum likelihood analysis of ITS sequences. Bootstrap replications frequencies over 70% (1000 replications) are shown at nodes. 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Financial support came from Servei Veterinari de Bacteriologia i Micologia and Servei de Diagnòstic de Patologia Veterinària of the UAB.</p>" "vista" => "all" ] ] ] "idiomaDefecto" => "en" "url" => "/11301406/0000003400000004/v1_201711120057/S1130140617300530/v1_201711120057/en/main.assets" "Apartado" => array:4 [ "identificador" => "7997" "tipo" => "SECCION" "es" => array:2 [ "titulo" => "Originales" "idiomaDefecto" => true ] "idiomaDefecto" => "es" ] "PDF" => "https://static.elsevier.es/multimedia/11301406/0000003400000004/v1_201711120057/S1130140617300530/v1_201711120057/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S1130140617300530?idApp=UINPBA00004N" ]
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Original article
Characterization and phylogenetic analysis of a Cunninghamella bertholletiae isolate from a bottlenose dolphin (Tursiops truncatus)
Tipificación y análisis filogenético de una cepa de Cunninghamella bertholletiae aislada de un delfín mular (Tursiops truncatus)
M. Rosa Bragulata, Gemma Castelláa, Marcos Isidoro-Ayzab, Mariano Domingob,c, F. Javier Cabañesa,
Autor para correspondencia
a Veterinary Mycology Group, Departament of Animal Health and Anatomy, Veterinary School, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain
b Servei de Diagnòstic de Patologia Veterinària, Departament of Animal Health and Anatomy, Veterinary School, Universitat Autònoma de Barcelona, Barcelona, Spain
c Centre de Recerca en Sanitat Animal (CReSA), IRTA, Bellaterra, Barcelona, Spain