Epidemiological and molecular characteristics of Chlamydia psittaci from 8 human cases of psittacosis and 4 related birds in Argentina

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Guzzetti, Cecilia M. Vescina, M. Florencia Gil, Blanca M. Gatti" "autores" => array:4 [ 0 => array:2 [ "nombre" => "Luciana B." "apellidos" => "Guzzetti" ] 1 => array:2 [ "nombre" => "Cecilia M." "apellidos" => "Vescina" ] 2 => array:2 [ "nombre" => "M. Florencia" "apellidos" => "Gil" ] 3 => array:2 [ "nombre" => "Blanca M." 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"apellidos" => "Cadario" "email" => array:1 [ 0 => "ecadario@anlis.gov.ar" ] "referencia" => array:3 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "◊" "identificador" => "fn1" ] 2 => array:2 [ "etiqueta" => "*" "identificador" => "cor0005" ] ] ] 1 => array:3 [ "nombre" => "María C." "apellidos" => "Frutos" "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] 1 => array:2 [ "etiqueta" => "◊" "identificador" => "fn1" ] ] ] 2 => array:3 [ "nombre" => "Maite B." "apellidos" => "Arias" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 3 => array:3 [ "nombre" => "Javier A." "apellidos" => "Origlia" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "c" "identificador" => "aff0015" ] ] ] 4 => array:3 [ "nombre" => "Vanina" "apellidos" => "Zelaya" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "d" "identificador" => "aff0020" ] ] ] 5 => array:3 [ "nombre" => "María J." "apellidos" => "Madariaga" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "e" "identificador" => "aff0025" ] ] ] 6 => array:3 [ "nombre" => "Claudia S." "apellidos" => "Lara" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] ] ] 7 => array:3 [ "nombre" => "Viviana" "apellidos" => "Ré" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] 8 => array:3 [ "nombre" => "Cecilia G." "apellidos" => "Cuffini" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] ] "afiliaciones" => array:5 [ 0 => array:3 [ "entidad" => "INEI-ANLIS «Dr. Carlos G. Malbrán», Ciudad Autónoma de Buenos Aires, Argentina" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Instituto de Virología «Dr. J.M. Vanella», Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Córdoba, Argentina" "etiqueta" => "b" "identificador" => "aff0010" ] 2 => array:3 [ "entidad" => "Cátedra de Patología de Aves y Pilíferos, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Argentina" "etiqueta" => "c" "identificador" => "aff0015" ] 3 => array:3 [ "entidad" => "UOCCB-ANLIS «Dr. Carlos G. Malbrán», Ciudad Autónoma de Buenos Aires, Argentina" "etiqueta" => "d" "identificador" => "aff0020" ] 4 => array:3 [ "entidad" => "Instituto de Zoonosis Luis Pasteur, Departamento de Diagnóstico y Producción de Biológicos, Ciudad Autónoma de Buenos Aires, Argentina" "etiqueta" => "e" "identificador" => "aff0025" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Características epidemiológicas y moleculares de Chlamydia psittaci provenientes de 8 casos humanos de psitacosis y de 4 aves relacionadas en la Argentina" ] ] "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" => 1644 "Ancho" => 2633 "Tamanyo" => 317511 ] ] "descripcion" => array:1 [ "en" => "Neighbor-joining dendrogram based on comparison of 290bp fragment of the ompA gene in Chlamydia. Numbers above branches are bootstrap values as a percentage of 1000 pseudo-replicates and only bootstrap values >60% are shown. Chlamydia caviae GPIC was used as an out-group. Scale bar shows the percentage sequence diversity." ] ] ] "textoCompleto" => "Psittacosis is a zoonosis caused by Chlamydia psittaci, an obligate intracellular bacterium belonging to the Chlamydiaceae family and its single genus, Chlamydia<a class="elsevierStyleCrossRef" href="#bib0135">13</a>.Human infections vary from unapparent to severe systemic diseases: influenza – like illnesses, severe atypical pneumonia, endocarditis, myocarditis, meningitis, conjunctivitis, and others. The disease is rarely fatal in properly treated patients. Therefore, awareness of an early diagnosis is important. Pulmonary involvement is common and information of recent exposure to birds is frequently omitted during medical consultation.Confirmation by molecular methods and genotyping of C. psittaci infection using human respiratory samples and animal samples is a matter of both diagnostic and epidemiological relevance<a class="elsevierStyleCrossRef" href="#bib0100">6</a>.C. psittaci is divided into at least 9 genotypes (A–F, E/B, M56 and WC). Sequence analysis of the ompA gene which encodes the major outer membrane protein (MOMP) gene, is one way to identify all known and eventual new genotypes<a class="elsevierStyleCrossRef" href="#bib0085">3</a>.Genotypes A–F and E/B are associated with birds and genotypes WC and M56 have been found in cattle and muskrats, respectively<a class="elsevierStyleCrossRef" href="#bib0085">3</a>. Genotype A is associated with Psittaciformes, B with pigeons, C with ducks and geese<a class="elsevierStyleCrossRef" href="#bib0125">11</a>, D with turkeys, E with pigeons and ratites, F with psittacine birds and turkeys<a class="elsevierStyleCrossRef" href="#bib0095">5</a>, and genotype E/B is mainly associated with ducks, turkeys and pigeons<a class="elsevierStyleCrossRefs" href="#bib0085">3,7</a>.The aim of this study was the molecular characterization of C. psittaci polymerase chain reaction-positive (PCR-positive) samples received in the National Reference Laboratory INEI-ANLIS “Dr. Carlos G. Malbrán”, Buenos Aires, Argentina.Eight hundred and fifty-six samples from patients of different cities of Argentina, in whom psittacosis was suspected based on clinical presentation and/or history of exposure with infected birds, were analyzed between January 2010 and March 2015. Eight hundred and forty-six were respiratory samples (nasal and pharyngeal swabs, nasopharyngeal aspirates, tracheal aspirates and bronchioalveolar lavages) and 10 were ocular swabs. Cloacal and ocular swabs from 10 related birds were collected and stored until processed. Dacron swabs were placed in a 2ml sucrose–phosphate–glutamate or UTM (Copan Italia, Brescia, Italy), and stored at 4°C. Two hundred microliters of this suspension were subjected to DNA extraction using the DNeasy Blood & Tissue Kit (Qiagen, Valencia, CA, and USA) according to the manufacturer's instructions. DNA was eluted into 200μl of Qiagen elution buffer and stored at –20°C until tested.A nested PCR protocol was used to amplify a portion of the 16S rRNA gene as previously described by Messmer et al.<a class="elsevierStyleCrossRef" href="#bib0120">10</a> Briefly, 5μl of DNA extract was used in the first step to amplify a 436bp fragment common to the Chlamydia genus. In a second PCR step, 5μl of the PCR product obtained in the previous PCR reaction was used to amplify a 127bp specific product of C. psittaci.Genotyping of C. psittaci was attempted in all positive samples in the 16S rRNA gene PCR. Amplification by nested PCR followed by sequencing of the variable domains III and IV of the ompA gene of C. psittaci was performed as described by Sachse and Hotzel.<a class="elsevierStyleCrossRef" href="#bib0130">12</a> Briefly, 5μl of DNA extract were used in the first step to amplify a 576bp fragment of the ompA gene common to the Chlamydia genus. In a second PCR step, 5μl of the PCR product obtained in the previous PCR reaction were used to amplify a 389bp specific product of C. psittaci.Nested PCR products were purified by gel electrophoresis using the AccuPrep® PCR Purification Kit (USA Bioneer Inc.) and subjected to a direct nucleotide sequencing reaction in both directions using the internal (second-round) PCR primers by Macrogen, Inc. (Seoul, Korea). The sequences obtained from fragments of the ompA gene were edited and prepared with BioEdit V 7.0.9<a class="elsevierStyleCrossRef" href="#bib0090">4</a> and subsequently aligned with Clustal X 2.12<a class="elsevierStyleCrossRef" href="#bib0110">8</a>, along with the sequences downloaded from GenBank. Relatedness of newly characterized sequences was assessed by analysis using the 2.2.19 Basic Local Alignment Search Tool. The choice of the genomic region was based on the fact that it is widely associated with the genetic divergence of C. psittaci and it identifies all known genotypes<a class="elsevierStyleCrossRef" href="#bib0085">3</a>.The dendrogram was constructed using the Tree Explorer module of the MEGA software version 4<a class="elsevierStyleCrossRef" href="#bib0140">14</a> with the neighbor-joining method and the p-distance parameter. The branch support was evaluated by nonparametric bootstrapping with 1000 pseudo-replicas.Among the respiratory and ocular swabs samples of the 856 patients studied, 48 (5.6%) tested positive for C. psittaci by nested-PCR. Additionally, 6 samples from related birds tested positive.Among these 54 positive samples for C. psittaci, only 12 produced enough PCR products in the ompA nested PCR to allow sequencing and genotype determination<a class="elsevierStyleCrossRef" href="#bib0135">13</a> (8 from humans and 4 from birds). The mean age of the infected patients was 33.5 years (r=5–62 years); 62.5% (5/8) were young workers aged 23–45 years. Male/female ratio was 1:1.The analysis of the medical records showed that community-acquired pneumonia (CAP) was the most frequent clinical presentation (50%; 4/8), followed by acute respiratory tract infection (25%; 2/8). Whooping cough was suspected in one case (12.5%; 1/8). One patient (12.5%; 1/8) with conjunctivitis tested positive for C. psittaci.Five patients reported contact with birds, especially parrots (n=4). Contact with a domestic or companion bird was not reported in the remaining three C. psittaci positive patients.Clinical presentation and sources of the eight C. psittaci-positive samples are shown in <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>.<elsevierMultimedia ident="tbl0005"></elsevierMultimedia>Genetic diversity and associations among the detected positive samples of C. psittaci were determined by the sequencing and genetic analysis of the composition of the ompA gene. The sequences obtained in this study were deposited in GenBank under accession numbers KU357040–KU357046; KU357049; KU365350–KU365353.In the present study all C. psittaci-positive samples (from humans and birds) were recognized as belonging to genotype A by phylogenetic analysis (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>). However, in Córdoba, a central region of Argentina, genotype A was detected in only one case (12.5%, 1/8), being WC (75%, 6/8) the genotype most frequently found in patients with suspected human psittacosis<a class="elsevierStyleCrossRef" href="#bib0075">1</a> and in captive birds<a class="elsevierStyleCrossRef" href="#bib0080">2</a>. Probably the differences could be due to different clinical conditions of the patients in both studies or/and to geographical sources. This differential genotypic profile is an interesting finding that should be investigated in the future. C. psittaci genotype A is more often found among psittacine birds such as parrots and cockatiels. However, the most prevalent C. psittaci genotype in human infections is currently unknown<a class="elsevierStyleCrossRef" href="#bib0100">6</a>.<elsevierMultimedia ident="fig0005"></elsevierMultimedia>The analysis of the medical records showed that CAP was the most frequent clinical presentation. In most studies on CAP, psittacosis was diagnosed only with serological tests. PCR leads to a quick diagnosis and lacks broad genus cross-reactivity. Furthermore, PCR products can be used to genotype C. psittaci, which is relevant for public health notification, source detection and control.The use of nucleic acid amplification directly from clinical samples and direct genotyping with phylogenetic analysis in the ompA gene could improve the surveillance and molecular epidemiology of this disease. In this sense, the classical method applying two PCR techniques plus sequencing was successfully used and it should be implemented in regional laboratories for an improved diagnosis of this infectious disease.In this study, a nested PCR and direct sequencing were used to detect and genetically characterize C. psittaci. For the phylogenetic analysis of C. psittaci, we used partial ompA gene amplification. Our results only report the circulation of genotype A associated with respiratory symptoms and conjunctivitis in different areas of our country, at least in our experience with these eight human cases. Psittaciformes especially parakeets and parrots, and individuals in contact with them or with racing pigeons, were the most frequently infected.C. psittaci is rarely suspected in ocular infections, but using molecular techniques, as we did, may be more commonly found<a class="elsevierStyleCrossRef" href="#bib0115">9</a>. In the present study we showed a case of conjunctivitis due to C. psittaci in a 26 year-old woman not responding to the current treatment and living in contact with wild birds.Several limitations of our study need to be considered. Isolation in cell cultures was the preferred technique to confirm and corroborate the results obtained; however, in the case of C. psittaci, this procedure is unadvisable because of the biological risks. Moreover, the phylogenetic analysis showed two sequences corresponding to birds (KU365353 and KU365349) in separated clusters. As can be seen in <a class="elsevierStyleCrossRef" href="#fig0005">Figure 1</a> they have a low degree of homology with others obtained in the same city (74% similarity, one with an avian strain and the other with a human strain) (data not shown). Further studies are needed to confirm the genotype and even the species.Effective public health surveillance for new variants and modes of transmission will be helpful to understanding the evolution and epidemiology of these bacteria. Therefore, new information about sequence variation of subtypes will be needed.Ethical responsibilitiesProtection of human and animal subjectsThe authors state that for this investigation no experiments have been performed on humans or animals.Confidentiality of dataThe authors state that they have followed the protocols of their work center on the publication of patient data.Right to privacy and informed consentThe authors state that in this article there are no patient data.Conflicts of interestThe authors declare that they have no conflicts of interest." "textoCompletoSecciones" => array:1 [ "secciones" => array:8 [ 0 => array:3 [ "identificador" => "xres941457" "titulo" => "Abstract" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0005" ] ] ] 1 => array:2 [ "identificador" => "xpalclavsec914612" "titulo" => "Keywords" ] 2 => array:3 [ "identificador" => "xres941458" "titulo" => "Resumen" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0010" ] ] ] 3 => array:2 [ "identificador" => "xpalclavsec914613" "titulo" => "Palabras clave" ] 4 => array:3 [ "identificador" => "sec0005" "titulo" => "Ethical responsibilities" "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" => "Conflicts of interest" ] 6 => array:2 [ "identificador" => "xack318048" "titulo" => "Acknowledgments" ] 7 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2016-08-01" "fechaAceptado" => "2017-04-10" "PalabrasClave" => array:2 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec914612" "palabras" => array:3 [ 0 => "Chlamydia psittaci" 1 => "Genotype A" 2 => "Psittacosis" ] ] ] "es" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Palabras clave" "identificador" => "xpalclavsec914613" "palabras" => array:3 [ 0 => "Chlamydia psittaci" 1 => "Genotipo A" 2 => "Psitacosis" ] ] ] ] "tieneResumen" => true "resumen" => array:2 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "In Argentina, the epidemiological and molecular characteristics of Chlamydia psittaci infections are still not sufficiently known. A total of 846 respiratory and 10 ocular samples from patients with suspected human psittacosis were tested for C. psittaci from January 2010 to March 2015. Four samples of birds related to these patients were also studied. Forty-eight samples were positive for C. psittaci by a nested PCR. The molecular characterization of twelve C. psittaci PCR-positive samples received in the National Reference Laboratory INEI-ANLIS “Dr. Carlos G. Malbrán”, Buenos Aires, Argentina was performed. Eight positive samples from humans and four from birds were genotyped by ompA gene sequencing. C. psittaci genotype A was found in all human samples and in the related birds. This report contributes to our increasing knowledge of the epidemiological and molecular characteristics of C. psittaci to conduct effective surveillance of its zoonotic infections." ] "es" => array:2 [ "titulo" => "Resumen" "resumen" => "En la Argentina, aún no se conocen suficientemente las características epidemiológicas y moleculares de las infecciones por Chlamydia psittaci. Entre enero del 2010 y marzo del 2015 se estudiaron 846 muestras respiratorias y 10 oculares de pacientes con sospecha de psitacosis para la búsqueda de C. psittaci. También se estudiaron 4 muestras de aves relacionadas con estos pacientes. De ese total, 48 muestras fueron positivas para C. psittaci mediante una reacción en cadena de la polimerasa (PCR) anidada. Posteriormente, se realizó en el INEI-ANLIS «Dr. Carlos G. Malbrán» la caracterización molecular de 12 muestras positivas para C. psittaci, 8 de humanos y 4 de aves, que fueron genotipificadas por secuenciación del gen ompA. C. psittaci genotipo A se encontró en todas esas muestras. Este informe contribuye a mejorar nuestro conocimiento de las características epidemiológicas y moleculares de C. psittaci para lograr una vigilancia efectiva de la zoonosis que produce." ] ] "NotaPie" => array:1 [ 0 => array:3 [ "etiqueta" => "◊" "nota" => "These authors contributed equally to this article." "identificador" => "fn1" ] ] "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" => 1644 "Ancho" => 2633 "Tamanyo" => 317511 ] ] "descripcion" => array:1 [ "en" => "Neighbor-joining dendrogram based on comparison of 290bp fragment of the ompA gene in Chlamydia. Numbers above branches are bootstrap values as a percentage of 1000 pseudo-replicates and only bootstrap values >60% are shown. Chlamydia caviae GPIC was used as an out-group. Scale bar shows the percentage sequence diversity." ] ] 1 => array:8 [ "identificador" => "tbl0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "detalles" => array:1 [ 0 => array:3 [ "identificador" => "at1" "detalle" => "Table " "rol" => "short" ] ] "tabla" => array:2 [ "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">Patients/birds \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Year \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">City \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Sex \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Age (years) \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Clinical diagnosis \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Sample \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Contact with birds \t\t\t\t\t\t\n \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">67-10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2010 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">San Miguel \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">62 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Community-acquired pneumonia \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Tracheal aspirate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Chicken \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">325-10 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2010 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Buenos Aires \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">45 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Pancreatitis/Community-acquired pneumonia \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">BAL<a class="elsevierStyleCrossRef" href="#tblfn0005">*</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Unknown \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">38-11 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2011 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Santa Rosa \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">38 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Community-acquired pneumonia \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">N&PS<a class="elsevierStyleCrossRef" href="#tblfn0010">a</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Parrot \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Parrot 38/11A \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2011 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Santa Rosa \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">38-13<a class="elsevierStyleCrossRef" href="#tblfn0020">c</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2013 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">San Antonio Oeste \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">48 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Acute respiratory tract infection \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">N&PS<a class="elsevierStyleCrossRef" href="#tblfn0010">a</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Illegal parrot trafficking \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Parrot 35-13A \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2013 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">San Antonio Oeste \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Parrot 38-13A \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2013 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">San Antonio Oeste \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Parrot 135-13A \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2013 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">San Antonio Oeste \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015">b</a> \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">619-13 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2013 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Trenque Lauquen \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">26 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Conjunctivitis \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Ocular swab \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Pigeons \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">357-13 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2013 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Resistencia \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Whooping cough suspected \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">NPA<a class="elsevierStyleCrossRef" href="#tblfn0025">d</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Unknown \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">43-14 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2014 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Neuquén \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">F \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">43 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Acute respiratory tract infection \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">N&PS<a class="elsevierStyleCrossRef" href="#tblfn0010">a</a> \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Parrot \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">83-15 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">2015 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Carmen de Areco \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">M \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">29 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Community-acquired pneumonia \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Tracheal aspirate \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="left" valign="top">Unknown \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab1593275.png" ] ] ] "notaPie" => array:5 [ 0 => array:3 [ "identificador" => "tblfn0005" "etiqueta" => "*" "nota" => "BAL, bronchioalveolar lavage." ] 1 => array:3 [ "identificador" => "tblfn0010" "etiqueta" => "a" "nota" => "N&PS, nasal and pharyngeal swab." ] 2 => array:3 [ "identificador" => "tblfn0015" "etiqueta" => "b" "nota" => "NA, not applicable." ] 3 => array:3 [ "identificador" => "tblfn0020" "etiqueta" => "c" "nota" => "Index case of an outbreak." ] 4 => array:3 [ "identificador" => "tblfn0025" "etiqueta" => "d" "nota" => "NPA, nasopharyngeal aspirate." ] ] ] "descripcion" => array:1 [ "en" => "Characteristics of the C. psittaci-positive cases" ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0005" "bibliografiaReferencia" => array:14 [ 0 => array:3 [ "identificador" => "bib0075" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Genotyping of C. psittaci in central area of Argentina" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:6 [ 0 => "M.C. Frutos" 1 => "M. Monetti" 2 => "X. Kiguen" 3 => "F. Venezuela" 4 => "V. Ré" 5 => "C. Cuffini" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/j.diagmicrobio.2012.06.027" "Revista" => array:6 [ "tituloSerie" => "Diagn Microbiol Infect Dis" "fecha" => "2012" "volumen" => "74" "paginaInicial" => "320" "paginaFinal" => "322" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/22917583" "web" => "Medline" ] ] ] ] ] ] ] ] 1 => array:3 [ "identificador" => "bib0080" "etiqueta" => "2" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Genetic diversity of Chlamydia among captive birds from central Argentina" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:7 [ 0 => "M.C. Frutos" 1 => "M.S. Monetti" 2 => "L. Gallo Vaulet" 3 => "M.E. Cadario" 4 => "M. Rodríguez Fermepin" 5 => "V.E. Ré" 6 => "C.G. 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Características epidemiológicas y moleculares de Chlamydia psittaci provenientes de 8 casos humanos de psitacosis y de 4 aves relacionadas en la Argentina

María E. Cadarioa,◊,

Corresponding author

ecadario@anlis.gov.ar

Corresponding author.

, María C. Frutosb,◊, Maite B. Ariasa, Javier A. Origliac, Vanina Zelayad, María J. Madariagae, Claudia S. Laraa, Viviana Réb, Cecilia G. Cuffinib

a INEI-ANLIS «Dr. Carlos G. Malbrán», Ciudad Autónoma de Buenos Aires, Argentina

b Instituto de Virología «Dr. J.M. Vanella», Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Córdoba, Argentina

c Cátedra de Patología de Aves y Pilíferos, Facultad de Ciencias Veterinarias, Universidad Nacional de La Plata, Argentina

d UOCCB-ANLIS «Dr. Carlos G. Malbrán», Ciudad Autónoma de Buenos Aires, Argentina

e Instituto de Zoonosis Luis Pasteur, Departamento de Diagnóstico y Producción de Biológicos, Ciudad Autónoma de Buenos Aires, Argentina

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    "textoCompleto" => "<span class="elsevierStyleSections"><p id="par0005" class="elsevierStylePara elsevierViewall">Psittacosis is a zoonosis caused by <span class="elsevierStyleItalic">Chlamydia psittaci</span>&#44; an obligate intracellular bacterium belonging to the <span class="elsevierStyleItalic">Chlamydiaceae</span> family and its single genus&#44; <span class="elsevierStyleItalic">Chlamydia</span><a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">13</span></a>&#46;</p><p id="par0010" class="elsevierStylePara elsevierViewall">Human infections vary from unapparent to severe systemic diseases&#58; influenza &#8211; like illnesses&#44; severe atypical pneumonia&#44; endocarditis&#44; myocarditis&#44; meningitis&#44; conjunctivitis&#44; and others&#46; The disease is rarely fatal in properly treated patients&#46; Therefore&#44; awareness of an early diagnosis is important&#46; Pulmonary involvement is common and information of recent exposure to birds is frequently omitted during medical consultation&#46;</p><p id="par0015" class="elsevierStylePara elsevierViewall">Confirmation by molecular methods and genotyping of <span class="elsevierStyleItalic">C&#46; psittaci</span> infection using human respiratory samples and animal samples is a matter of both diagnostic and epidemiological relevance<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">6</span></a>&#46;</p><p id="par0020" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">C&#46; psittaci</span> is divided into at least 9 genotypes &#40;A&#8211;F&#44; E&#47;B&#44; M56 and WC&#41;&#46; Sequence analysis of the <span class="elsevierStyleItalic">ompA</span> gene which encodes the major outer membrane protein &#40;MOMP&#41; gene&#44; is one way to identify all known and eventual new genotypes<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">3</span></a>&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">Genotypes A&#8211;F and E&#47;B are associated with birds and genotypes WC and M56 have been found in cattle and muskrats&#44; respectively<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">3</span></a>&#46; Genotype A is associated with Psittaciformes&#44; B with pigeons&#44; C with ducks and geese<a class="elsevierStyleCrossRef" href="#bib0125"><span class="elsevierStyleSup">11</span></a>&#44; D with turkeys&#44; E with pigeons and ratites&#44; F with psittacine birds and turkeys<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">5</span></a>&#44; and genotype E&#47;B is mainly associated with ducks&#44; turkeys and pigeons<a class="elsevierStyleCrossRefs" href="#bib0085"><span class="elsevierStyleSup">3&#44;7</span></a>&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">The aim of this study was the molecular characterization of <span class="elsevierStyleItalic">C&#46; psittaci</span> polymerase chain reaction-positive &#40;PCR-positive&#41; samples received in the National Reference Laboratory INEI-ANLIS &#8220;Dr&#46; Carlos G&#46; Malbr&#225;n&#8221;&#44; Buenos Aires&#44; Argentina&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">Eight hundred and fifty-six samples from patients of different cities of Argentina&#44; in whom psittacosis was suspected based on clinical presentation and&#47;or history of exposure with infected birds&#44; were analyzed between January 2010 and March 2015&#46; Eight hundred and forty-six were respiratory samples &#40;nasal and pharyngeal swabs&#44; nasopharyngeal aspirates&#44; tracheal aspirates and bronchioalveolar lavages&#41; and 10 were ocular swabs&#46; Cloacal and ocular swabs from 10 related birds were collected and stored until processed&#46; Dacron swabs were placed in a 2<span class="elsevierStyleHsp" style=""></span>ml sucrose&#8211;phosphate&#8211;glutamate or UTM &#40;Copan Italia&#44; Brescia&#44; Italy&#41;&#44; and stored at 4<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; Two hundred microliters of this suspension were subjected to DNA extraction using the DNeasy Blood &#38; Tissue Kit &#40;Qiagen&#44; Valencia&#44; CA&#44; and USA&#41; according to the manufacturer&#39;s instructions&#46; DNA was eluted into 200<span class="elsevierStyleHsp" style=""></span>&#956;l of Qiagen elution buffer and stored at &#8211;20<span class="elsevierStyleHsp" style=""></span>&#176;C until tested&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">A nested PCR protocol was used to amplify a portion of the <span class="elsevierStyleItalic">16S rRNA</span> gene as previously described by Messmer et al&#46;<a class="elsevierStyleCrossRef" href="#bib0120"><span class="elsevierStyleSup">10</span></a> Briefly&#44; 5<span class="elsevierStyleHsp" style=""></span>&#956;l of DNA extract was used in the first step to amplify a 436<span class="elsevierStyleHsp" style=""></span>bp fragment common to the <span class="elsevierStyleItalic">Chlamydia</span> genus&#46; In a second PCR step&#44; 5<span class="elsevierStyleHsp" style=""></span>&#956;l of the PCR product obtained in the previous PCR reaction was used to amplify a 127<span class="elsevierStyleHsp" style=""></span>bp specific product of <span class="elsevierStyleItalic">C&#46; psittaci</span>&#46;</p><p id="par0045" class="elsevierStylePara elsevierViewall">Genotyping of <span class="elsevierStyleItalic">C&#46; psittaci</span> was attempted in all positive samples in the <span class="elsevierStyleItalic">16S rRNA</span> gene PCR&#46; Amplification by nested PCR followed by sequencing of the variable domains III and IV of the <span class="elsevierStyleItalic">ompA</span> gene of <span class="elsevierStyleItalic">C&#46; psittaci</span> was performed as described by Sachse and Hotzel&#46;<a class="elsevierStyleCrossRef" href="#bib0130"><span class="elsevierStyleSup">12</span></a> Briefly&#44; 5<span class="elsevierStyleHsp" style=""></span>&#956;l of DNA extract were used in the first step to amplify a 576<span class="elsevierStyleHsp" style=""></span>bp fragment of the <span class="elsevierStyleItalic">ompA</span> gene common to the <span class="elsevierStyleItalic">Chlamydia</span> genus&#46; In a second PCR step&#44; 5<span class="elsevierStyleHsp" style=""></span>&#956;l of the PCR product obtained in the previous PCR reaction were used to amplify a 389<span class="elsevierStyleHsp" style=""></span>bp specific product of <span class="elsevierStyleItalic">C&#46; psittaci</span>&#46;</p><p id="par0050" class="elsevierStylePara elsevierViewall">Nested PCR products were purified by gel electrophoresis using the AccuPrep&#174; PCR Purification Kit &#40;USA Bioneer Inc&#46;&#41; and subjected to a direct nucleotide sequencing reaction in both directions using the internal &#40;second-round&#41; PCR primers by Macrogen&#44; Inc&#46; &#40;Seoul&#44; Korea&#41;&#46; The sequences obtained from fragments of the <span class="elsevierStyleItalic">ompA</span> gene were edited and prepared with BioEdit V 7&#46;0&#46;9<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">4</span></a> and subsequently aligned with Clustal X 2&#46;12<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">8</span></a>&#44; along with the sequences downloaded from GenBank&#46; Relatedness of newly characterized sequences was assessed by analysis using the 2&#46;2&#46;19 Basic Local Alignment Search Tool&#46; The choice of the genomic region was based on the fact that it is widely associated with the genetic divergence of <span class="elsevierStyleItalic">C&#46; psittaci</span> and it identifies all known genotypes<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">3</span></a>&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">The dendrogram was constructed using the Tree Explorer module of the MEGA software version 4<a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">14</span></a> with the neighbor-joining method and the <span class="elsevierStyleItalic">p</span>-distance parameter&#46; The branch support was evaluated by nonparametric bootstrapping with 1000 pseudo-replicas&#46;</p><p id="par0060" class="elsevierStylePara elsevierViewall">Among the respiratory and ocular swabs samples of the 856 patients studied&#44; 48 &#40;5&#46;6&#37;&#41; tested positive for <span class="elsevierStyleItalic">C&#46; psittaci</span> by nested-PCR&#46; Additionally&#44; 6 samples from related birds tested positive&#46;</p><p id="par0065" class="elsevierStylePara elsevierViewall">Among these 54 positive samples for <span class="elsevierStyleItalic">C&#46; psittaci</span>&#44; only 12 produced enough PCR products in the <span class="elsevierStyleItalic">ompA</span> nested PCR to allow sequencing and genotype determination<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">13</span></a> &#40;8 from humans and 4 from birds&#41;&#46; The mean age of the infected patients was 33&#46;5 years &#40;<span class="elsevierStyleItalic">r</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>5&#8211;62 years&#41;&#59; 62&#46;5&#37; &#40;5&#47;8&#41; were young workers aged 23&#8211;45 years&#46; Male&#47;female ratio was 1&#58;1&#46;</p><p id="par0070" class="elsevierStylePara elsevierViewall">The analysis of the medical records showed that community-acquired pneumonia &#40;CAP&#41; was the most frequent clinical presentation &#40;50&#37;&#59; 4&#47;8&#41;&#44; followed by acute respiratory tract infection &#40;25&#37;&#59; 2&#47;8&#41;&#46; Whooping cough was suspected in one case &#40;12&#46;5&#37;&#59; 1&#47;8&#41;&#46; One patient &#40;12&#46;5&#37;&#59; 1&#47;8&#41; with conjunctivitis tested positive for <span class="elsevierStyleItalic">C&#46; psittaci</span>&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">Five patients reported contact with birds&#44; especially parrots &#40;n<span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>4&#41;&#46; Contact with a domestic or companion bird was not reported in the remaining three <span class="elsevierStyleItalic">C&#46; psittaci</span> positive patients&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">Clinical presentation and sources of the eight <span class="elsevierStyleItalic">C&#46; psittaci</span>-positive samples are shown in <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0085" class="elsevierStylePara elsevierViewall">Genetic diversity and associations among the detected positive samples of <span class="elsevierStyleItalic">C&#46; psittaci</span> were determined by the sequencing and genetic analysis of the composition of the <span class="elsevierStyleItalic">ompA</span> gene&#46; The sequences obtained in this study were deposited in GenBank under accession numbers KU357040&#8211;KU357046&#59; KU357049&#59; KU365350&#8211;KU365353&#46;</p><p id="par0090" class="elsevierStylePara elsevierViewall">In the present study all <span class="elsevierStyleItalic">C&#46; psittaci</span>-positive samples &#40;from humans and birds&#41; were recognized as belonging to genotype A by phylogenetic analysis &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>&#41;&#46; However&#44; in C&#243;rdoba&#44; a central region of Argentina&#44; genotype A was detected in only one case &#40;12&#46;5&#37;&#44; 1&#47;8&#41;&#44; being WC &#40;75&#37;&#44; 6&#47;8&#41; the genotype most frequently found in patients with suspected human psittacosis<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">1</span></a> and in captive birds<a class="elsevierStyleCrossRef" href="#bib0080"><span class="elsevierStyleSup">2</span></a>&#46; Probably the differences could be due to different clinical conditions of the patients in both studies or&#47;and to geographical sources&#46; This differential genotypic profile is an interesting finding that should be investigated in the future&#46; <span class="elsevierStyleItalic">C&#46; psittaci</span> genotype A is more often found among psittacine birds such as parrots and cockatiels&#46; However&#44; the most prevalent <span class="elsevierStyleItalic">C&#46; psittaci</span> genotype in human infections is currently unknown<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">6</span></a>&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0095" class="elsevierStylePara elsevierViewall">The analysis of the medical records showed that CAP was the most frequent clinical presentation&#46; In most studies on CAP&#44; psittacosis was diagnosed only with serological tests&#46; PCR leads to a quick diagnosis and lacks broad genus cross-reactivity&#46; Furthermore&#44; PCR products can be used to genotype <span class="elsevierStyleItalic">C&#46; psittaci</span>&#44; which is relevant for public health notification&#44; source detection and control&#46;</p><p id="par0100" class="elsevierStylePara elsevierViewall">The use of nucleic acid amplification directly from clinical samples and direct genotyping with phylogenetic analysis in the <span class="elsevierStyleItalic">ompA</span> gene could improve the surveillance and molecular epidemiology of this disease&#46; In this sense&#44; the classical method applying two PCR techniques plus sequencing was successfully used and it should be implemented in regional laboratories for an improved diagnosis of this infectious disease&#46;</p><p id="par0105" class="elsevierStylePara elsevierViewall">In this study&#44; a nested PCR and direct sequencing were used to detect and genetically characterize <span class="elsevierStyleItalic">C&#46; psittaci</span>&#46; For the phylogenetic analysis of <span class="elsevierStyleItalic">C&#46; psittaci</span>&#44; we used partial <span class="elsevierStyleItalic">ompA</span> gene amplification&#46; Our results only report the circulation of genotype A associated with respiratory symptoms and conjunctivitis in different areas of our country&#44; at least in our experience with these eight human cases&#46; Psittaciformes especially parakeets and parrots&#44; and individuals in contact with them or with racing pigeons&#44; were the most frequently infected&#46;</p><p id="par0110" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">C&#46; psittaci</span> is rarely suspected in ocular infections&#44; but using molecular techniques&#44; as we did&#44; may be more commonly found<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">9</span></a>&#46; In the present study we showed a case of conjunctivitis due to <span class="elsevierStyleItalic">C&#46; psittaci</span> in a 26 year-old woman not responding to the current treatment and living in contact with wild birds&#46;</p><p id="par0115" class="elsevierStylePara elsevierViewall">Several limitations of our study need to be considered&#46; Isolation in cell cultures was the preferred technique to confirm and corroborate the results obtained&#59; however&#44; in the case of <span class="elsevierStyleItalic">C&#46; psittaci</span>&#44; this procedure is unadvisable because of the biological risks&#46; Moreover&#44; the phylogenetic analysis showed two sequences corresponding to birds &#40;KU365353 and KU365349&#41; in separated clusters&#46; As can be seen in <a class="elsevierStyleCrossRef" href="#fig0005">Figure 1</a> they have a low degree of homology with others obtained in the same city &#40;74&#37; similarity&#44; one with an avian strain and the other with a human strain&#41; &#40;data not shown&#41;&#46; Further studies are needed to confirm the genotype and even the species&#46;</p><p id="par0120" class="elsevierStylePara elsevierViewall">Effective public health surveillance for new variants and modes of transmission will be helpful to understanding the evolution and epidemiology of these bacteria&#46; Therefore&#44; new information about sequence variation of subtypes will be needed&#46;</p><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Ethical responsibilities</span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Protection of human and animal subjects</span><p id="par0125" class="elsevierStylePara elsevierViewall">The authors state that for this investigation no experiments have been performed on humans or animals&#46;</p></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Confidentiality of data</span><p id="par0130" class="elsevierStylePara elsevierViewall">The authors state that they have followed the protocols of their work center on the publication of patient data&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Right to privacy and informed consent</span><p id="par0135" class="elsevierStylePara elsevierViewall">The authors state that in this article there are no patient data&#46;</p></span></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Conflicts of interest</span><p id="par0140" class="elsevierStylePara elsevierViewall">The authors declare that they have no conflicts of interest&#46;</p></span></span>"
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        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">In Argentina&#44; the epidemiological and molecular characteristics of <span class="elsevierStyleItalic">Chlamydia psittaci</span> infections are still not sufficiently known&#46; A total of 846 respiratory and 10 ocular samples from patients with suspected human psittacosis were tested for <span class="elsevierStyleItalic">C&#46; psittaci</span> from January 2010 to March 2015&#46; Four samples of birds related to these patients were also studied&#46; Forty-eight samples were positive for <span class="elsevierStyleItalic">C&#46; psittaci</span> by a nested PCR&#46; The molecular characterization of twelve <span class="elsevierStyleItalic">C&#46; psittaci</span> PCR-positive samples received in the National Reference Laboratory INEI-ANLIS &#8220;Dr&#46; Carlos G&#46; Malbr&#225;n&#8221;&#44; Buenos Aires&#44; Argentina was performed&#46; Eight positive samples from humans and four from birds were genotyped by <span class="elsevierStyleItalic">ompA</span> gene sequencing&#46; <span class="elsevierStyleItalic">C&#46; psittaci</span> genotype A was found in all human samples and in the related birds&#46; This report contributes to our increasing knowledge of the epidemiological and molecular characteristics of <span class="elsevierStyleItalic">C&#46; psittaci</span> to conduct effective surveillance of its zoonotic infections&#46;</p></span>"
      ]
      "es" => array:2 [
        "titulo" => "Resumen"
        "resumen" => "<span id="abst0010" class="elsevierStyleSection elsevierViewall"><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">En la Argentina&#44; a&#250;n no se conocen suficientemente las caracter&#237;sticas epidemiol&#243;gicas y moleculares de las infecciones por <span class="elsevierStyleItalic">Chlamydia psittaci&#46;</span> Entre enero del 2010 y marzo del 2015 se estudiaron 846 muestras respiratorias y 10 oculares de pacientes con sospecha de psitacosis para la b&#250;squeda de <span class="elsevierStyleItalic">C&#46; psittaci</span>&#46; Tambi&#233;n se estudiaron 4 muestras de aves relacionadas con estos pacientes&#46; De ese total&#44; 48 muestras fueron positivas para <span class="elsevierStyleItalic">C&#46; psittaci</span> mediante una reacci&#243;n en cadena de la polimerasa &#40;PCR&#41; anidada&#46; Posteriormente&#44; se realiz&#243; en el INEI-ANLIS &#171;Dr&#46; Carlos G&#46; Malbr&#225;n&#187; la caracterizaci&#243;n molecular de 12 muestras positivas para <span class="elsevierStyleItalic">C&#46; psittaci</span>&#44; 8 de humanos y 4 de aves&#44; que fueron genotipificadas por secuenciaci&#243;n del gen <span class="elsevierStyleItalic">ompA</span>&#46; <span class="elsevierStyleItalic">C&#46; psittaci</span> genotipo A se encontr&#243; en todas esas muestras&#46; Este informe contribuye a mejorar nuestro conocimiento de las caracter&#237;sticas epidemiol&#243;gicas y moleculares de <span class="elsevierStyleItalic">C&#46; psittaci</span> para lograr una vigilancia efectiva de la zoonosis que produce&#46;</p></span>"
      ]
    ]
    "NotaPie" => array:1 [
      0 => array:3 [
        "etiqueta" => "&#9674;"
        "nota" => "<p class="elsevierStyleNotepara" id="npar0030">These authors contributed equally to this article&#46;</p>"
        "identificador" => "fn1"
      ]
    ]
    "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" => 1644
            "Ancho" => 2633
            "Tamanyo" => 317511
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">Neighbor-joining dendrogram based on comparison of 290<span class="elsevierStyleHsp" style=""></span>bp fragment of the <span class="elsevierStyleItalic">ompA</span> gene in <span class="elsevierStyleItalic">Chlamydia</span>&#46; Numbers above branches are bootstrap values as a percentage of 1000 pseudo-replicates and only bootstrap values &#62;60&#37; are shown&#46; <span class="elsevierStyleItalic">Chlamydia caviae</span> GPIC was used as an out-group&#46; Scale bar shows the percentage sequence diversity&#46;</p>"
        ]
      ]
      1 => array:8 [
        "identificador" => "tbl0005"
        "etiqueta" => "Table 1"
        "tipo" => "MULTIMEDIATABLA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "detalles" => array:1 [
          0 => array:3 [
            "identificador" => "at1"
            "detalle" => "Table "
            "rol" => "short"
          ]
        ]
        "tabla" => array:2 [
          "tablatextoimagen" => array:1 [
            0 => array:2 [
              "tabla" => array:1 [
                0 => """
                  <table border="0" frame="\n
                  \t\t\t\t\tvoid\n
                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">Patients&#47;birds&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Year&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">City&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Sex&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Age &#40;years&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Clinical diagnosis&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Sample&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="center" valign="top" scope="col" style="border-bottom: 2px solid black">Contact with birds&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">67-10&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2010&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">San Miguel&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">62&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Community-acquired pneumonia&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Tracheal aspirate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Chicken&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">325-10&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2010&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Buenos Aires&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">45&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Pancreatitis&#47;Community-acquired pneumonia&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">BAL<a class="elsevierStyleCrossRef" href="#tblfn0005"><span class="elsevierStyleSup">&#42;</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Unknown&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">38-11&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2011&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Santa Rosa&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">38&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Community-acquired pneumonia&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">N&#38;PS<a class="elsevierStyleCrossRef" href="#tblfn0010"><span class="elsevierStyleSup">a</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Parrot&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Parrot<br>38&#47;11A&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2011&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Santa Rosa&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">38-13<a class="elsevierStyleCrossRef" href="#tblfn0020"><span class="elsevierStyleSup">c</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2013&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">San Antonio Oeste&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">48&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Acute respiratory tract infection&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">N&#38;PS<a class="elsevierStyleCrossRef" href="#tblfn0010"><span class="elsevierStyleSup">a</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Illegal parrot trafficking&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Parrot<br>35-13A&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2013&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">San Antonio Oeste&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Parrot<br>38-13A&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2013&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">San Antonio Oeste&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Parrot<br>135-13A&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2013&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">San Antonio Oeste&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NA<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">b</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">619-13&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2013&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Trenque Lauquen&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">26&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Conjunctivitis&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Ocular swab&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Pigeons&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">357-13&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2013&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Resistencia&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Whooping cough suspected&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">NPA<a class="elsevierStyleCrossRef" href="#tblfn0025"><span class="elsevierStyleSup">d</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Unknown&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">43-14&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2014&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Neuqu&#233;n&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">F&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">43&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Acute respiratory tract infection&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">N&#38;PS<a class="elsevierStyleCrossRef" href="#tblfn0010"><span class="elsevierStyleSup">a</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Parrot&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">83-15&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">2015&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Carmen de Areco&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">M&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">29&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Community-acquired pneumonia&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Tracheal aspirate&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="left" valign="top">Unknown&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
                  """
              ]
              "imagenFichero" => array:1 [
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Article information

ISSN: 03257541
Original language: English

DOI:10.1016/j.ram.2017.04.001

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2022 August	25	19	44
2022 July	32	23	55
2022 June	45	8	53
2022 May	54	18	72
2022 April	55	14	69
2022 March	125	29	154
2022 February	135	24	159
2022 January	101	37	138
2021 December	80	18	98
2021 November	79	32	111
2021 October	68	39	107
2021 September	59	36	95
2021 August	63	29	92
2021 July	30	11	41
2021 June	25	11	36
2021 May	42	8	50
2021 April	112	16	128
2021 March	41	24	65
2021 February	34	21	55
2021 January	34	24	58
2020 December	28	21	49
2020 November	34	14	48
2020 October	34	7	41
2020 September	34	13	47
2020 August	29	6	35
2020 July	37	3	40
2020 June	38	10	48
2020 May	42	9	51
2020 April	50	7	57
2020 March	53	18	71
2020 February	52	11	63
2020 January	30	3	33
2019 December	34	13	47
2019 November	26	6	32
2019 October	33	3	36
2019 September	62	12	74
2019 August	12	7	19
2019 July	24	3	27
2019 June	79	19	98
2019 May	136	41	177
2019 April	74	32	106
2019 March	15	12	27
2019 February	25	8	33
2019 January	9	7	16
2018 December	16	5	21
2018 November	22	8	30
2018 October	40	11	51
2018 September	10	8	18
2018 August	14	6	20
2018 July	10	3	13
2018 June	14	12	26
2018 May	19	6	25
2018 April	11	3	14
2018 March	13	4	17
2018 February	26	4	30
2018 January	15	2	17
2017 December	24	2	26
2017 November	5	8	13
2017 October	0	8	8
2017 September	1	8	9
2017 August	2	10	12
2017 July	0	5	5

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