Fist report on fumagillin production by Aspergillus fumigatus sensu stricto gliotoxigenic strains recovered from raw cow milk and clinical samples in Argentina

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"ES" => true "ES2" => true "LATM" => true ] "gratuito" => true "lecturas" => array:1 [ "total" => 0 ] "en" => array:13 [ "idiomaDefecto" => true "cabecera" => "Special article" "titulo" => "SARS-CoV-2 and Chlamydia pneumoniae co-infection: A review of the literature" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => array:3 [ 0 => "en" 1 => "en" 2 => "es" ] "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "247" "paginaFinal" => "257" ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Coinfección por Chlamydia pneumoniae y SARS-CoV-2: Una revisión de la literatura" ] ] "contieneResumen" => array:2 [ "en" => true "es" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "María Celia Frutos, Javier Origlia, María Lucia Gallo Vaulet, María Elena Venuta, Miriam Gabriela García, Rita Armitano, Lucía Cipolla, María Julia Madariaga, Cecilia Cuffini, María Estela Cadario" "autores" => array:11 [ 0 => array:2 [ "nombre" => "María Celia" "apellidos" => "Frutos" ] 1 => array:2 [ "nombre" => "Javier" "apellidos" => "Origlia" ] 2 => array:2 [ "nombre" => "María Lucia" "apellidos" => "Gallo Vaulet" ] 3 => array:2 [ "nombre" => "María Elena" "apellidos" => "Venuta" ] 4 => array:2 [ "nombre" => "Miriam Gabriela" "apellidos" => "García" ] 5 => array:2 [ "nombre" => "Rita" "apellidos" => "Armitano" ] 6 => array:2 [ "nombre" => "Lucía" "apellidos" => "Cipolla" ] 7 => array:2 [ "nombre" => "María Julia" "apellidos" => "Madariaga" ] 8 => array:2 [ "nombre" => "Cecilia" "apellidos" => "Cuffini" ] 9 => array:2 [ "nombre" => "María Estela" "apellidos" => "Cadario" ] 10 => array:1 [ "colaborador" => "Grupo de Trabajo Bacterias Atípicas, Sociedad Argentina de Bacteriología, Micología y Parasitología Clínicas (SADEBAC), División de la Asociación Argentina de Microbiología" ] ] ] ] "resumen" => array:1 [ 0 => array:3 [ "titulo" => "Highlights" "clase" => "author-highlights" "resumen" => "<ul class="elsevierStyleList" id="lis0005"><li class="elsevierStyleListItem" id="lsti0005">•Bacterial co-pathogens are commonly identified in viral respiratory infections.</li><li class="elsevierStyleListItem" id="lsti0010">•Co-infection between C. pneumoniae and SARS-CoV-2 has been reported in the literature.</li><li class="elsevierStyleListItem" id="lsti0015">•Information of co-infection in COVID-19 patients is scarce.</li><li class="elsevierStyleListItem" id="lsti0020">•Evidence of co-infections is needed to support developing skills in antimicrobial policy.</li></ul>" ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0325754122000530?idApp=UINPBA00004N" "url" => "/03257541/0000005400000003/v1_202209300808/S0325754122000530/v1_202209300808/en/main.assets" ] "itemAnterior" => array:19 [ "pii" => "S0325754121001024" "issn" => "03257541" "doi" => "10.1016/j.ram.2021.10.005" "estado" => "S300" "fechaPublicacion" => "2022-07-01" "aid" => "479" "copyright" => "Asociación Argentina de Microbiología" "documento" => "article" "crossmark" => 1 "licencia" => "http://creativecommons.org/licenses/by-nc-nd/4.0/" "subdocumento" => "fla" "cita" => "Rev Argent Microbiol. 2022;54:233-42" "abierto" => array:3 [ "ES" => true "ES2" => true "LATM" => true ] "gratuito" => true "lecturas" => array:1 [ "total" => 0 ] "en" => array:14 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Characterization and survival of broad-spectrum biocontrol agents against phytopathogenic fungi" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => array:3 [ 0 => "en" 1 => "en" 2 => "es" ] "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "233" "paginaFinal" => "242" ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Caracterización y supervivencia de agentes de control biológico de amplio espectro frente a hongos fitopatógenos" ] ] "contieneResumen" => array:2 [ "en" => true "es" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 1204 "Ancho" => 2508 "Tamanyo" => 216595 ] ] "descripcion" => array:1 [ "en" => "Graphical representation of the% suppression of growth of phytopathogenic fungi under study using biocontrol bacteria. The y-axis shows the inhibition percentages, while the x-axis represents the bacterial isolates. The bars show the standard error mean computed by using Microsoft Excel." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "Saba Azeem, Syed Inayatullah Agha, Neelam Jamil, Bushra Tabassum, Shan Ahmed, Asif Raheem, Nusrat Jahan, Niaz Ali, Anwar Khan" "autores" => array:9 [ 0 => array:2 [ "nombre" => "Saba" "apellidos" => "Azeem" ] 1 => array:2 [ "nombre" => "Syed Inayatullah" "apellidos" => "Agha" ] 2 => array:2 [ "nombre" => "Neelam" "apellidos" => "Jamil" ] 3 => array:2 [ "nombre" => "Bushra" "apellidos" => "Tabassum" ] 4 => array:2 [ "nombre" => "Shan" "apellidos" => "Ahmed" ] 5 => array:2 [ "nombre" => "Asif" "apellidos" => "Raheem" ] 6 => array:2 [ "nombre" => "Nusrat" "apellidos" => "Jahan" ] 7 => array:2 [ "nombre" => "Niaz" "apellidos" => "Ali" ] 8 => array:2 [ "nombre" => "Anwar" "apellidos" => "Khan" ] ] ] ] "resumen" => array:1 [ 0 => array:3 [ "titulo" => "Highlights" "clase" => "author-highlights" "resumen" => "<ul class="elsevierStyleList" id="lis0005"><li class="elsevierStyleListItem" id="lsti0005">•Bacteria had broad-spectrum activity against phytopathogenic fungi (>50% inhibition).</li><li class="elsevierStyleListItem" id="lsti0010">•Selected isolates were identified with molecular methods and analyzed for plant growth-promoting activities.</li><li class="elsevierStyleListItem" id="lsti0015">•The selected isolates had a good survival rate in a cost-effective medium.</li><li class="elsevierStyleListItem" id="lsti0020">•Assessed medium can be used for commercial fermentation of bio-control agents.</li></ul>" ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0325754121001024?idApp=UINPBA00004N" "url" => "/03257541/0000005400000003/v1_202209300808/S0325754121001024/v1_202209300808/en/main.assets" ] "en" => array:21 [ "idiomaDefecto" => true "cabecera" => "Brief report" "titulo" => "Fist report on fumagillin production by Aspergillus fumigatus sensu stricto gliotoxigenic strains recovered from raw cow milk and clinical samples in Argentina" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "243" "paginaFinal" => "246" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "Verónica A. Alonso, Marina A. Velasco Manini, Gabriela A. Pena, Lilia R. Cavaglieri" "autores" => array:4 [ 0 => array:3 [ "nombre" => "Verónica A." "apellidos" => "Alonso" "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "c" "identificador" => "aff0015" ] ] ] 1 => array:3 [ "nombre" => "Marina A." "apellidos" => "Velasco Manini" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] 2 => array:3 [ "nombre" => "Gabriela A." "apellidos" => "Pena" "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "c" "identificador" => "aff0015" ] ] ] 3 => array:4 [ "nombre" => "Lilia R." "apellidos" => "Cavaglieri" "email" => array:1 [ 0 => "lcavaglieri@exa.unrc.edu.ar" ] "referencia" => array:3 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "c" "identificador" => "aff0015" ] 2 => array:2 [ "etiqueta" => "*" "identificador" => "cor0005" ] ] ] ] "afiliaciones" => array:3 [ 0 => array:3 [ "entidad" => "Departamento de Microbiología e Inmunología, Facultad de Ciencias Exactas, Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Fellow of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina" "etiqueta" => "b" "identificador" => "aff0010" ] 2 => array:3 [ "entidad" => "Member of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina" "etiqueta" => "c" "identificador" => "aff0015" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "Primer reporte en Argentina sobre la producción de fumagilina por cepas de Aspergillus fumigatus sensu stricto productoras de gliotoxina, aisladas de leche cruda y de casos clínicos" ] ] "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" => 1241 "Ancho" => 2500 "Tamanyo" => 117204 ] ] "descripcion" => array:1 [ "en" => "Chromatogram obtained with the injection of 20μl of a 10μg/ml solution of fumagillin." ] ] ] "textoCompleto" => "Aspergillus fumigatus is a worldwide distributed saprophytic fungus and an opportunistic pathogen for human and animals. This fungus is of great importance to public health because it is capable of producing tremorgenic mycotoxins and inducing neurological syndromes in contaminated food handlers, both in maize silage and other contaminated animal feeds as well as in rural workers<a class="elsevierStyleCrossRefs" href="#bib0120">5,8</a>. Aspergillus fumigatus pathogenicity depends not only on the host immune status but also on the virulence of the fungal strain, which is not given by a single factor of essential virulence, but is under polygenetic control. It is known that A. fumigatus produces several immunosuppressive mycotoxins including gliotoxin, fumagillin, helvolic acid, fumitoxins, fumiclavines A and C, fumitremorgins, verruculogen, among others. All these mycotoxins inhibit the function of leukocytes in terms of migration, superoxide production and fungicidal activity<a class="elsevierStyleCrossRef" href="#bib0150">9</a>. Among them, fumagillin has the ability to inhibit endothelial cell proliferation acting as an anti-angiogenic factor, and suppress neutrophil function<a class="elsevierStyleCrossRef" href="#bib0105">2</a>. Our research group has demonstrated that there were no differences between animal and human isolated strains from aspergillosis cases<a class="elsevierStyleCrossRef" href="#bib0140">8</a>. In this way, they may become pathogenic to cows and also to farm handlers. In Argentina, there are currently no reports on the production capacity of fumagillin in native strains. The presence of A. fumigatussensu stricto able to produce other mycotoxins such as fumagillin in raw cow milk could be a very important risk factor since milk and its by-products are destined for human consumption, therefore the aim of this study was to describe the fumagillin production ability of isolated Argentinean A. fumigatussensu stricto strains from raw cow milk and clinical samples.Strains correspond to a broad sampling conducted between March and September 2017; the animals sampled came from 44 dairy establishments located in Córdoba province, in the central region of Argentina. Each one of these establishments owned between 100 and 250 milking cows with daily average milk production of 10–20l/cow. Every dairy establishment was visited once during the morning milking. In each herd, between 20% and 40% of the milking animals (Holando-Argentino breed) were randomly sampled in accordance with the instructions recommended by the National Mastitis Council (2004)<a class="elsevierStyleCrossRef" href="#bib0130">6</a>. A total of 901 individual milking cows were sampled. The udders were disinfected with ethanol 70%, dried with individual paper towel and the first jet of milk discarded. As a result of this sampling, several fungal strains were isolated and identified. Seven of them, corresponding to A. fumigatus isolates were deposited in Universidad Nacional de Río Cuarto, Córdoba, Argentina Collection Center (RC). Three strains isolated from human pulmonary aspergillosis were also added to the collection. All the strains were confirmed as A. fumigatussensu stricto species by morphological and molecular characterization, by sequencing the benA gene of selected strains and by a maximum parsimony analysis<a class="elsevierStyleCrossRef" href="#bib0140">8</a>.Strains were subcultured on malt extract agar (MEA) and incubated at 25°C for 7 days in darkness. Three agar plugs of 6mm diameter (approx. 85mm2 of surface) were removed from different points of the colony to represent its variability, then weighed and introduced into a 250ml Erlenmeyer flask containing 50ml of YES medium (sucrose 150g/l, yeast extract 20g/l modified with 0.5g/l of SO4Mg·7H2O, in distilled water) at 25°C for 7 days.The selected base medium for fumagillin production was developed by Peterson and Goldstein (1957) (saccharose 30g/l, starch 15g/l, NaNO3 3g/l, K2HPO4 1g/l, MgSO4·7H2O 0.5g/l, KCl 0.5g/l, FeSO4·7H2O 0.01g/l) modified by the addition of carboxymethylcellulose (CMC) 10g/l to obtain small pellets<a class="elsevierStyleCrossRef" href="#bib0160">10</a>. Cultures were grown at 30°C for 6 days in an orbital shaker at 220rpm (Heidolph Rotamax 120, Heidolph Instruments GmbH, Schwabach, Germany). A 5ml aliquot of broth was transferred to centrifuge tubes, with 5ml of methanol; the extraction process was carried out at 30°C and 220rpm for 3h. The obtained suspension was centrifuged at 3000×g for 10min, supernatant was filtered through a membrane filter (0.45μm) and dried under nitrogen stream. The extracts were resuspended in 1ml of acetonitrile for HPLC analysis.For obtaining the calibration curve, 1mg pure fumagillin (Sigma USA) was dissolved in 1ml acetonitrile HPLC grade (Sintorgan S.A.). A stock solution of fumagillin 10μg/m and the working solutions of 100, 50, 20, 10, and 5ng/ml were prepared. A Waters e2695 HPLC was used, with a 20μl loading loop and equipped with a diode array detector Waters 2998; separations were carried out on a C18 Phenomenex Luna RP C18 column (150mm×4.6mm, 5μm particle size, Phenomenex Inc, CA, USA) at 25°C. An isocratic mode was used, as the mobile phase, acetonitrile (Sintorgan S.A., Argentina) and water (Sintorgan S.A., Argentina) with 1% acetic acid were used in a 50/50 v/v ratio. The flow for the mobile phase was 1ml/min and the wavelength used was 351nm. Retention time was 15min. The fumagillin concentrations were determined from the areas obtained as a function of the calibration curve. All the strains investigated were able to produce fumagillin at levels between 4.20 and 16.79μg/ml (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>). A chromatogram of the fumagillin peak corresponding to the injection of 20μl of pure standard (10μg/ml) can be observed at 14.5min (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>). The analyzed strains from raw cow milk samples showed a variable production capacity of fumagillin. The strain A. fumigatus RC2243, from a cow milk sample with clinical mastitis, showed the highest production capacity among all the tested strains. Strains from healthy cow samples yielded intermediate values between 9.23 and 9.97μg/ml, while the lowest level was shown in A. fumigatus RC2251 strain, isolated from a subclinical mastitis sample. With the purpose of comparing the toxic potential of strains from different origins, three strains of A. fumigatussensu stricto from human pulmonary aspergillosis were included in the study. Among these strains, one of them had the highest production capacity (12.74μg/ml) while the others had moderate fumagillin production capacity. Pellegrino et al. (2013)<a class="elsevierStyleCrossRef" href="#bib0135">7</a> conducted a mycological survey of the milk samples from which the strains of the current study were isolated and found that A. fumigatus was the second more frequently isolated species of Aspergillus after A. flavus. Moreover, they demonstrated the gliotoxinogenic ability of the strains and its association with high somatic cell counts in milk samples. The results obtained in the present work constitute a first step in the knowledge of the production capacity of fumagillin from A. fumigatussensu stricto gliotoxin producer strains obtained from animal and human samples in Argentina. Boudra and Morgavi (2005)<a class="elsevierStyleCrossRef" href="#bib0095">1</a> analyzed the toxicogenic capacity of A. fumigatus strains in different natural feed substrates (wheat, corn, barley, triticale, rye grass, orchard grass, and tall fescue); fumagillin production levels up to 25.9μg/ml were obtained after six days of development in a medium other than that used in the present study (minimal essential medium Eagle supplemented with fetal bovine serum and glucose). In another work, the growth and production of fumagillin by A. fumigatus NRRL 2436 was measured in Peterson-Goldstein medium modified with carboxymethylcellulose and similar fumagillin levels to ours were obtained (average of 13.4mg/l)<a class="elsevierStyleCrossRef" href="#bib0160">10</a>. The medium used in the present work was useful to perform a screening among strains in order to identify those with a capacity to produce fumagillin. This knowledge is relevant due to the well-known antiangiogenic effects of fumagillin that reduce the proliferation of endothelial cells and blood vessel formation<a class="elsevierStyleCrossRef" href="#bib0115">4</a> probably related to the process of A. fumigatus pathogenesis in invasive aspergillosis. Guruceaga et al. (2018)<a class="elsevierStyleCrossRef" href="#bib0110">3</a> showed the gene-expression and metabolite bioactivity of the fumagillin/pseurotin gene cluster during invasive lung aspergillosis after intranasal exposure to A. fumigatus conidia using a murine model. They concluded that fumagillin contributed to tissue damage during invasive aspergillosis, and allowed A. fumigatus progress by the lungs through angioinvasion and disruption of the lung parenchymal structure. Furthermore, it has been shown that fumagillin inhibits neutrophil function contributing to the reduction of the local immune response<a class="elsevierStyleCrossRef" href="#bib0105">2</a>. These facts confirm or increase the risk posed by the workers’ exposure to fumagillin contamination. Although fumagillin toxicity to humans is important because it poses a direct risk to the consumer, this mycotoxin is on the list of products approved for use in beekeeping in several countries including Argentina, according to SENASA regulations (Cert. No. 97.117) for the treatment of nosema disease (Nosema apis and Nosema ceranae) and is commonly used. In the present work, all the strains tested for the production of fumagillin were able to synthesize it, highlighting that the strain A. fumigatus RC2243, from a milk sample of a cow with clinical mastitis, was the most productive. It should be noted that the strains isolated and studied in this work came from the animal environment, specifically the dairy farm environment, all of which are able to produce at least two of the most important mycotoxins that have been identified in A. fumigatus. This increases the possibility of being transferred to milk and polluting the environment in which they develop, constituting a public health risk. It is necessary to conduct specific trials for investigating the efficiency of different mycotoxin adsorbents and biodegradation products against several toxins that are mainly detected in forage and avoid its transfer to the human food chain.<elsevierMultimedia ident="tbl0005"></elsevierMultimedia><elsevierMultimedia ident="fig0005"></elsevierMultimedia>Conflict of interestThe authors declare that they have no conflicts of interest." "textoCompletoSecciones" => array:1 [ "secciones" => array:8 [ 0 => array:3 [ "identificador" => "xres1777058" "titulo" => "Highlights" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0005" ] ] ] 1 => array:3 [ "identificador" => "xres1777057" "titulo" => "Abstract" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0010" ] ] ] 2 => array:2 [ "identificador" => "xpalclavsec1560259" "titulo" => "Keywords" ] 3 => array:3 [ "identificador" => "xres1777056" "titulo" => "Resumen" "secciones" => array:1 [ 0 => array:1 [ "identificador" => "abst0015" ] ] ] 4 => array:2 [ "identificador" => "xpalclavsec1560260" "titulo" => "Palabras clave" ] 5 => array:2 [ "identificador" => "sec0005" "titulo" => "Conflict of interest" ] 6 => array:2 [ "identificador" => "xack627968" "titulo" => "Acknowledgements" ] 7 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2020-06-10" "fechaAceptado" => "2022-03-04" "PalabrasClave" => array:2 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec1560259" "palabras" => array:4 [ 0 => "Fumagillin" 1 => "Aspergillus fumigatus" 2 => "Raw cow milk strains" 3 => "Clinical strains" ] ] ] "es" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Palabras clave" "identificador" => "xpalclavsec1560260" "palabras" => array:4 [ 0 => "Fumagillina" 1 => "Aspergillus fumigatus" 2 => "Cepas aisladas de leche cruda" 3 => "Cepas clínicas" ] ] ] ] "tieneResumen" => true "highlights" => array:2 [ "titulo" => "Highlights" "resumen" => "<ul class="elsevierStyleList" id="lis0005"><li class="elsevierStyleListItem" id="lsti0005">•A. fumigatus strains from farm and clinical samples were able to produce fumagillin.</li><li class="elsevierStyleListItem" id="lsti0010">•A. fumigatus strains were gliotoxin and fumagillin producers.</li><li class="elsevierStyleListItem" id="lsti0015">•Fumagillin producers represent public health risk of toxin transference to raw milk.</li></ul>" ] "resumen" => array:2 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "In Argentina there are no reports on Aspergillus fumigatus fumagillin-producing strains. In this study we describe the isolation and mycotoxin production capacity of ten A. fumigatus strains isolated from farm and clinical samples. Farm strains were isolated from milk samples taken from dairy cows in Córdoba province, some of which were associated with subclinical mastitis. A culture medium was defined to optimize fumagillin production and a detection method was developed by HPLC chromatography. It is known that in addition to the host immune status, strain virulence is a fundamental characteristic that will determine its pathogenicity and, in this sense, fumagillin is considered to be among the virulence factors. In the present work, all the strains tested for the production of fumagillin were able to synthesize it, highlighting that the strain A. fumigatus RC2243, from a milk sample from a cow with clinical mastitis, was the most productive. The existence of fumagillin-producing strains represents a potential risk of mycotoxins being transferred to raw milk, constituting a public health risk." ] "es" => array:2 [ "titulo" => "Resumen" "resumen" => "En Argentina no existen reportes sobre cepas de Aspergillus fumigatus productoras de fumagilina. En este trabajo se describe el aislamiento y la producción de dicha micotoxina por 10 cepas, provenientes del medioambiente rural y aisladas de muestras clínicas. Las cepas de origen rural fueron aisladas de vacas lecheras en tambos de la provincia de Córdoba, y algunas de esas cepas se asociaron a casos de mastitis subclínica. Se definió la composición de un medio de cultivo para optimizar la producción de fumagilina y se desarrolló un método de cromatografía HPLC para su determinación. Es conocido que, además del estado inmunitario del huésped, la virulencia de la cepa es una de las características fundamentales que determinan su potencial patogénico y, en este sentido, la fumagilina es considerada un factor de virulencia. En el presente trabajo todas las cepas estudiadas fueron capaces de sintetizarla y la cepa A. fumigatus RC2243, proveniente de leche de una vaca con mastitis subclínica, se destacó como la cepa más productora. La existencia de cepas productoras de fumagillina representa un riesgo potencial por el pasaje de dicha micotoxina a la leche, lo cual constituye un problema para la salud pública." ] ] "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" => 1241 "Ancho" => 2500 "Tamanyo" => 117204 ] ] "descripcion" => array:1 [ "en" => "Chromatogram obtained with the injection of 20μl of a 10μg/ml solution of fumagillin." ] ] 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:1 [ "tablatextoimagen" => array:1 [ 0 => array:1 [ "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="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Origin \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Strain \t\t\t\t\t\t\n \t\t\t\t\t\t</th><th class="td" title="\n \t\t\t\t\ttable-head\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Fumagillin (μg·ml−1) \t\t\t\t\t\t\n \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="7" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Cow milk</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2210 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">9.23 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2213 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">9.23 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2229 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">9.97 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2240 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5.4 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2243 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">16.79 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2249 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">5.47 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2251 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">4.2 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " rowspan="3" align="left" valign="\n \t\t\t\t\ttop\n \t\t\t\t">Human aspergillosis</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2257* \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">8.93 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2261* \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">12.74 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">RC2263* \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="\n \t\t\t\t\ttable-entry\n \t\t\t\t " align="center" valign="\n \t\t\t\t\ttop\n \t\t\t\t">7.97 \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] ] ] ] "descripcion" => array:1 [ "en" => "Fumagillin concentration produced by A. fumigatus strains in Peterson–Goldstein modified medium." ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0015" "bibliografiaReferencia" => array:10 [ 0 => array:3 [ "identificador" => "bib0095" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Mycotoxin risk evaluation in feeds contaminated by Aspergillus fumigatus" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "H. 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Garaizar" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:5 [ "tituloSerie" => "Rev Iberoam de Micol" "fecha" => "2005" "volumen" => "22" "paginaInicial" => "1" "paginaFinal" => "23" ] ] ] ] ] ] 9 => array:3 [ "identificador" => "bib0160" "etiqueta" => "10" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Carbon and nitrogen source nutrition of fumagillin biosynthesis by Aspergillus fumigatus" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:4 [ 0 => "W. Yang" 1 => "W.S. Kim" 2 => "A. Fang" 3 => "A.L. Demain" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1007/s00284-002-3842-2" "Revista" => array:6 [ "tituloSerie" => "Curr Microbiol" "fecha" => "2003" "volumen" => "46" "paginaInicial" => "275" "paginaFinal" => "279" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/12732977" "web" => "Medline" ] ] ] ] ] ] ] ] ] ] ] ] "agradecimientos" => array:1 [ 0 => array:4 [ "identificador" => "xack627968" "titulo" => "Acknowledgements" "texto" => "This work was supported by grants from FONCyT-PICT2033/15." "vista" => "all" ] ] ] "idiomaDefecto" => "en" "url" => "/03257541/0000005400000003/v1_202209300808/S0325754122000219/v1_202209300808/en/main.assets" "Apartado" => array:4 [ "identificador" => "37862" "tipo" => "SECCION" "en" => array:2 [ "titulo" => "Microbiología agrícola, ambiental e industrial" "idiomaDefecto" => true ] "idiomaDefecto" => "en" ] "PDF" => "https://static.elsevier.es/multimedia/03257541/0000005400000003/v1_202209300808/S0325754122000219/v1_202209300808/en/main.pdf?idApp=UINPBA00004N&text.app=https://www.elsevier.es/" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0325754122000219?idApp=UINPBA00004N"]

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Primer reporte en Argentina sobre la producción de fumagilina por cepas de Aspergillus fumigatus sensu stricto productoras de gliotoxina, aisladas de leche cruda y de casos clínicos

Verónica A. Alonsoa,c, Marina A. Velasco Maninib, Gabriela A. Penaa,c, Lilia R. Cavaglieria,c,

Corresponding author

lcavaglieri@exa.unrc.edu.ar

Corresponding author.

a Departamento de Microbiología e Inmunología, Facultad de Ciencias Exactas, Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina

b Fellow of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina

c Member of Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina

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          "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall"><ul class="elsevierStyleList" id="lis0005"><li class="elsevierStyleListItem" id="lsti0005"><span class="elsevierStyleLabel">&#8226;</span><p id="par0005" class="elsevierStylePara elsevierViewall">Bacterial co-pathogens are commonly identified in viral respiratory infections&#46;</p></li><li class="elsevierStyleListItem" id="lsti0010"><span class="elsevierStyleLabel">&#8226;</span><p id="par0010" class="elsevierStylePara elsevierViewall">Co-infection between <span class="elsevierStyleItalic">C&#46; pneumoniae</span> and SARS-CoV-2 has been reported in the literature&#46;</p></li><li class="elsevierStyleListItem" id="lsti0015"><span class="elsevierStyleLabel">&#8226;</span><p id="par0015" class="elsevierStylePara elsevierViewall">Information of co-infection in COVID-19 patients is scarce&#46;</p></li><li class="elsevierStyleListItem" id="lsti0020"><span class="elsevierStyleLabel">&#8226;</span><p id="par0020" class="elsevierStylePara elsevierViewall">Evidence of co-infections is needed to support developing skills in antimicrobial policy&#46;</p></li></ul></p></span>"
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    "doi" => "10.1016/j.ram.2021.10.005"
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          "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall"><ul class="elsevierStyleList" id="lis0005"><li class="elsevierStyleListItem" id="lsti0005"><span class="elsevierStyleLabel">&#8226;</span><p id="par0005" class="elsevierStylePara elsevierViewall">Bacteria had broad-spectrum activity against phytopathogenic fungi &#40;&#62;50&#37; inhibition&#41;&#46;</p></li><li class="elsevierStyleListItem" id="lsti0010"><span class="elsevierStyleLabel">&#8226;</span><p id="par0010" class="elsevierStylePara elsevierViewall">Selected isolates were identified with molecular methods and analyzed for plant growth-promoting activities&#46;</p></li><li class="elsevierStyleListItem" id="lsti0015"><span class="elsevierStyleLabel">&#8226;</span><p id="par0015" class="elsevierStylePara elsevierViewall">The selected isolates had a good survival rate in a cost-effective medium&#46;</p></li><li class="elsevierStyleListItem" id="lsti0020"><span class="elsevierStyleLabel">&#8226;</span><p id="par0020" class="elsevierStylePara elsevierViewall">Assessed medium can be used for commercial fermentation of bio-control agents&#46;</p></li></ul></p></span>"
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    "cabecera" => "<span class="elsevierStyleTextfn">Brief report</span>"
    "titulo" => "Fist report on fumagillin production by <span class="elsevierStyleItalic">Aspergillus fumigatus sensu stricto</span> gliotoxigenic strains recovered from raw cow milk and clinical samples in Argentina"
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        "titulo" => "Primer reporte en Argentina sobre la producci&#243;n de fumagilina por cepas de <span class="elsevierStyleItalic">Aspergillus fumigatus sensu stricto</span> productoras de gliotoxina&#44; aisladas de leche cruda y de casos cl&#237;nicos"
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          "en" => "<p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">Chromatogram obtained with the injection of 20<span class="elsevierStyleHsp" style=""></span>&#956;l of a 10<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml solution of fumagillin&#46;</p>"
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    "textoCompleto" => "<span class="elsevierStyleSections"><p id="par0020" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">Aspergillus fumigatus</span> is a worldwide distributed saprophytic fungus and an opportunistic pathogen for human and animals&#46; This fungus is of great importance to public health because it is capable of producing tremorgenic mycotoxins and inducing neurological syndromes in contaminated food handlers&#44; both in maize silage and other contaminated animal feeds as well as in rural workers<a class="elsevierStyleCrossRefs" href="#bib0120"><span class="elsevierStyleSup">5&#44;8</span></a>&#46; <span class="elsevierStyleItalic">Aspergillus fumigatus</span> pathogenicity depends not only on the host immune status but also on the virulence of the fungal strain&#44; which is not given by a single factor of essential virulence&#44; but is under polygenetic control&#46; It is known that <span class="elsevierStyleItalic">A&#46; fumigatus</span> produces several immunosuppressive mycotoxins including gliotoxin&#44; fumagillin&#44; helvolic acid&#44; fumitoxins&#44; fumiclavines A and C&#44; fumitremorgins&#44; verruculogen&#44; among others&#46; All these mycotoxins inhibit the function of leukocytes in terms of migration&#44; superoxide production and fungicidal activity<a class="elsevierStyleCrossRef" href="#bib0150"><span class="elsevierStyleSup">9</span></a>&#46; Among them&#44; fumagillin has the ability to inhibit endothelial cell proliferation acting as an anti-angiogenic factor&#44; and suppress neutrophil function<a class="elsevierStyleCrossRef" href="#bib0105"><span class="elsevierStyleSup">2</span></a>&#46; Our research group has demonstrated that there were no differences between animal and human isolated strains from aspergillosis cases<a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">8</span></a>&#46; In this way&#44; they may become pathogenic to cows and also to farm handlers&#46; In Argentina&#44; there are currently no reports on the production capacity of fumagillin in native strains&#46; The presence of <span class="elsevierStyleItalic">A&#46; fumigatus</span><span class="elsevierStyleItalic">sensu stricto</span> able to produce other mycotoxins such as fumagillin in raw cow milk could be a very important risk factor since milk and its by-products are destined for human consumption&#44; therefore the aim of this study was to describe the fumagillin production ability of isolated Argentinean <span class="elsevierStyleItalic">A&#46; fumigatus</span><span class="elsevierStyleItalic">sensu stricto</span> strains from raw cow milk and clinical samples&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">Strains correspond to a broad sampling conducted between March and September 2017&#59; the animals sampled came from 44 dairy establishments located in C&#243;rdoba province&#44; in the central region of Argentina&#46; Each one of these establishments owned between 100 and 250 milking cows with daily average milk production of 10&#8211;20<span class="elsevierStyleHsp" style=""></span>l&#47;cow&#46; Every dairy establishment was visited once during the morning milking&#46; In each herd&#44; between 20&#37; and 40&#37; of the milking animals &#40;Holando-Argentino breed&#41; were randomly sampled in accordance with the instructions recommended by the National Mastitis Council &#40;2004&#41;<a class="elsevierStyleCrossRef" href="#bib0130"><span class="elsevierStyleSup">6</span></a>&#46; A total of 901 individual milking cows were sampled&#46; The udders were disinfected with ethanol 70&#37;&#44; dried with individual paper towel and the first jet of milk discarded&#46; As a result of this sampling&#44; several fungal strains were isolated and identified&#46; Seven of them&#44; corresponding to <span class="elsevierStyleItalic">A&#46; fumigatus</span> isolates were deposited in Universidad Nacional de R&#237;o Cuarto&#44; C&#243;rdoba&#44; Argentina Collection Center &#40;RC&#41;&#46; Three strains isolated from human pulmonary aspergillosis were also added to the collection&#46; All the strains were confirmed as <span class="elsevierStyleItalic">A&#46; fumigatus</span><span class="elsevierStyleItalic">sensu stricto</span> species by morphological and molecular characterization&#44; by sequencing the benA gene of selected strains and by a maximum parsimony analysis<a class="elsevierStyleCrossRef" href="#bib0140"><span class="elsevierStyleSup">8</span></a>&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">Strains were subcultured on malt extract agar &#40;MEA&#41; and incubated at 25<span class="elsevierStyleHsp" style=""></span>&#176;C for 7 days in darkness&#46; Three agar plugs of 6<span class="elsevierStyleHsp" style=""></span>mm diameter &#40;approx&#46; 85<span class="elsevierStyleHsp" style=""></span>mm<span class="elsevierStyleSup">2</span> of surface&#41; were removed from different points of the colony to represent its variability&#44; then weighed and introduced into a 250<span class="elsevierStyleHsp" style=""></span>ml Erlenmeyer flask containing 50<span class="elsevierStyleHsp" style=""></span>ml of YES medium &#40;sucrose 150<span class="elsevierStyleHsp" style=""></span>g&#47;l&#44; yeast extract 20<span class="elsevierStyleHsp" style=""></span>g&#47;l modified with 0&#46;5<span class="elsevierStyleHsp" style=""></span>g&#47;l of SO<span class="elsevierStyleInf">4</span>Mg&#183;7H<span class="elsevierStyleInf">2</span>O&#44; in distilled water&#41; at 25<span class="elsevierStyleHsp" style=""></span>&#176;C for 7 days&#46;</p><p id="par0035" class="elsevierStylePara elsevierViewall">The selected base medium for fumagillin production was developed by Peterson and Goldstein &#40;1957&#41; &#40;saccharose 30<span class="elsevierStyleHsp" style=""></span>g&#47;l&#44; starch 15<span class="elsevierStyleHsp" style=""></span>g&#47;l&#44; NaNO<span class="elsevierStyleInf">3</span> 3<span class="elsevierStyleHsp" style=""></span>g&#47;l&#44; K<span class="elsevierStyleInf">2</span>HPO<span class="elsevierStyleInf">4</span> 1<span class="elsevierStyleHsp" style=""></span>g&#47;l&#44; MgSO<span class="elsevierStyleInf">4</span>&#183;7H<span class="elsevierStyleInf">2</span>O 0&#46;5<span class="elsevierStyleHsp" style=""></span>g&#47;l&#44; KCl 0&#46;5<span class="elsevierStyleHsp" style=""></span>g&#47;l&#44; FeSO<span class="elsevierStyleInf">4</span>&#183;7H<span class="elsevierStyleInf">2</span>O 0&#46;01<span class="elsevierStyleHsp" style=""></span>g&#47;l&#41; modified by the addition of carboxymethylcellulose &#40;CMC&#41; 10<span class="elsevierStyleHsp" style=""></span>g&#47;l to obtain small pellets<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">10</span></a>&#46; Cultures were grown at 30<span class="elsevierStyleHsp" style=""></span>&#176;C for 6 days in an orbital shaker at 220<span class="elsevierStyleHsp" style=""></span>rpm &#40;Heidolph Rotamax 120&#44; Heidolph Instruments GmbH&#44; Schwabach&#44; Germany&#41;&#46; A 5<span class="elsevierStyleHsp" style=""></span>ml aliquot of broth was transferred to centrifuge tubes&#44; with 5<span class="elsevierStyleHsp" style=""></span>ml of methanol&#59; the extraction process was carried out at 30<span class="elsevierStyleHsp" style=""></span>&#176;C and 220<span class="elsevierStyleHsp" style=""></span>rpm for 3<span class="elsevierStyleHsp" style=""></span>h&#46; The obtained suspension was centrifuged at 3000<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span> for 10<span class="elsevierStyleHsp" style=""></span>min&#44; supernatant was filtered through a membrane filter &#40;0&#46;45<span class="elsevierStyleHsp" style=""></span>&#956;m&#41; and dried under nitrogen stream&#46; The extracts were resuspended in 1<span class="elsevierStyleHsp" style=""></span>ml of acetonitrile for HPLC analysis&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">For obtaining the calibration curve&#44; 1<span class="elsevierStyleHsp" style=""></span>mg pure fumagillin &#40;Sigma USA&#41; was dissolved in 1<span class="elsevierStyleHsp" style=""></span>ml acetonitrile HPLC grade &#40;Sintorgan S&#46;A&#46;&#41;&#46; A stock solution of fumagillin 10<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;m and the working solutions of 100&#44; 50&#44; 20&#44; 10&#44; and 5<span class="elsevierStyleHsp" style=""></span>ng&#47;ml were prepared&#46; A Waters e2695 HPLC was used&#44; with a 20<span class="elsevierStyleHsp" style=""></span>&#956;l loading loop and equipped with a diode array detector Waters 2998&#59; separations were carried out on a C18 Phenomenex Luna RP C18 column &#40;150<span class="elsevierStyleHsp" style=""></span>mm<span class="elsevierStyleHsp" style=""></span>&#215;<span class="elsevierStyleHsp" style=""></span>4&#46;6<span class="elsevierStyleHsp" style=""></span>mm&#44; 5<span class="elsevierStyleHsp" style=""></span>&#956;m particle size&#44; Phenomenex Inc&#44; CA&#44; USA&#41; at 25<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; An isocratic mode was used&#44; as the mobile phase&#44; acetonitrile &#40;Sintorgan S&#46;A&#46;&#44; Argentina&#41; and water &#40;Sintorgan S&#46;A&#46;&#44; Argentina&#41; with 1&#37; acetic acid were used in a 50&#47;50 v&#47;v ratio&#46; The flow for the mobile phase was 1<span class="elsevierStyleHsp" style=""></span>ml&#47;min and the wavelength used was 351<span class="elsevierStyleHsp" style=""></span>nm&#46; Retention time was 15<span class="elsevierStyleHsp" style=""></span>min&#46; The fumagillin concentrations were determined from the areas obtained as a function of the calibration curve&#46; All the strains investigated were able to produce fumagillin at levels between 4&#46;20 and 16&#46;79<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml &#40;<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#41;&#46; A chromatogram of the fumagillin peak corresponding to the injection of 20<span class="elsevierStyleHsp" style=""></span>&#956;l of pure standard &#40;10<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#41; can be observed at 14&#46;5<span class="elsevierStyleHsp" style=""></span>min &#40;<a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>&#41;&#46; The analyzed strains from raw cow milk samples showed a variable production capacity of fumagillin&#46; The strain <span class="elsevierStyleItalic">A&#46; fumigatus</span> RC2243&#44; from a cow milk sample with clinical mastitis&#44; showed the highest production capacity among all the tested strains&#46; Strains from healthy cow samples yielded intermediate values between 9&#46;23 and 9&#46;97<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#44; while the lowest level was shown in <span class="elsevierStyleItalic">A&#46; fumigatus</span> RC2251 strain&#44; isolated from a subclinical mastitis sample&#46; With the purpose of comparing the toxic potential of strains from different origins&#44; three strains of <span class="elsevierStyleItalic">A&#46; fumigatus</span><span class="elsevierStyleItalic">sensu stricto</span> from human pulmonary aspergillosis were included in the study&#46; Among these strains&#44; one of them had the highest production capacity &#40;12&#46;74<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#41; while the others had moderate fumagillin production capacity&#46; Pellegrino et al&#46; &#40;2013&#41;<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">7</span></a> conducted a mycological survey of the milk samples from which the strains of the current study were isolated and found that <span class="elsevierStyleItalic">A&#46; fumigatus</span> was the second more frequently isolated species of <span class="elsevierStyleItalic">Aspergillus</span> after <span class="elsevierStyleItalic">A&#46; flavus</span>&#46; Moreover&#44; they demonstrated the gliotoxinogenic ability of the strains and its association with high somatic cell counts in milk samples&#46; The results obtained in the present work constitute a first step in the knowledge of the production capacity of fumagillin from <span class="elsevierStyleItalic">A&#46; fumigatus</span><span class="elsevierStyleItalic">sensu stricto</span> gliotoxin producer strains obtained from animal and human samples in Argentina&#46; Boudra and Morgavi &#40;2005&#41;<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">1</span></a> analyzed the toxicogenic capacity of <span class="elsevierStyleItalic">A&#46; fumigatus</span> strains in different natural feed substrates &#40;wheat&#44; corn&#44; barley&#44; triticale&#44; rye grass&#44; orchard grass&#44; and tall fescue&#41;&#59; fumagillin production levels up to 25&#46;9<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml were obtained after six days of development in a medium other than that used in the present study &#40;minimal essential medium Eagle supplemented with fetal bovine serum and glucose&#41;&#46; In another work&#44; the growth and production of fumagillin by <span class="elsevierStyleItalic">A&#46; fumigatus</span> NRRL 2436 was measured in Peterson-Goldstein medium modified with carboxymethylcellulose and similar fumagillin levels to ours were obtained &#40;average of 13&#46;4<span class="elsevierStyleHsp" style=""></span>mg&#47;l&#41;<a class="elsevierStyleCrossRef" href="#bib0160"><span class="elsevierStyleSup">10</span></a>&#46; The medium used in the present work was useful to perform a screening among strains in order to identify those with a capacity to produce fumagillin&#46; This knowledge is relevant due to the well-known antiangiogenic effects of fumagillin that reduce the proliferation of endothelial cells and blood vessel formation<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">4</span></a> probably related to the process of <span class="elsevierStyleItalic">A&#46; fumigatus</span> pathogenesis in invasive aspergillosis&#46; Guruceaga et al&#46; &#40;2018&#41;<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">3</span></a> showed the gene-expression and metabolite bioactivity of the fumagillin&#47;pseurotin gene cluster during invasive lung aspergillosis after intranasal exposure to <span class="elsevierStyleItalic">A&#46; fumigatus</span> conidia using a murine model&#46; They concluded that fumagillin contributed to tissue damage during invasive aspergillosis&#44; and allowed <span class="elsevierStyleItalic">A&#46; fumigatus</span> progress by the lungs through angioinvasion and disruption of the lung parenchymal structure&#46; Furthermore&#44; it has been shown that fumagillin inhibits neutrophil function contributing to the reduction of the local immune response<a class="elsevierStyleCrossRef" href="#bib0105"><span class="elsevierStyleSup">2</span></a>&#46; These facts confirm or increase the risk posed by the workers&#8217; exposure to fumagillin contamination&#46; Although fumagillin toxicity to humans is important because it poses a direct risk to the consumer&#44; this mycotoxin is on the list of products approved for use in beekeeping in several countries including Argentina&#44; according to SENASA regulations &#40;Cert&#46; No&#46; 97&#46;117&#41; for the treatment of nosema disease &#40;<span class="elsevierStyleItalic">Nosema apis</span> and <span class="elsevierStyleItalic">Nosema ceranae</span>&#41; and is commonly used&#46; In the present work&#44; all the strains tested for the production of fumagillin were able to synthesize it&#44; highlighting that the strain <span class="elsevierStyleItalic">A&#46; fumigatus</span> RC2243&#44; from a milk sample of a cow with clinical mastitis&#44; was the most productive&#46; It should be noted that the strains isolated and studied in this work came from the animal environment&#44; specifically the dairy farm environment&#44; all of which are able to produce at least two of the most important mycotoxins that have been identified in <span class="elsevierStyleItalic">A&#46; fumigatus</span>&#46; This increases the possibility of being transferred to milk and polluting the environment in which they develop&#44; constituting a public health risk&#46; It is necessary to conduct specific trials for investigating the efficiency of different mycotoxin adsorbents and biodegradation products against several toxins that are mainly detected in forage and avoid its transfer to the human food chain&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><elsevierMultimedia ident="fig0005"></elsevierMultimedia><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Conflict of interest</span><p id="par0045" class="elsevierStylePara elsevierViewall">The authors declare that they have no conflicts of interest&#46;</p></span></span>"
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          "titulo" => "Conflict of interest"
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          "identificador" => "xack627968"
          "titulo" => "Acknowledgements"
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          "titulo" => "References"
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    "fechaRecibido" => "2020-06-10"
    "fechaAceptado" => "2022-03-04"
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            0 => "Fumagillin"
            1 => "<span class="elsevierStyleItalic">Aspergillus fumigatus</span>"
            2 => "Raw cow milk strains"
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          "palabras" => array:4 [
            0 => "Fumagillina"
            1 => "<span class="elsevierStyleItalic">Aspergillus fumigatus</span>"
            2 => "Cepas aisladas de leche cruda"
            3 => "Cepas cl&#237;nicas"
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    "highlights" => array:2 [
      "titulo" => "Highlights"
      "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall"><ul class="elsevierStyleList" id="lis0005"><li class="elsevierStyleListItem" id="lsti0005"><span class="elsevierStyleLabel">&#8226;</span><p id="par0005" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">A&#46; fumigatus</span> strains from farm and clinical samples were able to produce fumagillin&#46;</p></li><li class="elsevierStyleListItem" id="lsti0010"><span class="elsevierStyleLabel">&#8226;</span><p id="par0010" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">A&#46; fumigatus</span> strains were gliotoxin and fumagillin producers&#46;</p></li><li class="elsevierStyleListItem" id="lsti0015"><span class="elsevierStyleLabel">&#8226;</span><p id="par0015" class="elsevierStylePara elsevierViewall">Fumagillin producers represent public health risk of toxin transference to raw milk&#46;</p></li></ul></p></span>"
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    "resumen" => array:2 [
      "en" => array:2 [
        "titulo" => "Abstract"
        "resumen" => "<span id="abst0010" class="elsevierStyleSection elsevierViewall"><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">In Argentina there are no reports on <span class="elsevierStyleItalic">Aspergillus fumigatus</span> fumagillin-producing strains&#46; In this study we describe the isolation and mycotoxin production capacity of ten <span class="elsevierStyleItalic">A&#46; fumigatus</span> strains isolated from farm and clinical samples&#46; Farm strains were isolated from milk samples taken from dairy cows in C&#243;rdoba province&#44; some of which were associated with subclinical mastitis&#46; A culture medium was defined to optimize fumagillin production and a detection method was developed by HPLC chromatography&#46; It is known that in addition to the host immune status&#44; strain virulence is a fundamental characteristic that will determine its pathogenicity and&#44; in this sense&#44; fumagillin is considered to be among the virulence factors&#46; In the present work&#44; all the strains tested for the production of fumagillin were able to synthesize it&#44; highlighting that the strain <span class="elsevierStyleItalic">A&#46; fumigatus</span> RC2243&#44; from a milk sample from a cow with clinical mastitis&#44; was the most productive&#46; The existence of fumagillin-producing strains represents a potential risk of mycotoxins being transferred to raw milk&#44; constituting a public health risk&#46;</p></span>"
      ]
      "es" => array:2 [
        "titulo" => "Resumen"
        "resumen" => "<span id="abst0015" class="elsevierStyleSection elsevierViewall"><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">En Argentina no existen reportes sobre cepas de <span class="elsevierStyleItalic">Aspergillus fumigatus</span> productoras de fumagilina&#46; En este trabajo se describe el aislamiento y la producci&#243;n de dicha micotoxina por 10 cepas&#44; provenientes del medioambiente rural y aisladas de muestras cl&#237;nicas&#46; Las cepas de origen rural fueron aisladas de vacas lecheras en tambos de la provincia de C&#243;rdoba&#44; y algunas de esas cepas se asociaron a casos de mastitis subcl&#237;nica&#46; Se defini&#243; la composici&#243;n de un medio de cultivo para optimizar la producci&#243;n de fumagilina y se desarroll&#243; un m&#233;todo de cromatograf&#237;a HPLC para su determinaci&#243;n&#46; Es conocido que&#44; adem&#225;s del estado inmunitario del hu&#233;sped&#44; la virulencia de la cepa es una de las caracter&#237;sticas fundamentales que determinan su potencial patog&#233;nico y&#44; en este sentido&#44; la fumagilina es considerada un factor de virulencia&#46; En el presente trabajo todas las cepas estudiadas fueron capaces de sintetizarla y la cepa <span class="elsevierStyleItalic">A&#46; fumigatus</span> RC2243&#44; proveniente de leche de una vaca con mastitis subcl&#237;nica&#44; se destac&#243; como la cepa m&#225;s productora&#46; La existencia de cepas productoras de fumagillina representa un riesgo potencial por el pasaje de dicha micotoxina a la leche&#44; lo cual constituye un problema para la salud p&#250;blica&#46;</p></span>"
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          "en" => "<p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">Chromatogram obtained with the injection of 20<span class="elsevierStyleHsp" style=""></span>&#956;l of a 10<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml solution of fumagillin&#46;</p>"
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                  <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="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Origin&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="center" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Strain&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Fumagillin &#40;&#956;g<span class="elsevierStyleHsp" style=""></span>&#183;<span class="elsevierStyleHsp" style=""></span>ml<span class="elsevierStyleSup">&#8722;1</span>&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " rowspan="7" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Cow milk</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2210&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
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                  \t\t\t\t">9&#46;23&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2213&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">9&#46;23&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2229&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">9&#46;97&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2240&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">5&#46;4&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2243&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">16&#46;79&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2249&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">5&#46;47&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2251&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">4&#46;2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " rowspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Human aspergillosis</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2257&#42;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">8&#46;93&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2261&#42;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
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                  \t\t\t\t">12&#46;74&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">RC2263&#42;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="center" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">7&#46;97&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
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          "en" => "<p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">Fumagillin concentration produced by <span class="elsevierStyleItalic">A&#46; fumigatus</span> strains in Peterson&#8211;Goldstein modified medium&#46;</p>"
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    ]
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Article information

ISSN: 03257541
Original language: English

DOI:10.1016/j.ram.2022.03.003

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2024 September	29	20	49
2024 August	15	13	28
2024 July	18	9	27
2024 June	12	2	14
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