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Fungal peritonitis by Thermoascus crustaceus in a peritoneal dialysis patient from Chile
Peritonitis fúngica por Thermoascus crustaceus en un paciente en diálisis peritoneal de Chile
Eduardo Alvareza,
Corresponding author
ealvarezd@med.uchile.cl

Corresponding author.
, Alvaro Castillob,c, Isabel Iturrietab,d
a Mycology Unit/Filamentous Fungi – Biomedical Sciences Institute – Microbiology and Mycology Program – Universidad de Chile, Chile
b Hospital San Juan de Dios, La Serena, Chile
c Universidad Católica del Norte, La Serena, Chile
d Universidad Pedro de Valdivia, La Serena, Chile
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    "textoCompleto" => "<span class="elsevierStyleSections"><p id="par0005" class="elsevierStylePara elsevierViewall">An 83-year-old female patient&#44; with a history of cholecystectomy&#44; hernia repair&#44; and hip and knee osteoarthritis was admitted to our hospital&#46; In 1990 a parathyroidectomy was performed&#44; evolving with postoperative hypoparathyroidism&#44; which was treated with calcitriol and oral calcium&#46; Moreover&#44; the patient had a history of severe arterial hypertension since 1992&#44; managed with diuretics&#44; calcium channel blockers and angiotensin-converting enzyme inhibitors&#46; In 1995 the patient developed chronic renal failure&#46; After 10 years she progressed to chronic kidney disease stage 5&#44; opting for peritoneal dialysis&#46; A Tenkhoff catheter was put in 2002&#46; During the first 2 years of peritoneal dialysis the patient developed two episodes of Gram-positive bacterial peritonitis&#44; both completely cured&#46;</p><p id="par0010" class="elsevierStylePara elsevierViewall">In 2012&#44; a permanent pacemaker was implanted due to symptomatic congenital atrioventricular block&#46; In addition&#44; a new bacterial peritonitis due to <span class="elsevierStyleItalic">Corynebacterium</span> spp&#46; was observed&#46; The patient was successfully treated&#46;</p><p id="par0015" class="elsevierStylePara elsevierViewall">On January 2014 the patient presented with pain and cloudy peritoneal fluid&#46; The study of the dialysate fluid showed 80 white blood cells&#47;mm<span class="elsevierStyleSup">3</span>&#46; No differential count was performed&#46; Bacterial cultures were negative after 7 days of incubation at 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The patient received routine intraperitoneal antibiotics&#46;</p><p id="par0020" class="elsevierStylePara elsevierViewall">In June 2014&#44; after a mild story of two months of intermittent fever peaking mainly in the evening&#44; with occasional cloudy effluent outlet&#44; new microbiological studies were performed&#46; Clinically&#44; the patient presented no abdominal pain&#46; Again&#44; bacterial cultures were negative after 7 days of incubation&#46; Gram staining did not show any microbiological evidence&#46; However&#44; dialysate fluid cytology showed 260 white blood cells&#47;mm<span class="elsevierStyleSup">3</span> &#40;68&#37; mononuclear cells&#41;&#46; In addition&#44; blood culture bottles inoculated with peritoneal fluid&#44; indicated the presence of fungal growth&#46; Growth on Sabouraud Dextrose Agar &#40;SDA&#41; showed the presence of yellow-light brown colonies&#46; <a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a> shows the radiographic findings&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0025" class="elsevierStylePara elsevierViewall">Antifungal treatment was started with intraperitoneal amphotericin B 50<span class="elsevierStyleHsp" style=""></span>mg&#47;day for 20 days&#46; The patient completed 10 days of treatment after which the indwelling Tenckhoff catheter was removed&#46; Following antifungal treatment&#44; the patient was discharged in good condition&#46;</p><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Mycological studies</span><p id="par0030" class="elsevierStylePara elsevierViewall">The isolate was referred to the Fungal Reference Centre at Universidad de Chile&#46; The mould was cultured on Potato Dextrose Agar &#40;PDA&#41; and Corn Meal Agar &#40;CMA&#41; for 5 days at 25<span class="elsevierStyleHsp" style=""></span>&#176;C and 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The growth of the isolate on PDA &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>A&#41; and CMA revealed colonies that were initially flat and buff coloured but that quickly &#40;within 6 days&#41; became yellowish orange with a brownish yellow reverse&#46; Sparse conidial structures of the anamorph <span class="elsevierStyleItalic">Paecilomyces</span> were evident in 3&#8211;4 days at 25<span class="elsevierStyleHsp" style=""></span>&#176;C on PDA and CMA&#46; Branched conidiophores measuring between 350 and 400<span class="elsevierStyleHsp" style=""></span>&#956;m were observed&#46; Conidia were elliptical to subglobose&#44; approximately 7&#8211;7&#46;8 by 4&#46;5&#8211;5&#46;3<span class="elsevierStyleHsp" style=""></span>&#956;m&#44; smooth-walled in long chains &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>B and C&#41;&#46; In order to induce the formation of sexual stage&#44; additional subcultures onto malt extract agar &#40;MEA&#41; &#40;Becton Dickinson&#41; at 37<span class="elsevierStyleHsp" style=""></span>&#176;C were performed&#46; Colonies on MEA became brown-orange and granular to crust-like with a brownish orange reverse&#46; Subspherical&#44; nonostiolate ascomata occurred within 7 days and contained asci that were approximately 600 by 750<span class="elsevierStyleHsp" style=""></span>&#956;m &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>D&#41;&#46; Ascospores were elliptical&#44; pale yellow&#44; thick walled&#44; approximately 4&#8211;5 by 6&#46;5&#8211;7<span class="elsevierStyleHsp" style=""></span>&#956;m&#44; and predominately echinulate by light microscopy &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>E&#41;&#46; Temperature studies were also performed by inoculating five plates of MEA in duplicate with a 1<span class="elsevierStyleHsp" style=""></span>mm portion of the isolate&#46; Plates were incubated at 15&#44; 25&#44; 37&#44; and 42<span class="elsevierStyleHsp" style=""></span>&#176;C for 6 days&#44; and zone sizes were measured&#46; Optimal growth was observed at 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; The ascocarp formation occurred mainly at 37<span class="elsevierStyleHsp" style=""></span>&#176;C &#40;<a class="elsevierStyleCrossRef" href="#fig0010">Fig&#46; 2</a>F&#41;&#46;</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia><p id="par0035" class="elsevierStylePara elsevierViewall">In addition&#44; an in order to corroborate the morphological identification&#44; molecular studies based on sequences of the ITS region were conducted&#46; PCR amplification of fungal genomic DNA was performed using the primers and conditions as described previously&#46;<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">1</span></a> The results obtained from GENBANK showed a high level of identity &#40;&#8805;99&#37;&#41; with the sequences &#40;accession number <a href="ncbi-n:FN675837">FN675837</a>&#41; from the type strain of <span class="elsevierStyleItalic">Thermoascus crustaceus</span> &#40;<span class="elsevierStyleItalic">T&#46; crustaceus</span> CBS 181&#46;67<span class="elsevierStyleSup">T</span>&#41;&#46; In order to evaluate the genetical relationships between our isolate and those reported previously&#44; phylogenetical analyses were conducted &#40;<a class="elsevierStyleCrossRef" href="#fig0015">Fig&#46; 3</a>&#41;&#46; Phylogenetical inference was carried out following the protocols reported by Alvarez et al&#46;<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">2</span></a> The isolate reported in the present study has been stored in the Chilean Fungal Collection &#40;ChFC&#41; housed at the Mycology Unit&#44; University of Chile&#44; Santiago de Chile&#44; with the unique identifier ChFC EA144&#46; Also&#44; the sequence of ITS region from our isolate was deposited in GENBANK database as KY115690&#46;</p><elsevierMultimedia ident="fig0015"></elsevierMultimedia></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Susceptibility studies</span><p id="par0040" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">In vitro</span> susceptibility testing was performed using broth microdilution for filamentous fungi according to the CLSI document M38-A2&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">5</span></a> The MIC and MEC &#40;minimal effective concentration&#41; values were read at 48 and 72<span class="elsevierStyleHsp" style=""></span>h&#46; Due to the lack of clinical breakpoints for <span class="elsevierStyleItalic">Thermoascus</span> species&#44; the following suggested cut off breakpoints were used&#58; sensible &#40;S&#41;&#44; &#8804;1<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#59; intermediate &#40;I&#41;&#44; 2<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#59; and resistant &#40;R&#41;&#44; &#8805;4<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#46;<a class="elsevierStyleCrossRef" href="#bib0085"><span class="elsevierStyleSup">5</span></a> Pure active powders of known potency of amphotericin B &#40;AMB&#41; &#40;USP&#44; Rockville&#44; MD&#41;&#44; voriconazole &#40;VRC&#41; &#40;Sigma-Aldrich&#41;&#44; itraconazole &#40;ITC&#41; &#40;Janssen Pharmaceutica&#44; Beerse&#44; Belgium&#41;&#44; caspofungin &#40;CSF&#41; &#40;Merk &#38; Co&#46;&#44; Inc&#46;&#44; Rahway&#44; USA&#41;&#44; micafungin &#40;MCF&#41; &#40;Astellas Pharma&#41;&#44; and anidulafungin &#40;ANF&#41; &#40;Pfizer&#41; were tested&#46; All tests were performed in duplicates&#46; <span class="elsevierStyleItalic">Paecilomyces variotii</span> ATCC MYA 3630 was included as quality control&#46;</p><p id="par0045" class="elsevierStylePara elsevierViewall">Results indicated the good activity of the antifungals tested&#46; Amphotericin B and azoles showed MIC values of 0&#46;5 and 0&#46;25<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#44; respectively&#46; Equinocandins showed the best activity with MIC values of 0&#46;125<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#46; Based upon achievable drug concentrations using standard dosing regimens&#44; the isolate appeared to be sensible&#44; <span class="elsevierStyleItalic">in vitro</span>&#44; to all the antifungal agents assayed in the present work&#46;</p></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Discussion</span><p id="par0050" class="elsevierStylePara elsevierViewall">Fungal peritonitis is a rare complication in patients undergoing peritoneal dialysis&#46; The incidence is around 6&#37;&#46;<a class="elsevierStyleCrossRefs" href="#bib0080"><span class="elsevierStyleSup">4&#44;12</span></a> Etiological agents reported to date include <span class="elsevierStyleItalic">Aspergillus</span> spp&#46;&#44;<a class="elsevierStyleCrossRef" href="#bib0075"><span class="elsevierStyleSup">3</span></a><span class="elsevierStyleItalic">Candida</span> spp&#46;&#44;<a class="elsevierStyleCrossRefs" href="#bib0080"><span class="elsevierStyleSup">4&#44;12</span></a><span class="elsevierStyleItalic">Curvularia geniculata</span>&#44;<a class="elsevierStyleCrossRef" href="#bib0115"><span class="elsevierStyleSup">11</span></a><span class="elsevierStyleItalic">Fusarium</span> spp&#46;&#44;<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">7</span></a><span class="elsevierStyleItalic">Penicillium</span> spp&#46;&#44;<a class="elsevierStyleCrossRef" href="#bib0105"><span class="elsevierStyleSup">9</span></a> and <span class="elsevierStyleItalic">T&#46; crustaceus</span>&#44;<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">8</span></a> among others&#46; Recently&#44; following the Chile&#39;s earthquake of 2010&#44; several cases of fungal peritonitis due to <span class="elsevierStyleItalic">Paecilomyces variotti</span> were reported&#46;<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">10</span></a> A search of the literature indicates that the present case is the second report describing fungal peritonitis by <span class="elsevierStyleItalic">T&#46; crustaceus</span> in the world&#44;<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">8</span></a> and the first in Chile&#46;</p><p id="par0055" class="elsevierStylePara elsevierViewall">In an earlier examination of the fungal cultures&#44; and based on taxonomic information from fungal structures&#44; <span class="elsevierStyleItalic">Paecilomyces</span> species was identified in the plates incubated at room temperature&#46; Teleomorph &#40;<span class="elsevierStyleItalic">Thermoascus</span>&#41; morphological structures were observed from agar plates cultured at 37<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; It was also possible to confirm the preliminary morphological identification with the molecular tools&#46; The ITS sequence of the studied isolate allowed us to identify it as <span class="elsevierStyleItalic">T&#46; crustaceus</span>&#46;</p><p id="par0060" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">In vitro</span> susceptibility data for <span class="elsevierStyleItalic">T&#46; crustaceus</span> is limited&#46; However&#44; MIC values for azoles &#40;&#60;0&#46;015<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#41; and amphotericin B &#40;&#60;0&#46;25<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#41; on <span class="elsevierStyleItalic">Thermoascus taitungiacus</span> have been reported&#44;<a class="elsevierStyleCrossRef" href="#bib0090"><span class="elsevierStyleSup">6</span></a> consistent with this study&#46;</p><p id="par0065" class="elsevierStylePara elsevierViewall">Usually fungal peritonitis is a serious disease&#46; In our case the clinical picture was mild&#44; without systemic compromise&#46; Despite the lack of clear treatment guidelines in fungal peritonitis&#44; the early removal of Tenckhoff catheter seems to be essential and timely antifungal treatment must be started in order to have a good prognosis&#46; In the present study&#44; AMB&#44; ITC and CSF&#44; MCF&#44; and ANF were active against the strain of <span class="elsevierStyleItalic">T&#46; crustaceus</span> at low MIC values&#46;</p><p id="par0070" class="elsevierStylePara elsevierViewall">In summary&#44; this is the first report of fungal peritonitis caused by <span class="elsevierStyleItalic">T&#46; crustaceus</span> in Chile&#46; Further studies are needed to elucidate optimal treatment strategies&#46;</p></span></span>"
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      "en" => array:3 [
        "titulo" => "Abstract"
        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Fungal peritonitis is a relatively uncommon infection in peritoneal dialysis patients&#46; However&#44; it can be associated with significant morbimortality&#46; In recent reports&#44; <span class="elsevierStyleItalic">Candida</span> species and other filamentous fungi have been reported as being aetiological agents&#46; <span class="elsevierStyleItalic">Thermoascus</span> species are ubiquitous&#44; thermophilic fungi&#44; with an anamorph in the <span class="elsevierStyleItalic">Paecilomyces</span> genus&#46; Here we present the first report of fungal peritonitis by <span class="elsevierStyleItalic">Thermoascus crustaceus</span> from Chile&#46;</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Case report</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">We present the case of an 83-year-old female patient&#44; with a history of cholecystectomy&#44; hernia repair&#44; severe arterial hypertension&#44; hip and knee osteoarthritis and several episodes of peritoneal dialysis with a cloudy exudate&#46; Bacterial cultures were negative&#46; In addition&#44; a history of two months with intermittent fever peaks mainly in the evening was reported&#46; Blood culture bottles inoculated with peritoneal fluid revealed the presence of fungal growth&#46; Morphological and molecular studies allowed us to identify the aetiological agent as <span class="elsevierStyleItalic">Thermoascus crustaceus</span>&#46; An antifungal susceptibility test was performed using the M38-A2 method&#44; developed by the Clinical and Laboratory Standards Institute &#40;CLSI&#41;&#46; The MIC values to amphotericin B&#44; itraconazole&#44; voriconazole and echinochandins were 0&#46;5&#44; 0&#46;25&#44; 0&#46;25 and 0&#46;125<span class="elsevierStyleHsp" style=""></span>&#956;g&#47;ml&#44; respectively&#46; Antifungal treatment with amphotericin B was prescribed&#44; with good patient progress&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Conclusions</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Fungal peritonitis is a very rare entity&#46; Moreover&#44; the spectrum of fungal pathogens continues to expand&#44; a reason for which morphological and molecular studies are necessary for a rapid diagnosis&#46;</p></span>"
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          0 => array:2 [
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            "titulo" => "Background"
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          1 => array:2 [
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          2 => array:2 [
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        "resumen" => "<span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0030">Antecedentes</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">La peritonitis f&#250;ngica es una infecci&#243;n bastante infrecuente en pacientes en di&#225;lisis peritoneal&#46; Sin embargo&#44; puede estar relacionada con una morbimortalidad considerable&#46; En informes recientes&#44; se han registrado las especies de <span class="elsevierStyleItalic">Candida</span> y algunos hongos filamentosos como agentes etiol&#243;gicos&#46; <span class="elsevierStyleItalic">Thermoascus</span> es un g&#233;nero de hongos ubicuos&#44; term&#243;filos&#44; que tienen su anamorfo en el g&#233;nero <span class="elsevierStyleItalic">Paecilomyces</span>&#46; A continuaci&#243;n presentamos el primer caso de peritonitis f&#250;ngica por <span class="elsevierStyleItalic">Thermoascus crustaceous</span> de Chile&#46;</p></span> <span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Caso cl&#237;nico</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">Presentamos el caso de una mujer de 83 a&#241;os con antecedentes de colecistectom&#237;a&#44; hernia&#44; hipertensi&#243;n arterial grave&#44; artrosis de cadera y rodilla&#44; y varios episodios de di&#225;lisis peritoneal con una efusi&#243;n turbia&#46; Los cultivos bacterianos fueron negativos&#46; Adem&#225;s&#44; la paciente hab&#237;a presentado durante los dos &#250;ltimos meses picos febriles intermitentes&#44; principalmente por la noche&#46; Los hemocultivos inoculados con l&#237;quido peritoneal revelaron el crecimiento de un micelio&#46; Los estudios morfol&#243;gicos y moleculares permitieron identificar el agente etiol&#243;gico como <span class="elsevierStyleItalic">Thermoascus crustaceous</span>&#46; Para el estudio de sensibilidad antif&#250;ngica se utiliz&#243; el m&#233;todo M38-A2&#44; desarrollado por el Clinical and Laboratory Standards Institute&#46; Las CMI obtenidas de anfotericina B&#44; itraconazol&#44; voriconazol y equinocandinas fueron 0&#44;5&#59; 0&#44;25&#59; 0&#44;25&#44; y 0&#44;125&#160;&#956;g&#47;ml&#44; respectivamente&#46; El tratamiento antif&#250;ngico con anfotericina B fue el indicado&#44; con una buena evoluci&#243;n de la paciente&#46;</p></span> <span id="abst0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Conclusiones</span><p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">La peritonitis f&#250;ngica es una entidad muy poco frecuente&#46; Adem&#225;s&#44; el espectro de hongos pat&#243;genos contin&#250;a en expansi&#243;n&#44; raz&#243;n por la cual los estudios morfol&#243;gicos y moleculares son necesarios para el diagn&#243;stico r&#225;pido&#46;</p></span>"
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        0 => array:2 [
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                        0 => array:2 [
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                            5 => "M&#46;G&#46; Rinaldi"
                          ]
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                            1 => "D&#46; Garcia-Hermoso"
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                          "etal" => false
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                            1 => "E&#46;D&#46; Everette"
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Article information
ISSN: 11301406
Original language: English
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