Scientific letter
Cryptosporidium hominis IbA12G3: First report of a rare sub-genotype in Spain
Cryptosporidium hominis IbA12G3: primera descripción de un subgenotipo infrecuente en España
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Köster, Isabel Fuentes, David Carmena" "autores" => array:4 [ 0 => array:3 [ "nombre" => "Rosario" "apellidos" => "Millán" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">a</span>" "identificador" => "aff0005" ] ] ] 1 => array:3 [ "nombre" => "Pamela C." "apellidos" => "Köster" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">b</span>" "identificador" => "aff0010" ] ] ] 2 => array:3 [ "nombre" => "Isabel" "apellidos" => "Fuentes" "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">b</span>" "identificador" => "aff0010" ] ] ] 3 => array:4 [ "nombre" => "David" "apellidos" => "Carmena" "email" => array:1 [ 0 => "dacarmena@isciii.es" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">b</span>" "identificador" => "aff0010" ] 1 => array:2 [ "etiqueta" => "<span class="elsevierStyleSup">*</span>" "identificador" => "cor0005" ] ] ] ] "afiliaciones" => array:2 [ 0 => array:3 [ "entidad" => "Microbiology and Clinical Parasitology Service, University Hospital Puerta de Hierro, Majadahonda, Madrid, Spain" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Parasitology Reference and Research Laboratory, National Centre for Microbiology, Health Institute Carlos III, Madrid, Spain" "etiqueta" => "b" "identificador" => "aff0010" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "titulosAlternativos" => array:1 [ "es" => array:1 [ "titulo" => "<span class="elsevierStyleItalic">Cryptosporidium hominis</span> IbA12G3: primera descripción de un subgenotipo infrecuente en España" ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0005" "etiqueta" => "Fig. 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 2415 "Ancho" => 2520 "Tamanyo" => 324997 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Phylogenetic tree illustrating the evolutionary relationship of the IbA12G3 sequence generated in the present study (represented by a red filled circle) to other <span class="elsevierStyleItalic">C. hominis</span> family Ib sequences previously identified in Spain and other countries at the <span class="elsevierStyleItalic">GP60</span> locus inferred by a neighbour-joining analysis. Red empty circles represent all human (<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>5) and non-human (<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>1) primate IbA12G3 sequences reported globally to date. Black filled circles indicate reference sequences from GenBank<span class="elsevierStyleSup">®</span>. Bootstrapping values over 50% from 1.000 replicates are shown at the branch points. The evolutionary distances were computed using the Tamura 3-parameter method. The rate variation among sites was modelled with a gamma distribution (shape parameter<span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>1). <span class="elsevierStyleItalic">C. parvum</span> was used as outgroup taxa.</p>" ] ] ] "textoCompleto" => "<span class="elsevierStyleSections"><p id="par0005" class="elsevierStylePara elsevierViewall">Members of the genus <span class="elsevierStyleItalic">Cryptosporidium</span> are major contributors to the burden of diarrhoeal disease globally.<a class="elsevierStyleCrossRef" href="#bib0060"><span class="elsevierStyleSup">1</span></a> Cryptosporidiosis primarily affect children in resource-poor settings with unsafe drinking water and inadequate sanitary facilities, but also represents a significant health concern in developed nations.<a class="elsevierStyleCrossRef" href="#bib0065"><span class="elsevierStyleSup">2</span></a><span class="elsevierStyleItalic">Cryptosporidium hominis</span> and <span class="elsevierStyleItalic">Cryptosporidium parvum</span>, particularly the former, are the two major causative species of human cryptosporidiosis. To date, at least nine subtype families (Ia, Ib, Id, Ie, If, Ig, Ih, Ii, Ik) have been identified within <span class="elsevierStyleItalic">C. hominis</span> by sequence analysis of the 60-kDa glycoprotein (<span class="elsevierStyleItalic">gp60</span>) gene.<a class="elsevierStyleCrossRef" href="#bib0070"><span class="elsevierStyleSup">3</span></a> Among them, Ia (1.7%), Ib (92.2%), Id (5.1%), and Ie (0.9%) were the subtype families found (<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>529) in the four largest molecular epidemiological surveys conducted so far in Spain.<a class="elsevierStyleCrossRefs" href="#bib0075"><span class="elsevierStyleSup">4–7</span></a> In these studies, all the Ib samples (<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>152) molecularly characterized at the sub-genotype level were assigned to IbA10G2.</p><p id="par0010" class="elsevierStylePara elsevierViewall">In December 2017 an 18-month-old male infant complaining of gastrointestinal symptoms including altered intestinal transit, distended abdomen, cramps, and acute, non-bloody watery diarrhoea associated to weigh loss (below 25th percentile) and anaemia (serum iron: 24<span class="elsevierStyleHsp" style=""></span>μg/dL) was admitted to the outpatient clinic of the University Hospital Puerta de Hierro Majadahonda (Madrid) for routine coproparasitological examination. The patient had a normal immune status, no contact with pet animals, and no relevant record of recent travelling abroad, although his father reported travelling to Romania during the same period. A single, concentrated stool sample tested positive for the presence of <span class="elsevierStyleItalic">Cryptosporidium</span> oocysts by a rapid immunochomatographic test (Cer Test Biotec S.L., Zaragoza, Spain) and by microscopic examination of a fresh faecal smear stained with the modified Ziehl–Neelsen method. As part of an ongoing research project, a new, fresh aliquot of the faecal material was sent to the National Centre for Microbiology at Majadahonda (Madrid) for genotyping analyses. Total DNA was extracted and purified using the QIAamp<span class="elsevierStyleSup">®</span> DNA stool mini test kit (Qiagen, Hilden, Germany). Molecular characterization of the sample was achieved by PCR amplification of the <span class="elsevierStyleItalic">gp60</span> marker.<a class="elsevierStyleCrossRef" href="#bib0095"><span class="elsevierStyleSup">8</span></a> Amplicons of the expected size (∼870<span class="elsevierStyleHsp" style=""></span>bp) were directly submitted for sequencing. Subsequent sequence analyses confirmed the presence of <span class="elsevierStyleItalic">C. hominis</span> 1bA12G3, a sub-genotype not previously identified in Spain. A representative nucleotide sequence of the sub-genotype obtained was submitted to the GenBank<span class="elsevierStyleSup">®</span> public repository under the accession number MH161561.</p><p id="par0015" class="elsevierStylePara elsevierViewall">Remarkably, a search using the BLAST tool of the National Centre for Biotechnology Information (NCBI) revealed that only six additional IbA12G3 sequences have been previously deposited in GenBank<span class="elsevierStyleSup">®</span>. This particular <span class="elsevierStyleItalic">C. hominis</span> sub-genotype was initially described in a human specimen from UK in 2010.<a class="elsevierStyleCrossRef" href="#bib0100"><span class="elsevierStyleSup">9</span></a> Further molecular research conducted in the Sonora state (Mexico) allowed the identification of IbA12G3 in four children attending hospital settings, three of them presenting with gastrointestinal and/or nutritional disorders and the remaining one asymptomatic at the moment of diagnosis.<a class="elsevierStyleCrossRef" href="#bib0105"><span class="elsevierStyleSup">10</span></a> Finally, IbA12G3 has also been reported in seven rhesus macaques (<span class="elsevierStyleItalic">Macaca mulatta</span>) housed on monkey farms in China, a finding that may be indicative of potential zoonotic transmission.<a class="elsevierStyleCrossRef" href="#bib0110"><span class="elsevierStyleSup">11</span></a><a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a> shows the phylogenetic relationships among the IbA12G3 sequences generated in the present study and those reported in the surveys mentioned above. Appropriated reference sequences and representative sequences of the most frequent <span class="elsevierStyleItalic">C. hominis</span> family subtypes circulating in Spanish human populations were retrieved from NCBI and included in the analysis for comparative purposes. As expected, sequences belonging to the family subtype Ib grouped together in a well-defined cluster. Within this group, the IbA12G3 sequence of non-human primate origin was placed into an independent (but closely related) clade, very likely reflecting differences in host adaptation and specificity.</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0020" class="elsevierStylePara elsevierViewall">In summary, we show here the first description in Spain of a <span class="elsevierStyleItalic">C. hominis</span> human infection by IbA12G3, a very rare sub-genotype only reported before in clinical specimens in UK and Mexico. A host-adapted IbA12G3 variant has been previously identified infecting non-human primates in China, raising doubts about its actual zoonotic potential. Although the origin of the infection in our paediatric case could not be elucidated, we cannot rule out the possibility that this <span class="elsevierStyleItalic">C. hominis</span> sub-genotype is naturally circulating in Spain at low frequency rates. This study clearly demonstrates that molecular surveillance of human cryptosporidiosis is key in determining the occurrence or emergence of novel genotypes of the parasite of uncertain pathogenicity and virulence.</p><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0005">Funding</span><p id="par0025" class="elsevierStylePara elsevierViewall">This study was funded by the <span class="elsevierStyleGrantSponsor" id="gs1">Health Institute Carlos III (ISCIII), Ministry of Economy and Competitiveness</span> under project MPY 1350/16.</p></span></span>" "textoCompletoSecciones" => array:1 [ "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0005" "titulo" => "Funding" ] 1 => array:2 [ "identificador" => "xack399976" "titulo" => "Acknowledgements" ] 2 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "multimedia" => array:1 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Fig. 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 2415 "Ancho" => 2520 "Tamanyo" => 324997 ] ] "descripcion" => array:1 [ "en" => "<p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Phylogenetic tree illustrating the evolutionary relationship of the IbA12G3 sequence generated in the present study (represented by a red filled circle) to other <span class="elsevierStyleItalic">C. hominis</span> family Ib sequences previously identified in Spain and other countries at the <span class="elsevierStyleItalic">GP60</span> locus inferred by a neighbour-joining analysis. Red empty circles represent all human (<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>5) and non-human (<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>1) primate IbA12G3 sequences reported globally to date. Black filled circles indicate reference sequences from GenBank<span class="elsevierStyleSup">®</span>. Bootstrapping values over 50% from 1.000 replicates are shown at the branch points. The evolutionary distances were computed using the Tamura 3-parameter method. The rate variation among sites was modelled with a gamma distribution (shape parameter<span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>1). <span class="elsevierStyleItalic">C. parvum</span> was used as outgroup taxa.</p>" ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0015" "bibliografiaReferencia" => array:11 [ 0 => array:3 [ "identificador" => "bib0060" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for <span class="elsevierStyleItalic">Cryptosporidium</span>" "autores" => array:1 [ 0 => array:2 [ "etal" => true "autores" => array:6 [ 0 => "W. Checkley" 1 => "A.C. White" 2 => "D. Jaganath" 3 => "M.J. Arrowood" 4 => "R.M. Chalmers" 5 => "X.M. 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| Year/Month | Html | Total | |
|---|---|---|---|
| 2024 November | 6 | 1 | 7 |
| 2024 October | 14 | 21 | 35 |
| 2024 September | 31 | 16 | 47 |
| 2024 August | 23 | 11 | 34 |
| 2024 July | 22 | 10 | 32 |
| 2024 June | 18 | 8 | 26 |
| 2024 May | 22 | 6 | 28 |
| 2024 April | 16 | 27 | 43 |
| 2024 March | 31 | 20 | 51 |
| 2024 February | 30 | 11 | 41 |
| 2024 January | 40 | 20 | 60 |
| 2023 December | 29 | 20 | 49 |
| 2023 November | 27 | 17 | 44 |
| 2023 October | 29 | 43 | 72 |
| 2023 September | 20 | 27 | 47 |
| 2023 August | 24 | 10 | 34 |
| 2023 July | 19 | 105 | 124 |
| 2023 June | 25 | 27 | 52 |
| 2023 May | 56 | 3 | 59 |
| 2023 April | 35 | 4 | 39 |
| 2023 March | 19 | 4 | 23 |
| 2023 February | 17 | 6 | 23 |
| 2023 January | 12 | 14 | 26 |
| 2022 December | 26 | 10 | 36 |
| 2022 November | 33 | 25 | 58 |
| 2022 October | 24 | 22 | 46 |
| 2022 September | 29 | 22 | 51 |
| 2022 August | 38 | 22 | 60 |
| 2022 July | 32 | 14 | 46 |
| 2022 June | 35 | 14 | 49 |
| 2022 May | 28 | 33 | 61 |
| 2022 April | 53 | 30 | 83 |
| 2022 March | 69 | 23 | 92 |
| 2022 February | 107 | 10 | 117 |
| 2022 January | 59 | 13 | 72 |
| 2021 December | 41 | 32 | 73 |
| 2021 November | 52 | 26 | 78 |
| 2021 October | 48 | 18 | 66 |
| 2021 September | 28 | 17 | 45 |
| 2021 August | 25 | 9 | 34 |
| 2021 July | 31 | 18 | 49 |
| 2021 June | 21 | 9 | 30 |
| 2021 May | 28 | 12 | 40 |
| 2021 April | 35 | 8 | 43 |
| 2021 March | 43 | 9 | 52 |
| 2021 February | 24 | 7 | 31 |
| 2021 January | 17 | 13 | 30 |
| 2020 December | 19 | 6 | 25 |
| 2020 November | 21 | 16 | 37 |
| 2020 October | 19 | 5 | 24 |
| 2020 September | 42 | 14 | 56 |
| 2020 August | 28 | 21 | 49 |
| 2020 July | 36 | 15 | 51 |
| 2020 June | 29 | 20 | 49 |
| 2020 May | 24 | 17 | 41 |
| 2020 April | 16 | 10 | 26 |
| 2020 March | 23 | 8 | 31 |
| 2020 February | 36 | 7 | 43 |
| 2020 January | 32 | 7 | 39 |
| 2019 December | 37 | 20 | 57 |
| 2019 November | 18 | 10 | 28 |
| 2019 October | 24 | 16 | 40 |
| 2019 September | 3 | 6 | 9 |
| 2019 August | 0 | 10 | 10 |
| 2019 July | 2 | 6 | 8 |
| 2019 June | 3 | 11 | 14 |
| 2019 May | 2 | 69 | 71 |
| 2019 April | 22 | 21 | 43 |
| 2019 March | 25 | 8 | 33 |
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