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. 2025 Jun 8;15(12):1695.
doi: 10.3390/ani15121695.

Microsporidia in Rodents- Mus musculus, Rattus norvegicus, and Rattus rattus-A Public Health Concern in the Canary Islands, Spain

Sergio Llorens-Berzosa  1 , Edgar Baz-González  2   3   4 , Natalia Martin-Carrillo  2   3 , Katherine García-Livia  2   3 , Virginia Amaro-Ramos  2   3 , Néstor Abreu-Acosta  3   5 , Carmen Del Aguila  1 , Jordi Miquel  6   7 , Román Pino-Vera  2   3   4 , Estefanía Abreu-Yanes  5 , Carlos Feliu  6 , Fernando Izquierdo  1 , Pilar Foronda  2   3
Affiliations

Microsporidia in Rodents- Mus musculus, Rattus norvegicus, and Rattus rattus-A Public Health Concern in the Canary Islands, Spain

Sergio Llorens-Berzosa et al. Animals (Basel). .

Abstract

Rodents are recognized as reservoirs of a wide range of pathogens, including microsporidia. The presence of microsporidia in the environment of mainland Spain and its islands has become increasingly known, as the number of studies has multiplied over time. The present study was conducted to determine the occurrence and diversity of microsporidia in three rodent species (Mus musculus, Rattus norvegicus, and Rattus rattus) in the Canary Islands, Spain. Ninety-three fecal samples were obtained from wild rodents on La Gomera and Gran Canaria Islands. Each sample was tested using Weber's modified trichrome staining and immunofluorescence antibody tests (IFATs) against the Encephalitozoon genus and Enterocytozoon bieneusi. The microscopy-positive samples were subsequently analyzed using a nested polymerase chain reaction (PCR) followed by Sanger sequencing. The staining technique showed 38.7% (36/93) positivity, whereas the IFATs for Encephalitozoon spp. and Ent. bieneusi revealed 3.2% (3/93) and 6.5% (6/93) positivity, respectively. Finally, the nested PCR and nucleotide sequence analysis confirmed a 9.7% (9/93) occurrence of Ent. bieneusi and 17.2% occurrence (16/93) of different undetermined microsporidia species, whereas no Encephalitozoon spp. were detected. Seven different Ent. bieneusi genotypes were detected as follows: three known (AAE1, D, and SBM1) and four novel (GRE1, GRE2, LGE1, and LGE2), all of which belonged to Group 1. The results demonstrate, for the first time, that microsporidia are present in the rodent populations of the Canary Islands. Further studies are needed to determine the impact of the presence of microsporidia in rodents on the zoonotic transmission of these parasites.

Keywords: Canary Islands; Encephalitozoon spp.; Enterocytozoon bieneusi; microsporidia; rodents; zoonotic potential.

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Conflict of interest statement

Authors Néstor Abreu-Acosta and Estefanía Abreu-Yanes were employed by the company Nertalab S.L. company. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Map of the sampled areas on the islands of La Gomera and Gran Canaria (Canary Islands, Spain). The upper-right corner shows the Canary Islands, as they are related to Africa and mainland Spain. The upper-left panel shows the Canarian archipelago. The sampled municipalities of La Gomera Island are as follows: 1. Vallehermoso, 2. Agulo, 3. Hermigua, 4. San Sebastián de La Gomera, 5. Alajeró, and 6. Valle Gran Rey. Those of Gran Canaria Island are as follows: 7. Artenara, 8. Firgas, 9. Arucas, and 10. Ingenio. Adapted from Wikimedia Common (https://upload.wikimedia.org/wikipedia/commons/d/d1/Mapa_Canarias_municipios.svg, accessed on 10 March 2025; https://upload.wikimedia.org/wikipedia/commons/2/28/Islas_Canarias_%28real_location%29_in_Spain.svg, accessed on 10 March 2025), License CC BY-SA 3.0 (https://creativecommons.org/licenses/by-sa/3.0/, accessed on 26 May 2025), by which permission to copy, distributed, or adapt was established. Users: TUBS (https://commons.wikimedia.org/wiki/User:TUBS, accessed on 10 March 2025), Tintazul (https://commons.wikimedia.org/wiki/User:Tintazul, accessed on 10 March 2025). GRAFCAN and IDE Canarias (Source: Gobierno de Canarias (https://www3.gobiernodecanarias.org/medusa/mediateca/ecoescuela/?attachment_id=3322, accessed on 26 May 2025; https://www3.gobiernodecanarias.org/medusa/mediateca/ecoescuela/?attachment_id=3265, accessed on 26 May 2025), License CC BY-NC-ND 4.0 (https://creativecommons.org/licenses/by-nc-nd/4.0/, accessed on 26 May 2025), images modified with permission of the licensor.
Figure 2
Figure 2
(A). Weber’s chromotrope stain of the intestinal contests of Rattus norvegicus showing three spore-compatible structures clustered together (B). Mus musculus showing a single spore-compatible structure (100×) (C). Encephalitozoon spp. spore-compatible structures were visualized by IFAT using mAb 2C2 (100×) in a sample from Rattus rattus (D). Cluster of spore-compatible structures with Enterocytozoon bieneusi visualized by IFAT using mAb 6E52D9 (100×) in a sample from M. musculus.
Figure 3
Figure 3
The phylogenetic relationships between the sequence of the internal transcribed spacer (ITS) region of the ribosomal RNA gene of the Enterocytozoon bieneusi genotypes obtained in this study and the sequences of known genotypes deposited in GenBank. The Neighbor-Joining method was used based on the genetic distance calculated using the Kimura 2-parameter model. Reference sequences from each phylogenetic group of Ent. bieneusi genotypes (Groups 1–15) were used. The accession numbers are shown in bold, and information regarding the host species and origin is shown in parentheses. There were a total of 240 positions in the final dataset.
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