Authors

E Ukaegbu⁵; U Anjum¹; S Llorens²; L Acosta³; G Torrado⁴; MD Evans⁵; A Peña-Fernández⁴;  ¹ Leicester School of Allied Health Sciences, De Montfort University, Sierra Leone;  ² Facultad de Farmacia, Universidad San Pablo CEU, Spain;  ³ Área de Parasitología, Departamento de Agroquímica y Medio Ambiente, Universidad Miguel Hernández de Elche, Spain;  ⁴ Departamento de Ciencias Biomédicas, Universidad de Alcalá, Crta, Spain;  ⁵ Department of Public Health, University of Makeni, Sierra Leone

Discussion

Following the detection of emerging microsporidian spores in faeces from different farm animals collected in April 2019 from different farms across Bombali District, Sierra Leone (SL), De Montfort University (DMU, UK) started the implementation of a multistage project to build medical parasitology capabilities at the University of Makeni (UniMak, SL) to tackle potential future risks due to these opportunistic parasites.
Aims: a) to explore the temporal variation of human-related microsporidian parasites in different farms’ pigs across Makeni to identify potential risks for the food chain; b) to analyse the effectiveness of our interventions/stages to provide UniMak BSc Public Health students with the appropriate laboratory skills to perform coprological analysis for the detection of these species.
UniMak academics introduced novel practicals for the detection of microsporidia in third- and four-year BSc Public Health’s modules in 2020/21. Academics followed a blended approach using the Virtual Laboratory freely available in the e-Parasitology package (http://parasitology.dmu.ac.uk/). Students processed thirty-six fresh stool samples from pigs provided by the SL Department of Agriculture and Food Security from a nearby farm (Makeni) in 2021. Students prepared smears in a class II biological safety cabinet and learned to perform modified Trichrome stain for the detection of spores. Moreover, in order to explore temporal variation, further fresh faecal samples were collected from four pigs in Summer 2022. These recent samples were specifically screened for the presence of Encephalitozoon spores by immunofluorescent-antibody test using a specific monoclonal antibody (Mab) of murine origin IgG2a, patented as a diagnostic tool for miscrosporidiasis caused by the genus Encephalitozoon.
Students showed high levels of confidence as soon as they entered the laboratory. They were able to quickly undertake all the different practical steps. A high level of accuracy was observed in students’ detection of spores, which were present in 14/36 (38.8%) stool smears monitored and would be in line with the prevalence of microsporidia detected previously (7/12; 58%) by our group using a SYBR Green real-time PCR on 12 pig stool samples collected in 2019. One of the samples collected in 2022 resulted positive for spores of the genus Encephalitozoon. Moreover, structures compatible with Encephalitozoon spp. were observed in faeces from another pig. These results would be similar to those observed in 2019, suggesting a moderate circulation of Encephalitozoon spp. in the monitored farms, which highlights a potential risk to the Sierra Leonean population, especially those with the immune system compromised, including survivors from the 2014-16 Ebola outbreak.
The different stages and strategies used seem to have successfully aided capacity building to prepare future UniMak professionals to tackle microsporidian infections in Sierra Leone, methods and resour