Bibliographic Details
| Title: |
The production of Necator americanus larvae for use in experimental human infection |
| Authors: |
Paul R. Chapman, Stacey Llewellyn, Helen Jennings, Luke Becker, Paul Giacomin, Rodney McDougall, Jennifer Robson, Alex Loukas, James McCarthy |
| Source: |
Parasites & Vectors, Vol 15, Iss 1, Pp 1-7 (2022) |
| Publisher Information: |
BMC, 2022. |
| Publication Year: |
2022 |
| Collection: |
LCC:Infectious and parasitic diseases |
| Subject Terms: |
Hookworm culture, Larval larvae, Hookworm larvae production, Infectious and parasitic diseases, RC109-216 |
| More Details: |
Abstract Background Although there is unprecedented interest in experimental human hookworm infection, details of hookworm manufacture and characterisation have been sparsely reported. In this report, we detail the production and characterisation of Necator americanus larvae for use in a recently published clinical trial. Methods Faeces was obtained from an experimentally infected donor. Faecal hookworm DNA was determined by quantitative PCR. Paired samples were incubated in either sterile water or sterile water mixed with antimicrobials (amphotericin and gentamicin). Coproculture was performed by modified Harada-Mori method. The harvested larvae were then processed in either sterile water or antiseptic solution. Larval yield was then calculated (larvae per gram), larval viability was determined by thermally induced motility assay and microbial burden was determined at the day of harvest, at 48 h and at 7 days. Results Twenty-eight faecal cultures were performed over 16 months. The faecal hookworm DNA content was variable over this time. There was no association of larval yield with faecal hookworm DNA content. Pre-treatment of faeces with antimicrobials did not influence larval yield. Larval motility was 85.3% (95% CI 79.3–91.3%). Incubation of larvae in antiseptics did not reduce viability at 14 days with a marginal mean of 68.6% (95% CI 59.1–78.1%) washed in water vs. 63.3% (95% CI 53.8 – 72.9%) when incubated in betadine (p = 0.38). Larvae washed in sterile water did not meet microbial bioburden criteria. Incubation in antiseptic resulted in acceptable microbial bioburden at 48 h but not at 7 days. Although the addition of gentamicin did reduce the microbial bio-burden acceptable levels, it was found to significantly lower larval motility at 7 days compared to incubation in sterile water and motility at 7 days 37.8% (95% CI 4.7–70.9%) vs. 67.3% (95% CI 35.2–99.3%, p |
| Document Type: |
article |
| File Description: |
electronic resource |
| Language: |
English |
| ISSN: |
1756-3305 |
| Relation: |
https://doaj.org/toc/1756-3305 |
| DOI: |
10.1186/s13071-022-05371-y |
| Access URL: |
https://doaj.org/article/c69223f0b6ec4c078a1f258c3175e896 |
| Accession Number: |
edsdoj.69223f0b6ec4c078a1f258c3175e896 |
| Database: |
Directory of Open Access Journals |
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