Eustace, A. | bioRxiv Science

bioRxiv2024

Resolving a neonatal intensive care unit outbreak of methicillin-resistant Staphylococcus aureus to the SNV level using Oxford Nanopore simplex reads and HERRO error correction

ObjectivesOur laboratory began prospective genomic surveillance for healthcare-associated organisms in 2022 using Oxford Nanopore Technologies (ONT) sequencing as a standalone platform. This has permitted the early detection of outbreaks but has been insufficient for single-nucleotide variant (SNV)-level analysis due to lower read accuracy than Illumina sequencing. This study aimed to determine whether Haplotype-aware ERRor cOrrection (HERRO) of ONT data could permit high-resolution comparison of outbreak isolates. MethodsWe used ONT simplex reads from isolates involved in a recent outbreak of methicillin-resistant Staphylococcus aureus (MRSA) in our neonatal unit. The raw sequence data were re-basecalled and adapter-trimmed using Dorado v0.7.0. The simplex reads then underwent HERRO correction. The resulting genome assemblies and phylogenies were compared with previous analyses (using Dorado v0.3.4, no HERRO correction and data generated by Illumina sequencing). ResultsFive of nine outbreak isolates were included in the analysis. The remaining four isolates had insufficient read lengths (N50 values <10,000 bp) and did not provide complete chromosome coverage after HERRO correction. The average chromosome sequencing depth for nanopore data was 147x (range: 44-220x) with an average read N50 of 12,215 bp (interquartile range (IQR): 11,439-12,711 bp). The median pairwise SNV distance between outbreak isolates from the original investigation was 51 SNVs (range: 40-68), which decreased to 3 SNVs (range: 1-15) with HERRO correction. Illumina sequencing generated a median SNV distance of 2 (range: 0-13). The resulting standalone ONT HERRO-corrected phylogeny was almost indistinguishable from the standalone Illumina-generated phylogeny. ConclusionsThe addition of HERRO correction meant isolates from this MRSA outbreak could be resolved to a level on par with Illumina sequencing. ONT data following HERRO correction represents a viable standalone option for high-resolution genomic analysis of hospital outbreaks, provided sufficient read lengths can be generated.

genomics↗

bioRxiv2020

Protected areas network is not adequate to protect a critically endangered East Africa Chelonian: Modelling distribution of pancake tortoise, Malacochersus tornieri under current and future climates

While the international pet trade and habitat destruction have been extensively discussed as major threats to the survival of the pancake tortoise (Malacochersus tornieri), the impact of climate change on the species remains unknown. In this study, we used species distribution modelling to predict the current and future distribution of pancake tortoises in Zambezian and Somalian biogeographical regions. We used 224 pancake tortoise occurrences obtained from Tanzania, Kenya and Zambia to estimate suitable and stable areas for the pancake tortoise in all countries present in these regions. We also used a protected area network to assess how many of the suitable and stable areas are protected for the conservation of this critically endangered species. Our model predicted the expansion of climatically suitable habitats for pancake tortoises from four countries and a total area of 90,668.75 km2 to ten countries in the future and an area of 343,459.60 - 401,179.70 km2. The model also showed that a more significant area of climatically suitable habitat for the species lies outside of the wildlife protected areas. Based on our results, we can predict that pancake tortoises may not suffer from habitat constriction. However, the species will continue to be at risk from the international pet trade, as most of the identified suitable habitats remain outside of protected areas. We suggest that efforts to conserve the pancake tortoise should not only focus on protected areas but also areas that are unprotected, as these comprise a large proportion of the suitable and stable habitats available following predicted future climate change.

ecology↗