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Ecological and methodological insights from genetic and coprological profiling of gastrointestinal communities in wild howler monkeys


Abstract

The gastrointestinal tract hosts a complex community of microorganisms and helminth parasites that collectively contribute to host health and fitness. Analysis of these communities provides insight into diverse aspects of host dietary ecology, immunity, nutrition, and host-parasite interactions. However, research methodologies, such as sample preservation and sequencing approach, can influence how we understand and characterize these features. Here, we profiled the gastrointestinal microbial and helminth communities in different groups of wild Costa Rican mantled howler monkeys (Alouatta palliata palliata). We compared samples stored in ethanol versus directly flash frozen, and contrasted conclusions drawn from 16S versus shotgun sequencing approaches. Bacterial, archaeal, and eukaryotic taxa associated with the digestion of plant material dominated the GI communities. Storage and sequencing methods influenced microbial profiles: ethanol-stored samples exhibited higher diversity than frozen samples, and 16S sequencing detected lower diversity than shotgun. Helminths were detected via coprological microscopy in 71% of individuals, whereas metagenomic detection was inconsistent. This study provides new data on the microorganisms and their putative digestive functions in the gut of a folivorous primate, and highlights the pros and cons of different methodological choices when profiling host-microbiome and host-parasite interactions.

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Acknowledgements

We thank the administration team in Sector Santa Rosa for the assistance in the logistics and the support of our project, especially Roger Blanco and Maria Martha Chavarria. We would like to thank Gwen Duytschaever, Shasta E. Webb, and Matthew Workentine for their support with wet laboratory work. We further thank Megan C. Henriquez and Hadjira Hamou for providing feedback on an early version of the manuscript and on parasite identification.

Funding

This project was supported by the Natural Sciences and Engineering Research Council of Canada (RGPIN-2017-03782), the Canada Research Chairs Program (950-231257) and the University of Calgary (S.C., C.M., A.G.B, S.K., A.D.M). 

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Correspondence to
Silvia Carboni.

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The authors declare no competing interests.

Ethical

This study followed the guidelines of the Canadian Council of Animal Care and was approved by the Animal Care Committee (AC15-0161) of the University of Calgary, by the Sistema Nacional de Área de Conservación (SINAC R-ACG-PI-004-2017), and by the Comisión Nacional para la Gestión de la Biodiversidad (R-025-2014-OT-CONAGEBIO and R-029-2017-OT-CONAGEBIO) in Costa Rica. Samples were imported into Canada under permits issued by the Canadian Food Inspection Agency (CFIA , A-2016-03992-4).

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Carboni, S., Macfarland, C., Cheves Hernandez, S. et al. Ecological and methodological insights from genetic and coprological profiling of gastrointestinal communities in wild howler monkeys.
Sci Rep (2026). https://doi.org/10.1038/s41598-026-57628-6

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  • DOI: https://doi.org/10.1038/s41598-026-57628-6

Keywords

  • Parasite
  • Microbial ecology
  • Microbiome
  • Howler monkeys
  • Shotgun metagenomics
  • 16S amplicon sequencing


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