Testing storage methods of faecal samples for subsequent measurement of helminth egg numbers in the domestic horse.
Abstract: Parasite infection status, intensity and resistance have traditionally been quantified via flotation techniques, but the need for immediate analysis following defecation imposes limitations and has led to the use of several faecal storage techniques. However, their effect on nematode egg counts has not been systematically validated and is often generalised across taxa despite evidence of differences between species. Here, we take the domestic horse Equus ferus caballus as a model to examine the impact of commonly used storage techniques on egg recovery: 1) high and low concentrations of ethanol and formalin fixative solutions for up to four weeks and 2) refrigeration (3-5°C) over a two-week period. We found a significant decline in faecal egg counts (FEC) following storage in high and low concentrations of both fixative solutions after two weeks, which stabilised after four weeks, and this pattern was uniform across replicates. FECs remained relatively stable over a week of refrigeration, but declined when refrigeration exceeded 8 days. Prior to FEC analysis, we recommend sample refrigeration for no more than one week. Storage in either fixative solution is sub-optimal for the preservation of nematode eggs, although the uniformity of the decline across samples could hold potential for projective calculation of parasite egg shedding when storage time is effectively controlled for.
Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.
Publication Date: 2016-03-16 PubMed ID: 27084484DOI: 10.1016/j.vetpar.2016.03.012Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates the impact of different storage techniques on the measurement of helminth egg count in horse faeces. The paper concludes that refrigeration for no more than one week is recommended before analysis, and that traditional fixative solutions are sub-optimal for egg preservation, though they could be used with adjustments for known decline rates.
Understanding the Importance of the Study
- This study aims to find out how different storage techniques affect the ability to measure helminth egg counts in horse faeces. Helminths are parasitic worms, and researchers need to observe the number of eggs in the animal’s faeces to understand the parasite’s infection rate, intensity, and the host’s resistance.
- The traditional way to study parasite egg numbers is via flotation techniques, but these require immediate analysis post-defecation. This immediacy isn’t always achievable, hence the need for storage options. However, the effects of these storage techniques on egg count have not been systematically validated, which leaves room for inaccurate or inconsistent data.
Research Methodology
- Using the domestic horse as its model, the research studied two main storage options. The first one is the use of high and low concentrations of ethanol and formalin solutions, and the second one is refrigeration at 3-5°C. They explored these methods’ effects over periods up to four weeks and over a two-week period, respectively.
Research Findings
- The study found that storing faecal samples in both fixative solutions for two weeks significantly reduced the faecal egg count (FEC). This reduction stabilized after four weeks, and the pattern was consistent across all tested samples.
- While egg counts remained stable when refrigerated for a week, a decline was noted when refrigeration exceeded 8 days. Hence, the study suggests that fecal samples should be refrigerated for no more than a week before analysis.
Recommendations
- Based on the observations, the researchers suggested that neither high nor low concentrations of ethanol or formalin solutions are optimal for preserving nematode eggs for egg count analysis due to the noted decline in egg count. Yet, these storage methods could still be used, considering the consistent pattern of egg count decrease they observed across all samples over time. Researchers may adjust their measurements to account for this known rate of decline when analyzing sample data.
- The researchers found refrigeration to be a superior storage method, recommending it for no longer than a week before conducting FEC analysis. Beyond a week, the decline in egg count can give altered data.
Cite This Article
APA
Crawley JA, Chapman SN, Lummaa V, Lynsdale CL.
(2016).
Testing storage methods of faecal samples for subsequent measurement of helminth egg numbers in the domestic horse.
Vet Parasitol, 221, 130-133.
https://doi.org/10.1016/j.vetpar.2016.03.012 Publication
Researcher Affiliations
- Department of Animal and Plant Sciences, Alfred Denny Building, University of Sheffield, Western Bank, Sheffield S10 2TN, UK. Electronic address: jahcrawley1@gmail.com.
- Department of Animal and Plant Sciences, Alfred Denny Building, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
- Department of Animal and Plant Sciences, Alfred Denny Building, University of Sheffield, Western Bank, Sheffield S10 2TN, UK; Department of Biology, University of Turku, FIN-20014 Turku, Finland.
- Department of Animal and Plant Sciences, Alfred Denny Building, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
MeSH Terms
- Animals
- Ethanol / pharmacology
- Feces / parasitology
- Fixatives / pharmacology
- Formaldehyde / pharmacology
- Helminthiasis, Animal / diagnosis
- Helminthiasis, Animal / parasitology
- Helminths / drug effects
- Helminths / physiology
- Horses
- Parasite Egg Count / standards
- Parasite Egg Count / veterinary
- Refrigeration / standards
- Specimen Handling / methods
- Specimen Handling / standards
Citations
This article has been cited 18 times.- Nishigaki A, Previdelli R, Alexander JL, Balarajah S, Roberts L, Marchesi JR. Identification of a Sub-Clinical Salmonella spp. Infection in a Dairy Cow Using a Commercially Available Stool Storage Kit. Animals (Basel) 2023 Sep 4;13(17).
- Fontoura-Gonçalves C, Portocarrero É, Oliveira A, Lozano J, Rinaldi L, Cringoli G, Madeira de Carvalho L, Santos N. Challenges of copro-parasitological surveys in wild Iberian ibex (Capra pyrenaica) populations addressed through a combination of molecular and statistical tools. Ecol Evol 2023 Jun;13(6):e10172.
- Sy I, Conrad L, Becker SL. Recent Advances and Potential Future Applications of MALDI-TOF Mass Spectrometry for Identification of Helminths. Diagnostics (Basel) 2022 Dec 3;12(12).
- Abdu S, Chimento M, Alarcón-Nieto G, Zúñiga D, Aplin LM, Farine DR, Brandl HB. The performance of field sampling for parasite detection in a wild passerine. Ecol Evol 2022 Aug;12(8):e9242.
- Morgan ER, Lanusse C, Rinaldi L, Charlier J, Vercruysse J. Confounding factors affecting faecal egg count reduction as a measure of anthelmintic efficacy. Parasite 2022;29:20.
- De Noia M, Poole R, Kaufmann J, Waters C, Adams C, McGinnity P, Llewellyn M. Towards an in situ non-lethal rapid test to accurately detect the presence of the nematode parasite, Anguillicoloides crassus, in European eel, Anguilla anguilla. Parasitology 2022 Apr;149(5):605-611.
- Lobos-Ovalle D, Navarrete C, Navedo JG, Peña-Espinoza M, Verdugo C. Improving the sensitivity of gastrointestinal helminth detection using the Mini-FLOTAC technique in wild birds. Parasitol Res 2021 Sep;120(9):3319-3324.
- Gecchele LV, Pedersen AB, Bell M. Fine-scale variation within urban landscapes affects marking patterns and gastrointestinal parasite diversity in red foxes. Ecol Evol 2020 Dec;10(24):13796-13809.
- Ahmed N, Heitlinger E, Affinass N, Kühl AA, Xenophontos N, Jarquin VH, Jost J, Steinfelder S, Hartmann S. A Novel Non-invasive Method to Detect RELM Beta Transcript in Gut Barrier Related Changes During a Gastrointestinal Nematode Infection. Front Immunol 2019;10:445.
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- Mirzaei A, Rahmani Shahraki A, Maunsell FP, Diehl BN. Evaluation of the Diagnostic Performance and Validation of an AI-Assisted Fluorescence Imaging Device for Fecal Egg Counts Against the Manual McMaster Reference Method in Kiko Male Goats. Animals (Basel) 2026 Jan 14;16(2).
- van de Ven M, Broecke BV, Ribas A, Leirs H, Sabuni C, Mariën J. Efficiency of anthelmintic treatment and its effect on microparasite dynamics in wild Mastomys natalensis. Int J Parasitol Parasites Wildl 2025 Aug;27:101098.
- Prescott K, Michels E, Tschirren B. Co-Infection, but Not Infection Intensity, Increases Shedding in a Gastrointestinal Helminth of Gamebirds. Ecol Evol 2025 Jun;15(6):e71483.
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- Lee YJ, Phang GJ, Chen CC, Ou JH, Fan YH, Huang YT. Optimal liquid-based DNA preservation for DNA barcoding of field-collected fungal specimens. Heliyon 2024 Sep 15;10(17):e36829.
- Rijal S, Neuhaus P, Thorley J, Caulkett N, Kutz S, Ruckstuhl KE. Patterns of gastrointestinal parasite infections in bighorn sheep, Ovis canadensis, with respect to host sex and seasonality. Int J Parasitol Parasites Wildl 2024 Aug;24:100950.
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