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Home / Mitochondrial Multiplex Real-time Pcr as a Source Tracking Method in Fecal-contaminated Effluents

Mitochondrial Multiplex Real-time Pcr as a Source Tracking Method in Fecal-contaminated Effluents

Field assessment of horse-associated genetic markers HoF597 and mtCytb for detecting the source of contamination in surface waters

Abstract

We investigated the specificity and sensitivity of two horse-associated markers, HoF597 and Horse mtCytb, and 12 mitochondrial and bacterial markers of six animal species (human, cow, pig, bird, dog, chicken) in the faecal samples of 50 individual horses. Both horse markers were detected in 48 (96%) faecal samples. Cross-reactivity with dog (BacCan545) and pig (P23-2) occurred in 88% and 72% of horse faecal samples, respectively. Several other bacterial and mitochondrial markers of non-target hosts were also detected; however, their specificities were >80%. Analyses of samples from surface waters (n = 11) on or adjacent to properties from which horse faecal samples had been collected showed only the presence of HoF597 but not horse mitochondrial marker. Our data suggest that while bacterial and (or) mitochondrial markers of other animal species may be present in horse faeces, dog and pig markers may predominantly be present in horse faecal samples, which points to their nonspecificity as markers for microbial source tracking. Although HoF597 and Horse mtCytb are highly sensitive and specific for the detection of horse faecal pollution, because of their low numbers, mitochondrial (mtDNA) markers may not be robust for screening surface waters.

Résumé

Les auteurs ont étudié la spécificité et la sensibilité de deux marqueurs associés au cheval, HoF597 et le mtCytb équin, et 12 marqueurs mitochondriaux et bactériens de six espèces animales (humain, vache, porc, oiseau, chien, poulet) dans des échantillons fécaux de 50 chevaux. Les deux marqueurs équins ont été détectés dans 48 (96 %) des échantillons fécaux. Une réactivité croisée avec le chien (BacCan545) et le porc (P23-2) était détectée dans 88 % et 72 % des échantillons fécaux de cheval, respectivement. Plusieurs autres marqueurs bactériens et mitochondriaux d'hôtes non ciblés ont aussi été détectés, mais leur spécificité était supérieure à 80 %. Les analyses des échantillons des eaux de surface (n = 11) sur ou à proximité des propriétés sur lesquelles les échantillons fécaux de cheval ont été prélevés montraient la présence de HoF597 uniquement, mais pas du marqueur mitochondrial équin. Ces données suggèrent que si des marqueurs bactériens ou mitochondriaux d'autres espèces animales peuvent être présents dans les fèces du chevant, les marqueurs du chien et du porc peuvent être principalement présents dans les échantillons fécaux du cheval, ce qui indique leur non-spécificité en tant que marqueurs pour le dépistage des sources microbiennes. Même si HoF597 et le mtCytb équin sont hautement sensibles et spécifiques pour la détection de la pollution fécale des chevaux, en raison de leur faible nombre, les marqueurs mitochondriaux (ADNmt) pourraient ne pas être robustes pour le dépistage des eaux de surface. [Traduit par la Rédaction]

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Supplementary Material

Supplementary data (cjm-2019-0499suppla.zip)

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Published In

Canadian Journal of Microbiology cover image

Canadian Journal of Microbiology

Volume 66 • Number 11 • November 2020

History

Received: 25 September 2019

Revision received: 19 February 2020

Accepted: 21 February 2020

Published online: 21 July 2020

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© 2020.

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Key Words

  1. field assessment
  2. HoF597 marker
  3. microbial source tracking
  4. mtCytb
  5. surface waters

Mots-clés

  1. évaluation sur le terrain
  2. marqueur HoF597
  3. dépistage des sources microbiennes
  4. mtCytb
  5. eaux de surface

Authors

Affiliations

Jessica Gray

Genecology Research Centre, School of Health and Sport Sciences, University of the Sunshine Coast, Maroochydore DC 4558, Queensland, Australia.

Nicole Masters

Genecology Research Centre, School of Health and Sport Sciences, University of the Sunshine Coast, Maroochydore DC 4558, Queensland, Australia.

Aaron Wiegand

School of Science and Engineering, University of the Sunshine Coast, Maroochydore DC 4558, Queensland, Australia.

Genecology Research Centre, School of Health and Sport Sciences, University of the Sunshine Coast, Maroochydore DC 4558, Queensland, Australia.

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Mitochondrial Multiplex Real-time Pcr as a Source Tracking Method in Fecal-contaminated Effluents

Source: https://cdnsciencepub.com/doi/abs/10.1139/cjm-2019-0499