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Record W4414709211 · doi:10.1101/2025.09.29.679195

Introducing NOMAD: A Field-Deployable Workflow for Rapid, On-Site Microbiome Analysis of Raw Bovine Milk Using Full-Length 16S rRNA Sequencing

2025· preprint· en· W4414709211 on OpenAlexaff
Valeria R. Parreira, Melinda Precious, Gurleen Taggar, Dharamdeo Singh, Lawrence Goodridge

Bibliographic record

VenuebioRxiv (Cold Spring Harbor Laboratory) · 2025
Typepreprint
Languageen
FieldAgricultural and Biological Sciences
TopicMilk Quality and Mastitis in Dairy Cows
Canadian institutionsUniversity of Guelph
Fundersnot available
KeywordsMicrobiomeMetagenomicsWorkflowMinionNanopore sequencingRaw milkDNA extraction

Abstract

fetched live from OpenAlex

ABSTRACT Raw bovine milk hosts a diverse microbiota that profoundly influences dairy product quality, safety, and shelf-life. However, current surveillance methods are time-intensive and often lack the taxonomic resolution needed for effective risk mitigation. To address this, we developed NOMAD (Nanopore-based On-site Microbiome Analysis of Diversity), a field-deployable workflow for rapid, high-resolution characterization of the raw milk microbiome using full-length 16S rRNA gene sequencing via Oxford Nanopore Technologies. Milk samples collected from a commercial dairy operation were processed using eight DNA extraction protocols, with Method (incorporation of EDTA and TE buffer) emerging as the optimal approach for microbial richness and DNA yield. Sequencing was performed on a MinION Mk1B platform, and bioinformatic analyses revealed that a 4-hour run was sufficient to recover >90% of total community richness, with stable alpha and beta diversity metrics by this timepoint. The complete workflow, including DNA extraction, library preparation, sequencing, and analysis, was completed in 10.5 hours, enabling same-day microbiome profiling in farm-adjacent settings. Comparative analysis showed strong agreement with established milk microbiome studies, while full-length reads enhanced resolution of spoilage-associated taxa such as Pseudomonas spp. and Streptococcus spp. The NOMAD platform offers a powerful and practical tool for near real-time microbiological surveillance in the dairy industry, supporting proactive quality control and improved food safety outcomes.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.002
metaresearch head score (Gemma)0.002
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Methods · Consensus signal: Methods
Teacher disagreement score0.003
Threshold uncertainty score0.013

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0020.002
Meta-epidemiology (narrow)0.0010.001
Meta-epidemiology (broad)0.0010.001
Bibliometrics0.0010.000
Science and technology studies0.0010.000
Scholarly communication0.0020.001
Open science0.0010.002
Research integrity0.0010.001
Insufficient payload (model declined to judge)0.0030.002

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.025
GPT teacher head0.237
Teacher spread0.212 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreMethods

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations0
Published2025
Admission routes1
Has abstractyes

Explore more

Same venuebioRxiv (Cold Spring Harbor Laboratory)→Same topicMilk Quality and Mastitis in Dairy Cows→French-language works237,207→