EquiNectar research Microbiome & metabolomics Peer reviewed EQN-S02

EquiNectar Spring Pasture Metabolomics Study

In a randomized study of 45 pastured horses and ponies, six weeks of EquiNectar supplementation produced a fecal volatile-metabolome profile that differed significantly from horses exposed to spring pasture without EquiNectar.

Original study title

Modulation of the equine microbiome by pasture and feed supplements: A metabolomics approach

Metabolome Volatile organic compounds Spring pasture Spring pasture Grazing

Study at a glance

Year 2019

Study type Randomized controlled field study

Sample 45 randomized: 24 EquiNectar, 21 control

Duration 6 weeks

Population Clinically healthy pastured horses and ponies aged 4–23 years, representing multiple breeds and types

Publication Peer-reviewed journal article

EquiNectar tested? Yes

Peer reviewed Yes

Open access Yes

Relationship EquiNectar research

Horses and ponies randomized

24 / 21

EquiNectar / control allocation

6 weeks

Spring pasture intervention period

p < 0.05

Difference between EquiNectar and control fecal VOC profiles at week six

Research question

What did the researchers want to understand?

How does six weeks of spring pasture alter the fecal volatile metabolome of horses and ponies, and does supplementation with EquiNectar produce an additional, distinguishable effect on that metabolomic response?

Researchers

Study authors

K. L. Snalune

MRCVS

Newnham Court Equine Clinic

John O. Hunter

Professor

Tharos Ltd
Addenbrooke's Hospital

Rosemary Waring

University of Birmingham
Tharos Ltd

Michael Cauchi

PhD

University of Limerick

C. Turner

Open University

C. Batty

Open University

Research organizations

Research institutions

Newnham Court Equine Clinic

Equine veterinary practice
United Kingdom

Addenbrooke's Hospital

Hospital
United Kingdom

University of Birmingham

University

University of Limerick

University
Ireland

Open University

University
United Kingdom

Methodology

How the study was conducted

Study design

Forty-five clinically healthy horses and ponies living at Redwings Sanctuary in Norfolk were enrolled during spring. The population comprised 36 geldings and nine mares, aged 4–23 years, representing a range of breeds including Shetland ponies, Welsh ponies, cob types and Thoroughbred or Thoroughbred-cross horses.

Animals were randomly allocated to a control group of 21 or an EquiNectar group of 24.

All animals had unrestricted access to pasture and received 200 g of a high-fiber ration twice daily. The EquiNectar group additionally received 300 ml of EquiNectar twice daily for six weeks.

Fecal samples were collected at baseline and after six weeks. One control horse was removed during the study after requiring treatment for a corneal ulcer.

Sampling and metadata collection were conducted under University of Cambridge ethics approval CR83 with the informed consent of the sanctuary.

What was measured

Fresh fecal samples were collected before the intervention and again after six weeks.

The volatile compounds present in the fecal headspace were analyzed using selected ion flow tube mass spectrometry (SIFT-MS). Samples were stored at −80°C before analysis.

The resulting mass-spectrometry data were analyzed using multivariate statistical techniques, including principal component analysis and classification models, to determine whether metabolomic profiles differed between treatment groups and between baseline and week six.

Body condition score and cresty neck score were also recorded before and after the intervention.

Study findings

Key results

• 45 horses and ponies were randomly allocated to EquiNectar (24) or control (21) groups.

• Both groups experienced substantial changes in their fecal volatile metabolome during six weeks on spring pasture.

• The EquiNectar group's fecal metabolome changed substantially between baseline and week six, with 78.7% classification accuracy.

• At week six, the EquiNectar and control groups had significantly different fecal VOC profiles, with 80.4% classification accuracy and an AUROC of 0.8569 (p<0.05).

• Acetate fell significantly in the EquiNectar group but not in the control group.

• Changes in methanol and acetone were observed following EquiNectar supplementation, although these individual differences were not statistically significant.

• Body condition score remained unchanged in both groups over the six-week study.

Interpretation

What do the findings mean?

Spring pasture had a substantial effect on the fecal metabolome, but EquiNectar modified that response.

At baseline, the EquiNectar and control groups could not be strongly distinguished by their fecal VOC profiles. After six weeks, the two groups were significantly different, with a classification accuracy of 80.4% and an AUROC of 0.8569.

Within the EquiNectar group, the week-zero and week-six metabolomes were also clearly distinguishable, with a classification accuracy of 78.7% and AUROC of 0.8875.

Importantly, the control group also changed markedly during the six weeks, demonstrating the strong influence of spring pasture. The study therefore does not suggest that every observed metabolic change was caused by EquiNectar. Instead, it shows that EquiNectar produced a distinguishable additional effect against a changing dietary background.

In practice

Why this research matters

Spring pasture can change substantially over a relatively short period, including changes in its non-structural carbohydrate content. This makes it difficult to separate the effects of a digestive supplement from the effects of changing grass unless an untreated comparison group is included.

This study addressed that problem by randomly allocating horses and ponies sharing the same spring grazing environment to either an EquiNectar group or a control group.

Both groups experienced substantial changes in their fecal volatile metabolome over six weeks, confirming that spring pasture itself had an important effect. However, by week six the EquiNectar and control groups also had significantly different metabolomic profiles.

The study therefore suggests that EquiNectar can influence the products of hindgut fermentation even against the background of substantial seasonal dietary change.

Research transparency

Study limitations

The study evaluated fecal volatile organic compounds rather than directly sequencing the microbial community. VOC profiles provide information about microbial metabolic activity, but they do not directly identify the abundance or composition of individual bacterial populations.

The trial lasted six weeks and was conducted at a single equine sanctuary during spring, so the results may not fully represent horses under different management systems, seasons or diets.

Although 45 horses and ponies were randomized, one control horse was subsequently removed after requiring treatment for a corneal ulcer.

The study population included a wide range of breeds, ages and body types. This improves real-world relevance but also introduces greater biological variation than would be present in a highly standardized experimental population.

Changes in the fecal metabolome should not be interpreted as direct measurements of conditions within the cecum or colon, or as evidence of treatment of a specific gastrointestinal disease.

EquiNectar context

How this research relates to EquiNectar

This study directly evaluated EquiNectar in a randomized population of horses and ponies living on spring pasture.

Unlike the earlier Thoroughbred study, this trial included a contemporaneous control group. Twenty-four animals received EquiNectar while 21 were allocated to the control group, allowing the researchers to distinguish changes associated with spring grazing from those associated with EquiNectar supplementation.

After six weeks, the fecal volatile-metabolome profile of the EquiNectar group differed significantly from that of the control group, providing direct evidence that EquiNectar influenced the downstream fermentation environment beyond the changes associated with spring pasture alone.

Tharos / EquiNectar involvement

This study directly evaluated EquiNectar. In the published paper the supplement is described as a commercially available Enzyme Rich Malt Extract (ERME), and the authors state that the same supplement had been used in the earlier Proudman et al. Thoroughbred study. The publication does not provide a separate funding statement or describe any additional role for Tharos Ltd.

EquiNectar evaluated Yes

Source record

Publication details

Journal

Integrative Food, Nutrition and Metabolism

Publisher

Open Access Text (OAT)

Year

2019

Volume

Pages / article

1–4

DOI

10.15761/IFNM.1000247

Species

Horse

Population

Clinically healthy pastured horses and ponies aged 4–23 years, representing multiple breeds and types

Sample

45 randomized: 24 EquiNectar, 21 control

Duration

6 weeks

Setting

Redwings Horse Sanctuary, Norfolk, UK, during spring pasture

License

CC BY 4.0

View original source ↗ Read full text ↗

Recommended citation

Snalune KL, Hunter JO, Waring RH, Cauchi M, Turner C, Batty C. Modulation of the equine microbiome by pasture and feed supplements: A metabolomics approach. Integrative Food, Nutrition and Metabolism. 2019;6:1–4. doi:10.15761/IFNM.1000247.

EquiNectar Spring Pasture Metabolomics Study

What did the researchers want to understand?

Study authors

K. L. Snalune

John O. Hunter

Rosemary Waring

Michael Cauchi

C. Turner

C. Batty

Research institutions

Newnham Court Equine Clinic

Addenbrooke's Hospital

University of Birmingham

University of Limerick

Open University

How the study was conducted

Key results

What do the findings mean?

Why this research matters

Study limitations

How this research relates to EquiNectar

Publication details

Related research

Racehorse Microbiome & Metabolome Study

EquiNectar Racehorse Fecal pH Study

Related practical resources

Hindgut Acidosis in Horses: Signs, Causes & Management