The equine gastrointestinal microbiome: impacts of weight-loss.

BACKGROUND: Obesity is an important equine welfare issue. Whilst dietary restriction is the most effective weight-loss tool, individual animals range in their weight-loss propensity. Gastrointestinal-derived bacteria play a fundamental role in host-health and have been associated with obesity and weight-loss in other species. This study evaluated the faecal microbiome (next-generation sequencing of 16S rRNA genes) of 15 obese Welsh Mountain pony mares, in the same 11-week period across 2 years (n = 8 Year 1; n = 7 Year 2). Following a 4-week acclimation period (pre-diet phase) during which time individuals were fed the same hay to maintenance (2% body mass (BM) as daily dry matter (DM) intake), animals underwent a 7-week period of dietary restriction (1% BM hay as daily DM intake). Faeces were sampled on the final 3 days of the pre-diet phase and the final 3 days of the dietary restriction phase. Bacterial communities were determined using Next Generation Sequencing of amplified V1-V2 hypervariable regions of bacterial 16S rRNA. RESULTS: Losses in body mass ranged from 7.11 to 11.59%. Changes in the faecal microbiome composition following weight-loss included a reduction in the relative abundance of Firmicutes and Tenericutes and a reduction in indices of bacterial diversity. Pre-diet diversity was negatively associated with weight-loss. Pre-diet faecal acetate concentration was a strong predictor of subsequent weight-loss and negatively associated with Sphaerochaeta (Spirochaetes phylum) abundance. When animals were divided into 3 groups (high, mid, low) based overall weight loss, pre-diet bacterial community structure was found to have the greatest divergence between the high and low weight-loss groups (R = 0.67, p <  0.01), following PERMANOVA and ANOSIM analysis. CONCLUSIONS: Weight-loss in this group of ponies was associated with lower pre-diet faecal bacterial diversity and greater pre-diet acetate concentration. Overall, these data support a role for the faecal microbiome in weight-loss propensity in ponies and provide a baseline for research evaluating elements of the faecal microbiome in predicting weight-loss success in larger cohorts.

In vitro susceptibility of contagious ovine digital dermatitis associated Treponema spp. isolates to antimicrobial agents in the UK.

BACKGROUND: Contagious ovine digital dermatitis (CODD) is an important cause of infectious lameness in sheep in the UK and Ireland and has a severe impact on the welfare of affected individuals. The three treponemal phylogroups Treponema medium/Treponema vincentii-like, Treponema phagedenis-like and Treponema pedis spirochaetes have been associated with clinical CODD lesions and are considered to be a necessary cause of disease. There are scant data on the antimicrobial susceptibility of the treponemes cultured from CODD lesions. OBJECTIVE: The aim of this study was to determine in vitro the miniumum inhibitory concentration/ minimum bactericidal concentration (MIC/MBC) of antimicrobials used in the sheep industry for isolates of the three CODD associated treponeme phylogroups T. medium/T. vincentii-like, T. phagedenis-like and T. pedis. ANIMALS: Twenty treponeme isolates; from 19 sheep with clinical CODD lesions. METHODS: A microdilution method was used to determine in vitro the MIC/MBC of 10 antimicrobial agents for 20 treponeme isolates (five T. medium/T. vincentii-like, 10 T. phagedenis-like and five T. pedis). The antimicrobials tested were penicillin G, amoxicillin, oxytetracycline, tilmicosin, lincomycin, spectinomycin, tylosin, tildipirosin, tulathromycin and gamithromycin. RESULTS: The treponeme isolates tested showed low MICs and MBCs to all 10 antimicrobials tested. They were most susceptible to gamithromycin and tildipirosin (MIC90: 0.0469 mg/L), and were least susceptible to lincomycin, spectinomycin and oxytetracycline (MIC90: 48 mg/L, 24 mg/L and 3 mg/L, respectively). CONCLUSIONS: These data are comparable to in vitro antimicrobial susceptibility data for treponemes cultured from bovine digital dermatitis lesions. Dependent on local licensing, penicillin and tilmicosin appear to be the best candidates for future in vivo studies.

Comparison of clinical cure rates from footrot and contagious ovine digital dermatitis using zinc sulphate foot bathing and topical oxytetracycline: A randomised trial.

BACKGROUND: This study reports the clinical cure rates of topical oxytetracycline and 10% zinc sulphate foot bathing for treatment of interdigital dermatitis (ID), footrot (FR) and contagious ovine digital dermatitis (CODD) in lambs. METHODS: The study was a randomised controlled trial of 75 lambs. Group A (n = 38) was foot bathed daily for 5 days in 10% zinc sulphate for 15 minutes and group B was treated with daily topical oxytetracycline for 5 days. On days 0, 7, 14, 28 and 42, lambs were scored for locomotion and foot lesions were recorded. RESULTS: The initial cure rates for ID were 96.20% and 97.00%; for FR, 100% and 95%; and for CODD, 90.09% and 83.33% for zinc sulphate and oxytetracycline, respectively. By day 42, these had changed to 53.16% and 61% for ID; 47.82% and 70% for FR; and 100% and 83.33% for CODD. There were no significant differences in cure rates between the treatments for most time points. LIMITATIONS: The sample size was small, and further studies in larger cohorts and different classes of sheep are required before the findings can be translated into recommendations for clinical practice. CONCLUSION: Both treatments achieved cure rates that are comparable to reported cure rates using systemic antibiotics and could be an effective alternative.

COVID-19 impacts equine welfare: Policy implications for laminitis and obesity.

The COVID-19 pandemic continues to impact human health and welfare on a global level. In March 2020, stringent national restrictions were enforced in the UK to protect public health and slow the spread of the SARS-CoV-2 virus. Restrictions were likely to have resulted in collateral consequences for the health and welfare of horses and ponies, especially those at risk of obesity and laminitis and this issue warranted more detailed exploration. The current study utilised qualitative methodology to investigate the implications of COVID-19 related policies upon equine management and welfare with a focus on horses and ponies at risk of laminitis and obesity. Twenty-four interviews with five sub-groups of key equestrian welfare stakeholders in the UK were conducted between May and July of 2020 to understand the challenges facing equine welfare in the context of laminitis and obesity susceptible animals. Thematic analysis revealed lockdown-associated factors with the potential to compromise welfare of horses and ponies at risk of obesity and laminitis. These included: disparate information and guidance, difficulties enacting public health measures in yard environments, and horses having reduced exercise during the pandemic. Positive examples of clear and consistent information sharing by farriers were reported to have improved horse owner understanding of routine hoof care during lockdown. Analysis suggested that the recommendations for supporting the management-based needs of horses under reduced supervision were not clearly defined, or were not sufficiently disseminated, across the equine industry. These findings support the development of guidelines specific to the care of horses and ponies at risk of obesity and laminitis through collaborative input from veterinary and welfare experts, to reduce the negative impacts of future lockdown events in the UK.

Repeatability and Feasibility of Pressure Algometry for Quantifying Mechanical Nociceptive Threshold in the Thoracic Region of Calves.

Pressure algometry can be used to quantify mechanical nociceptive threshold (MNT) in humans and animals. If reliable this may be a useful tool to examine calves for increased mechanical sensitivity, which may be induced by disease or pain. This study measures the repeatability and feasibility of pressure algometry using a handheld digital pressure algometer (PRODPlus, Top Cat metrology) using three serial measurements applied to six sites on the thoraces of 35 healthy calves by two different operators. The range of MNTs recorded in healthy calves was 1.2-25 Newtons (median = 10.1 IQR = 7.1-14.0). A multivariable mixed effects model identified that the MNT's recorded were influenced by Operator, Site, and Calf. Intra and inter-operator reliability were measured by intra-class correlation coefficients (ICCs). Based on average ICCs, intra-operator reliability at two sites was good; one site overlying the ventral aspect of the 6th intercostal space [ICC = 0.79 95% CI (0.63-0.89)] and the other overlying the dorsal aspect of the 9th intercostal space [ICC = 0.75 95% CI (0.56-0.87)]. Average ICCs for three other measurement sites were moderate or poor, and one site proved unfeasible. For inter-operator agreement average ICCs showed that agreement was also good at the same 6 and 9th intercostal space, [ICCs = 0.77 95% CI (0.35-0.90) and 0.77 95% CI (0.54-0.88), respectively], agreement was moderate for the remainder of the sites. This study identifies two sites that are potentially useful for monitoring of thoracic sensitivity as an indicator of pain in calves by means of pressure algometry using the average of three measurements. It also identifies sources of variability to be considered when applying the tool for clinical or research purposes.

Effect of age and the individual on the gastrointestinal bacteriome of ponies fed a high-starch diet.

Bacteria residing in the gastrointestinal tract of mammals are crucial for the digestion of dietary nutrients. Bacterial community composition is modified by age and diet in other species. Although horses are adapted to consuming fibre-based diets, high-energy, often high-starch containing feeds are increasingly used. The current study assessed the impact of age on the faecal bacteriome of ponies transitioning from a hay-based diet to a high-starch diet. Over two years, 23 Welsh Section A pony mares were evaluated (Controls, 5-15 years, n = 6/year, 12 in total; Aged, ≥19 years, n = 6 Year 1; n = 5 Year 2, 11 in total). Across the same 30-week (May to November) period in each year, animals were randomly assigned to a 5-week period of study and were individually fed the same hay to maintenance (2% body mass as daily dry matter intake) for 4-weeks. During the final week, 2g starch per kg body mass (micronized steam-flaked barley) was incorporated into the diet (3-day transition and 5 days at maximum). Faecal samples were collected for 11 days (final 3 days hay and 8 days hay + barley feeding). Bacterial communities were determined using Ion Torrent Sequencing of amplified V1-V2 hypervariable regions of 16S rRNA. Age had a minimal effect on the bacteriome response to diet. The dietary transition increased Candidatus Saccharibacteria and Firmicutes phyla abundance and reduced Fibrobactres abundance. At the genera level, Streptococcus abundance was increased but not consistently across individual animals. Bacterial diversity was reduced during dietary transition in Streptococcus 'responders'. Faecal pH and VFA concentrations were modified by diet but considerable inter-individual variation was present. The current study describes compositional changes in the faecal bacteriome associated with the transition from a fibre-based to a high-starch diet in ponies and emphasises the individual nature of dietary responses, which may reflect functional differences in the bacterial populations present in the hindgut.

Sheep and farm level factors associated with footrot: a longitudinal repeated cross-sectional study of sheep on six farms in the UK.

Footrot is an ovine foot disease of infectious origin and a cause of serious welfare and economic compromise in affected animals and flocks. The development of footrot in sheep is associated with the infectious agent Dichelobacter nodosus, which may invade as a primary pathogen, but the risk of disease is increased following damage to the interdigital skin of the foot. In this study, we used data from six farms in North Wales collected between June 2012 and October 2013 to model the dynamic changes of footrot prevalence over time and investigate the association of footrot with multiple farm, management, environmental and sheep factors. Footrot prevalence varied widely within and between farms and overall varied with season with an increase in prevalence shown in late summer and again in the spring. In addition, sheep were more likely to have footrot when the flock size was larger, when grazing poached pasture or when grazing a longer sward, and yearling sheep were less likely to have footrot when compared with lambs and adult sheep. These data may be helpful for advising farmers of likely environmental events, risk groups and management practices that may increase the probability of sheep developing footrot.

The Equine Gastrointestinal Microbiome: Impacts of Age and Obesity.

Gastrointestinal microbial communities are increasingly being implicated in host susceptibilities to nutritional/metabolic diseases; such conditions are more prevalent in obese and/or older horses. This controlled study evaluated associations between host-phenotype and the fecal microbiome / metabolome. Thirty-five, Welsh Mountain pony mares were studied across 2 years (Controls, n = 6/year, 5-15 years, Body Condition Score (BCS) 4.5-6/9; Obese, n = 6/year, 5-15 years, BCS > 7/9; Aged, n = 6 Year 1; n = 5 Year 2, ≥19 years old). Animals were individually fed the same hay to maintenance (2% body mass as daily dry matter intake) for 2 (aged / obese) or 4 (control), 4-week periods in a randomized study. Outset phenotype was determined (body fat%, markers of insulin sensitivity). Feces were sampled on the final 3 days of hay feeding-periods and communities determined using Next Generation Sequencing of amplified V1-V2 hypervariable regions of bacterial 16S rRNA. Copy numbers for fecal bacteria, protozoa and fungi were similar across groups, whilst bacterial diversity was increased in the obese group. Dominant bacterial phyla in all groups were Bacteroidetes > Firmicutes > Fibrobacter. Significant differences in the bacterial communities of feces were detected between host-phenotype groups. Relative to controls, abundances of Proteobacteria were increased for aged animals and Bacteroidetes, Firmicutes, and Actinobacteria were increased for obese animals. Over 500 bacterial operational taxonomic units (OTUs) differed significantly between host-phenotype groups. No consistent pattern of changes in discriminant OTUs between groups were maintained across groups and between years. The core bacterial populations contained 21 OTUs, 6.7% of recovered sequences. Distance-based Redundancy Analyses separated fecal bacterial communities with respect to markers of obesity and insulin dysregulation, as opposed to age. Host-phenotype had no impact on the apparent digestibility of dietary GE or DM, fecal volatile fatty acid concentrations or the fecal metabolome (FT-IR). The current study demonstrates that host-phenotype has major effects on equine fecal microbial population structure. Changes were predominantly associated with the obese state, confirming an obesity-associated impact in the absence of nutritional differences. Clear biomarkers of animal-phenotype were not identified within either the fecal microbiome or metabolome, suggesting functional redundancy within the gut microbiome and/or metabolome.

Effect of Calf Gender on Milk Yield and Fatty Acid Content in Holstein Dairy Cows.

The scale of sexed semen use to avoid the birth of unwanted bull calves in the UK dairy industry depends on several economic factors. It has been suggested in other studies that calf gender may affect milk yield in Holsteins- something that would affect the economics of sexed semen use. The present study used a large milk recording data set to evaluate the effect of calf gender (both calf born and calf in utero) on both milk yield and saturated fat content. Linear regression was used to model data for first lactation and second lactation separately. Results showed that giving birth to a heifer calf conferred a 1% milk yield advantage in first lactation heifers, whilst giving birth to a bull calf conferred a 0.5% advantage in second lactation. Heifer calves were also associated with a 0.66kg reduction in saturated fatty acid content of milk in first lactation, but there was no significant difference between the genders in second lactation. No relationship was found between calf gender and milk mono- or polyunsaturated fatty acid content. The observed effects of calf gender on both yield and saturated fatty acid content was considered minor when compared to nutritional and genetic influences.