Molecular species determination of cyathostomins from horses in Ireland.

Cyathostomins are globally important equine parasites, responsible for both chronic and acute pathogenic effects. The occurrence of mixed infections with numerous cyathostomin species hinders our understanding of parasite epidemiology, host-parasite dynamics, and species pathogenicity. There have been few studies of cyathostomin species occurring in horses in Ireland, where temperate climatic conditions with year-round rainfall provide suitable conditions for infection of grazing animals with bursate nematodes. Here, we amplified and sequenced the ITS-2 region of adult worms harvested at post-mortem from eleven adult horses between August 2018 and June 2020, and recorded species prevalence and abundance of worms recovered from the caecum, right ventral colon and left dorsal colon, using both BLAST and IDTAXA for taxonomic attribution. Phylogenetic relationships and community composition were also recorded and compared with other relevant studies, including a global meta-analysis. Overall, our results agree with previous studies that there does not seem to be a major difference in cyathostomin species occurrence in equids in different geographical regions. We confirmed the results of other workers in relation to the difficulties in discriminating between Cylicostephanus calicatus and Coronocyclus coronatus on the basis of ITS-2 sequences.

First report of Besnoitia bennetti in Irish donkeys: an emerging parasitic disease in Europe.

BACKGROUND: This is the first report of Besnoitia bennetti in donkeys in Ireland. B. bennetti, an apicomplexan protozoan parasite specific to equids, is an emerging pathogen in Europe. This parasite forms chronic intracytoplasmic cysts in cells of the mesenchymal lineage, mainly fibroblasts, in the skin, sclera and mucosa. Clinical signs in affected equine hosts vary from mild to severe debilitating disease. Little is known of the phylogeny, epidemiology or transmission of B. bennetti infection in donkeys, mules or horses. CASE PRESENTATION: Two cases of besnoitiosis in donkeys are presented. Both donkeys were born and raised in theSouthwest of Ireland. The first case was a 2.5-year-old donkey that had a suspect sarcoid removed, while the second case,a 2-year-old donkey, had a biopsy of nodular dermatitis of the muzzle. Diagnosis was made by histopathology and the parasite species, B. bennetti, was confirmed by PCR followed by sequencing and microsatellite analysis. Both donkeys had high antibody titres against Besnoitia spp. Small (0.5 mm) scleral, conjunctival and dermal cysts over the muzzle were subsequently observed in both animals. Treatment with trimethoprim sulfadiazine for 30 days did not lead to clinical resolution. The findings are compared to the cases of besnoitiosis in donkeys reported in the past 10 years throughout Europe. CONCLUSIONS: Besnoitiosis should be considered as a differential diagnosis for chronic skin disease particularly in cases of cutaneous masses, non-pruritic dermatitis, and dermatitis that is not responsive to treatment in donkeys and other equids. Future studies are needed to investigate the prevalence of the disease in Irish donkeys, the spread of the disease and the potential impact on the health and welfare of the donkeys.

Validation of a serum ELISA test for cyathostomin infection in equines.

Cyathostomins are ubiquitous equine nematodes. Infection can result in larval cyathostominosis due to mass larval emergence. Although faecal egg count (FEC) tests provide estimates of egg shedding, these correlate poorly with burden and provide no information on mucosal/luminal larvae. Previous studies describe a serum IgG(T)-based ELISA (CT3) that exhibits utility for detection of mucosal/luminal cyathostomins. Here, this ELISA is optimised/validated for commercial application using sera from horses for which burden data were available. Optimisation included addition of total IgG-based calibrators to provide standard curves for quantification of antigen-specific IgG(T) used to generate a CT3-specific 'serum score' for each horse. Validation dataset results were then used to assess the optimised test's performance and select serum score cut-off values for diagnosis of burdens above 1000, 5000 and 10,000 cyathostomins. The test demonstrated excellent performance (Receiver Operating Characteristic Area Under the Curve values >0.9) in diagnosing infection, with >90% sensitivity and >70% specificity at the selected serum score cut-off values. CT3-specific serum IgG(T) profiles in equines in different settings were assessed to provide information for commercial test use. These studies demonstrated maternal transfer of CT3-specific IgG(T) in colostrum to newborns, levels of which declined before increasing as foals consumed contaminated pasture. Studies in geographically distinct populations demonstrated that the proportion of horses that reported as test positive at a 14.37 CT3 serum score (1000-cyathostomin threshold) was associated with parasite transmission risk. Based on the results, inclusion criteria for commercial use were developed. Logistic regression models were developed to predict probabilities that burdens of individuals are above defined thresholds based on the reported serum score. The models performed at a similar level to the serum score cut-off approach. In conclusion, the CT3 test provides an option for veterinarians to obtain evidence of low cyathostomin burdens that do not require anthelmintic treatment and to support diagnosis of infection.

Obesity and obesity-associated metabolic disease conditions in Connemara ponies in Ireland.

BACKGROUND: Equine obesity combined with insulin dysregulation (ID) is a major risk factor associated with laminitis. Some pony breeds appear to be at increased risk. However, little is known regarding the prevalence of obesity or hyperinsulinaemia as evidence of ID in Irish ponies. OBJECTIVE: To investigate the prevalence of obesity and associated endocrine/metabolic disease conditions in Connemara ponies and to determine if hyperinsulinaemia in these ponies could be predicted by morphometric or metabolic markers. STUDY DESIGN: Cross-sectional study. METHODS: The study population included registered Connemara ponies recruited through public and veterinary social media posts. Ponies underwent a physical examination and information on their management and clinical history was obtained via owner questionnaire. The body condition score (BCS) was measured using the Henneke system; cresty neck score (CNS) and regionalised adiposity were also assessed. Hyperinsulinaemia was confirmed by measuring serum basal insulin concentration (BIC) or insulin concentration after an oral sugar test (OST). Blood glucose and triglyceride concentrations were measured. Characteristics of hyperinsulinaemic and insulin-sensitive ponies were compared by logistic regression. RESULTS: Two hundred ponies were included; 59 ponies (29.5%) had a BCS ≥7, 58 (29.0%) had a CNS ≥2.5 and 135 (67.5%) had regionalised adiposity; 137 (68.5%) ponies had at least one of these abnormalities. Owner-reported history or clinical evidence of chronic laminitis was found in 92 ponies (46.0%). Hyperinsulinaemia was confirmed in 32 ponies (16.0%), including 23 of 91 (25.3%) detected by OST and 9 of 109 (8.3%) by BIC. Hypertriglyceridaemia was observed in 12 of 198 ponies (6.1%) ponies and hyperglycaemia in 11 of 197 ponies (5.6%) ponies. The odds of hyperinsulinaemia increased by a factor of 6.53 (95% confidence interval: 2.95, 15.21) when BCS was ≥7. MAIN LIMITATIONS: The OST was not performed in all ponies. CONCLUSIONS: Increased adiposity, laminitis and metabolic derangements are prevalent in this native Irish pony breed.

Rate of obesity within a mixed-breed group of horses in Ireland and their owners' perceptions of body condition and useability of an equine body condition scoring scale.

BACKGROUND: Equine obesity is a significant health and welfare concern. The proportion of domestic horse populations that are overweight are as high as 45%. As the primary decision-makers for their horses' care, owners are theoretically ideally placed to identify whether their horses are appropriately conditioned, however, research in other countries has shown that many owners are unable to accurately judge their horse's body condition. In this study, through the comparison of body condition scoring (BCS) performed by an expert and the horse owners and interviews with owners, we aimed to identify the proportion of horses that were overweight or obese, to assess the accuracy of horse owners' BCS assessment both prior to and after receiving information and instructions on body condition scoring, and to identify common themes amongst owners' views regarding BCS assessment and the Henneke BCS system. RESULTS: Forty-five percent of the horses in this study were overweight or obese. The agreement between the owners and an equine veterinarian regarding the horses' BCS was fair to good both prior to (κ = .311, P < 0.001; ICC = .502, P < 0.001) and after (κ = .381, P < 0.001; ICC = .561, P < 0.001) receiving information and instructions on scoring. Three quarters of the owners who took part in the study did not use any method of monitoring their horse's body condition. Thematic analysis of owner responses was varied, with the most common theme being an awareness of the need to monitor or make changes to their horse's condition with responses in this theme split between owners who felt in control and those who did not. Owner feedback on the utility and useability of the scorning system was that it was useful however parts are too technical or need improvement. CONCLUSIONS: Equine obesity is a significant problem in this population in Ireland. Horse owners' ability to accurately judge their horse's condition does not improve with provision of instructions on body condition scoring. These results combined with owners' feedback on the Henneke BCS system indicate that it is not a tool that can be reliably used by owners.

A qualitative study of perceived barriers and facilitators to sustainable parasite control on thoroughbred studs in Ireland.

In view of the rising threat of anthelmintic resistance in parasite populations and the planned change of classification of anthelmintics in Ireland for food animals to prescription-only medicines, there is a need for increased focus on parasite control strategies for horses. Effective parasite control programs (PCPs) are complex, necessitating the application of risk-assessment based on host immune status, infection pressure, type of parasite and seasonality to determine the requirement for anthelmintic administration and an understanding of parasite biology to employ effective, non- therapeutic, parasite control measures. The aim of this study was to explore horse breeders' attitudes and behaviours towards general parasite control practices and anthelmintic use on thoroughbred studs in Ireland using qualitative research methods, in order to help identify barriers to the implementation of sustainable equine PCPs supported by veterinary involvement. One-to-one, qualitative, semi-structured interviews were conducted with 16 breeders, using an interview topic guide that allowed for an open questioning style. The topic guide facilitated discussion around the following areas: (i) parasite control measures (general approach, (ii) veterinary involvement, (iii) use of anthelmintics, (iv) use of diagnostics, pasture management), (v) recording of anthelmintic use and (vi) anthelmintic resistance. Convenience, purposive (subjective selection) sampling was used to select a small group of participants that reflected the characteristics of current thoroughbred breeders in Ireland taking into account farm type, farm size, geographical location. The interviews were transcribed, and a method of inductive thematic analysis (a data driven identification and analysis of themes) was performed. Assessment of current behaviours by these participants found that PCPs were primarily based on the prophylactic use of anthelmintics without a strategic rationale. A key driver of behaviour regarding parasite prevention practices was localised routine, dictated by tradition, that gave breeders a sense of confidence and protection regarding parasite control. The perceptions of the benefits of parasitology diagnostics were varied and their application to control was poorly understood. Anthelmintic resistance was recognised as an industry threat but not an issue for individual farms. By using a qualitative approach, this study offers insights into the possible barriers to the adoption of sustainable PCPs on Irish thoroughbred farms and highlights the need for end-user involvement in the preparation of future guidelines.

Survey of the knowledge and perceptions of horse owners in Ireland of common clinical conditions and their impact.

BACKGROUND: As the primary decision-maker for their horse's health and welfare, owners' knowledge of clinical conditions may impact their horse's health. Anecdotal evidence suggests that the emotional impact of equine illness on the owner can be severe but research is lacking. OBJECTIVES: To evaluate horse owners' self-declared knowledge of eight common equine health conditions and perceptions of the quality of information available; to ascertain respondents' perceptions of the severity of impact on their horse of the conditions and potential emotional impact on themselves and to establish the factors of greatest concern to owners when their horse has a health condition and influential factors on end-of-life decisions. STUDY DESIGN: Online survey of horse owners. METHODS: An online survey comprising three sections was created: respondent demographics; vignettes of clinical conditions; concerns and decision-making regarding illness and euthanasia. Vignettes focused on arthritis; Pars Pituitary Intermedia Dysfunction; Equine Metabolic Syndrome (EMS); gastric ulcers; high intestinal worm burden; dermatophilosis ('mud rash'); quidding and head tossing; and equine asthma. Respondents were self-selecting. The survey was circulated for completion by horse owners residing in Ireland and Northern Ireland obtaining 491 valid responses. RESULTS: Respondents declared that they had the highest knowledge of mud rash, with 84.1% rating their knowledge as very good compared to 42% of respondents for EMS. There was positive correlation between the perceived impact on their horse and emotional impact on the owner for all conditions ranging from weak (arthritis 0.36, P < 0.001) to strong (EMS 0.62, P < 0.001). Pain, discomfort, and quality of life were reported as the most common concerns when their horse has a medical condition or when considering euthanasia. MAIN LIMITATIONS: Potential bias among respondents towards computer literate owners, with good literacy and interest in equine health conditions. CONCLUSIONS: Knowledge of equine health conditions, perceived impact of conditions on the horse and emotional impact on their owners varied significantly between conditions.

A Systematic Review of Arts-Based Interventions Delivered to Children and Young People in Nature or Outdoor Spaces: Impact on Nature Connectedness, Health and Wellbeing.

Background: The time that children and young people spend in nature and outdoor spaces has decreased significantly over the past 30 years. This was exacerbated with a further 60% decline post-COVID-19. Research demonstrating that natural environments have a positive impact on health and wellbeing has led to prescription of nature-based health interventions and green prescribing, although evidence for its use is predominantly limited to adults. Growing evidence also shows the impact of arts on all aspects of health and wellbeing. However, what has received scant attention in literature is the interconnection between the two: arts and nature. Aims: This review synthesizes the literature surrounding the interconnectedness between arts and nature, and their impact on the health and wellbeing of children and young people. Methods: Eight major electronic databases were systematically searched, while hand-searching included 20 journals, six books, and contact with experts. The review was conducted using the Cochrane handbook for systematic reviews, PRISMA guidelines and TIDieR template. All stages were conducted independently by two researchers and the protocol was published on PROSPERO (Registration no.: CRD42021286574). Results: Although 9,314 records were identified, only 11 records were included as most studies focused either on arts or nature, but not both. Studies were conducted in United Kingdom, United States, Ireland, Australia, and Hong Kong, in a range of spaces such as forests, woodlands, beaches, parks, fields, gardens, and school playgrounds. The review encompasses data from 602 participants in total. Discussion: Arts-in-nature offered an inclusive medium to engage all children and young people, especially those who might otherwise remain disinterested about environmental issues and disengaged with educational programs. Further, arts-in-nature provided stimuli to increase nature connectivity, understand environmental issues and explore ways to prevent environmental disasters. This led to higher environmental awareness and pro-environmental behaviors, and potential decrease in eco-anxiety. Conclusion: Although the quality of qualitative studies was high, the quality of quantitative studies was low or unclear, thus quantitative evidence is still at its infancy. Implications for research, policy, and practice are discussed, such as methods and activities to strengthen future interventions. Scaling-up existing interventions may lead to wider recognition and inclusion of arts-in-nature in future health guidelines, including green prescribing.

Assessment of Environmental and Occupational Risk Factors for the Mitigation and Containment of a COVID-19 Outbreak in a Meat Processing Plant.

Throughout the COVID-19 pandemic, meat processing plants have been vulnerable to outbreaks of SARS-CoV-2 infection. Transmission of the virus is difficult to control in these settings because of a combination of factors including environmental conditions and the specific nature of the work. This paper describes a retrospective outbreak investigation in a meat processing plant, a description of the measures taken to prevent or contain further outbreaks, and insights on how those with specific knowledge of the working environment of these plants can collaborate with public health authorities to ensure optimal outbreak control. The plant experienced 111 confirmed positive asymptomatic cases in total with an estimated attack rate of 38% during a five-week period. 4 weeks after the first case, mass screening of all workers was conducted by the public health authorities. Thirty-two workers tested positive, of which 16 (50%) worked in one particular area of the plant, the boning hall (n = 60). The research team prepared and carried out semi-structured interviews with the plant personnel who were charged with COVID control within the plant. They carried out assessments of operational risk factors and also undertook air quality monitoring in the boning hall and abattoir. The air quality measurements in the boning hall showed a gradual build-up of carbon dioxide and aerosol particles over the course of a work shift, confirming that this poorly ventilated area of the plant had an environment that was highly favorable for aerosol transmission of SARS-CoV-2. Assessment of operational conditions incorporated visual surveys of the plant during the working day. Prior to and during the first 2 weeks of the outbreak, multiple measures were introduced into the plant by management, including physical distancing, provision of educational material to workers, visitor restrictions, and environmental monitoring. After the implementation of these measures and their progressive refinement by plant management, the factory had no further linked cases (clusters) or outbreaks for the following 198 days. The tailored approach to risk mitigation adopted in this meat processing plant shows that generic risk mitigation measures, as recommended by public health authorities, can be successfully adapted and optimized by designated plant emergency response teams.

A Multiomic Approach to Investigate the Effects of a Weight Loss Program on the Intestinal Health of Overweight Horses.

Obesity is endemic in human populations in the western society, and with mounting evidence that the intestinal ecological environment plays a major role in its pathogenesis, identification of therapies based on intestinal microbiota modulation are gaining attention. Obesity in companion animals is also a common clinical problem. We set out using a multidimensional approach, to determine the effectiveness and safety of a weight loss program for horses incorporating diet restriction and exercise. In addition, we aimed to investigate the effect of this program on the overall intestinal health of overweight sedentary horses. The investigation comprised of a randomized, controlled, 6-week study of 14 overweight sedentary horses and ponies who were blocked for age, gender, and breed (controls n = 7, treatment n = 7). The treatment group were fed a restricted diet (1.4% of body weight dry matter intake) and the control group a maintenance diet (2% of body weight as dry matter intake) over the study period. The treatment group were subjected to a prescribed exercise regime, while the control group were exercised to mimic foraging conditions. Several clinical measurements were taken at the start and end of the study, including morphological parameters, ultrasound measurements of subcutaneous fat, and blood pressure. Fecal microbiota analysis was performed using 16S rRNA gene sequence analysis, and fecal metabolome was analyzed using NMR spectroscopy, on samples taken at weeks 1, 3, and 6 of the study. All horses completed the study period successfully. However, two of the treatment group had to have modified exercise regimes. The treatment group showed significant weight loss (p < 0.00001) and an associated decrease in waste circumference (p < 0.0001) when compared with the control group. The alpha-diversity of the fecal microbiota in the treatment group showed a significant increase from the start to the end of the study period (p < 0.05); however, there was no significant difference between groups at any sampling point. There were significant changes (p < 0.05) in the metabolome in both groups between the start and end of the study, but not between groups at any sampling point. Finally, the resting blood pressure of all horses was significantly lower by the end of the study.

Making Nature Explicit in Children's Drawings of Wellbeing and Happy Spaces.

Previous research on children's wellbeing indicators has focused extensively on adults', rather than children's perspectives, despite there being a broad consensus that children's conceptualisations differ significantly from adults'. In response, this study aimed to explore what constitutes children's wellbeing through their drawings and discussions. Ninety-one seven and eight-year old children from two primary schools in areas of relatively high deprivation in eastern England participated in this study. We identified indicators of wellbeing that were made explicit in children's drawings, such as the need for safety, happiness and positive relationships, but also indicators that remained rather implicit, such as the environment and nature. The drawings in particular illustrated that children's perceptions of wellbeing were subject to the affordances of their favourite spaces for emotional, mental physical and material wellbeing. Access to nature and outdoor spaces was interconnected with all these affordances. We analysed these findings through the theoretical lenses of positive psychology, self-actualisation, social mentality and the human-nature relationship. We argue that making nature explicit, and restoring the interconnectedness between the arts and nature in the current literature, should be a key priority for future research and practice on children's wellbeing indicators.

Outbreak of acute larval cyathostominosis - A "perfect storm" of inflammation and dysbiosis.

BACKGROUND: Cyathostomins are prevalent and pathogenic intestinal helminths of horses, causing acute and chronic disease, including acute larval cyathostominosis, which has a mortality rate of 50%. Factors determining individual susceptibility to acute larval cyathostominosis are unknown. Investigation of these factors could lead to novel treatment and prevention strategies. OBJECTIVES: To investigate clinicopathological and faecal microbiota changes associated with disease in individual horses in an acute larval cyathostominosis outbreak. STUDY DESIGN: Case series. METHODS: The study population was a herd of 23 mixed breed horses in Ireland. The outbreak occurred in November 2018. Fourteen horses were clinically affected. Clinical status was monitored and recorded. Blood and faecal sampling allowed clinicopathological, faecal 16s rRNA gene sequencing and faecal egg count analyses. RESULTS: Two horses were euthanised, whilst 12 recovered. Common clinical signs included loose faecal consistency, weight loss and pyrexia. Consistent clinicopathological findings were borderline anaemia, leucocytosis, thrombocytosis, hyperfibrinogenaemia, hyperglobulinaemia and a reverse A: G ratio. Decreased alpha-diversity of the faecal microbiota and greater relative abundance of the genus Streptococcus, class Bacilli, order Lactobacillales and family Streptococcaceae, and family Prevotelleceae was found in clinically affected horses compared to their clinically normal cohorts. An increase in obligate fibrolytic bacteria was seen in the clinically normal group compared to the clinical group. Histopathological findings of the colon and caecum revealed a severe necrotising typhlocolitis associated with cyathostomin larvae and bacterial overgrowth in the mucosa of the large intestine. MAIN LIMITATIONS: The study population in this outbreak is small. There are several confounding factors limiting this to a descriptive case series. Faecal microbiota has been shown to reflect the large intestinal microbiota but do not represent changes directly. CONCLUSIONS: These findings suggest that acute larval cyathostominosis is associated with dysbiosis of the gut microbiota as well as the inflammatory stimulus of numerous emerging larvae leading to structural and functional pathology of the large intestine.

No Worm Is an Island; The Influence of Commensal Gut Microbiota on Cyathostomin Infections.

The importance of the gut microbiome for host health has been the subject of intense research over the last decade. In particular, there is overwhelming evidence for the influence of resident microbiota on gut mucosal and systemic immunity; with significant implications for the outcome of gastrointestinal (GI) infections, such as parasitic helminths. The horse is a species that relies heavily on its gut microbiota for GI and overall health, and disturbances in this complex ecosystem are often associated with life-threatening disease. In turn, nearly all horses harbour parasitic helminths from a young age, the most prevalent of which are the small strongyles, or cyathostomins. Research describing the relationship between gut microbiota and cyathostomin infection is in its infancy, however, to date there is evidence of meaningful interactions between these two groups of organisms which not only influence the outcome of cyathostomin infection but have long term consequences for equine host health. Here, we describe these interactions alongside supportive evidence from other species and suggest novel theories and avenues for research which have the potential to revolutionize our approach to cyathostomin prevention and control in the future.

Removal of adult cyathostomins alters faecal microbiota and promotes an inflammatory phenotype in horses.

The interactions between parasitic helminths and gut microbiota are considered to be an important, although as yet incompletely understood, factor in the regulation of immunity, inflammation and a range of diseases. Infection with intestinal helminths is ubiquitous in grazing horses, with cyathostomins (about 50 species of which are recorded) predominating. Consequences of infection include both chronic effects, and an acute inflammatory syndrome, acute larval cyathostominosis, which sometimes follows removal of adult helminths by administration of anthelmintic drugs. The presence of cyathostomins as a resident helminth population of the equine gut (the "helminthome") provides an opportunity to investigate the effect helminth infection, and its perturbation, has on both the immune system and bacterial microbiome of the gut, as well as to determine the specific mechanisms of pathophysiology involved in equine acute larval cyathostominosis. We studied changes in the faecal microbiota of two groups of horses following treatment with anthelmintics (fenbendazole or moxidectin). We found decreases in both alpha diversity and beta diversity of the faecal microbiota at Day 7 post-treatment, which were reversed by Day 14. These changes were accompanied by increases in inflammatory biomarkers. The general pattern of faecal microbiota detected was similar to that seen in the relatively few equine gut microbiome studies reported to date. We conclude that interplay between resident cyathostomin populations and the bacterial microbiota of the equine large intestine is important in maintaining homeostasis and that disturbance of this ecology can lead to gut dysbiosis and play a role in the aetiology of inflammatory conditions in the horse, including acute larval cyathostominosis.