A risk assessment of equine piroplasmosis entry, exposure and consequences in the UK.

BACKGROUND: Equine piroplasmosis (EP) is currently not endemic in the UK, despite a lack of formal surveillance and the presence of carrier horses in the equine population. Pathogen establishment would have significant welfare and economic impacts on the national equine industry, but the disease is often overlooked by UK practitioners. OBJECTIVES: To assess the risk of disease entry, exposure and consequences to the UK equine population. STUDY DESIGN: Qualitative risk assessment. METHODS: A qualitative risk assessment was constructed utilising the current World Organisation for Animal Health (OIE) published framework for importation risk assessment, assessing the key areas of disease entry, exposure and consequences to the UK equine population. RESULTS: The overall risk of EP entry to the UK via importation of infected equidae with acute disease is very low but considered medium with subclinical carrier animals. Entry via importation of ticks or the importation of blood is considered very low. The risk of EP exposure to susceptible equidae in the UK is considered low by the infection routes of tick-bites, contaminated needles and contaminated blood, but very high via transplacental transfer. However, the consequences of EP endemic establishment are considered of high significance to the UK equine industry. MAIN LIMITATIONS: A lack of available numerical data for events and variables in disease import risk meant a qualitative assessment was the most practical method for this scenario. CONCLUSIONS: This risk assessment highlights that EP positive animals are able to enter and are currently present in the UK, and that conditions do exist that could allow forward transmission of the disease. It has highlighted a gap in existing policy where the UK falls behind OIE guidelines and has suggested steps to correct this discrepancy and improve national biosecurity.

A study comparing the healthy and diseased equine glandular gastric microbiota sampled with sheathed transendoscopic cytology brushes.

The role of the equine gastrointestinal microbiota in the pathogenesis of equine glandular gastric disease (EGGD) is poorly understood. To investigate whether the glandular gastric microbiota is altered in horses with EGGD. Prospective longitudinal study. Five Thoroughbred racehorses from one training center underwent gastroscopy as part of poor performance investigation. Samples were taken from EGGD lesions and adjacent normal mucosa using sheathed transendoscopic cytology brushes and frozen at -80°C. DNA was extracted for 16S rRNA sequencing, and sequences compared against a database to generate taxonomic classification of the microbiota. The same horses were sampled 6 months later. Normal glandular mucosal samples were characterized by a higher proportion of Proteobacteria (46.3%) than EGGD lesions (18.9%). Relative abundance of Firmicutes was lower in samples from normal mucosa (20.0%) than EGGD lesions (41.2%). Linear discriminant analysis effect size (LEfSe) confirmed a greater proportion of Firmicutes species was characteristic of samples collected from EGGD lesions due to a very high relative abundance of Sarcina (up to 92.4%) in two horses with EGGD. We were unable to comment on the stability of the glandular gastric microbiota over time. Small sample population. None of the horses examined had grossly normal gastric mucosa. The gastric microbiota appears altered in EGGD, although we are unable to demonstrate a causative effect. Sarcina was particularly increased in abundance in EGGD and may be a useful biomarker of disease. Sheathed cytology brushes were an effective method for sampling the gastric mucosa.

Comparison of loop-mediated isothermal amplification (LAMP) and PCR for the diagnosis of infection with Trypanosoma brucei ssp. in equids in The Gambia.

INTRODUCTION: Infection of equids with Trypanosoma brucei (T. brucei) ssp. is of socioeconomic importance across sub-Saharan Africa as the disease often progresses to cause fatal meningoencephalitis. Loop-mediated isothermal amplification (LAMP) has been developed as a cost-effective molecular diagnostic test and is potentially applicable for use in field-based laboratories. PART I: Threshold levels for T. brucei ssp. detection by LAMP were determined using whole equine blood specimens spiked with known concentrations of parasites. Results were compared to OIE antemortem gold standard of T. brucei-PCR (TBR-PCR). RESULTS I: Threshold for detection of T. brucei ssp. on extracted DNA from whole blood was 1 parasite/ml blood for LAMP and TBR-PCR, and there was excellent agreement (14/15) between tests at high (1 x 103/ml) concentrations of parasites. Detection threshold was 100 parasites/ml using LAMP on whole blood (LWB). Threshold for LWB improved to 10 parasites/ml with detergent included. Performance was excellent for LAMP at high (1 x 103/ml) concentrations of parasites (15/15, 100%) but was variable at lower concentrations. Agreement between tests was weak to moderate, with the highest for TBR-PCR and LAMP on DNA extracted from whole blood (Cohen's kappa 0.95, 95% CI 0.64-1.00). PART II: A prospective cross-sectional study of working equids meeting clinical criteria for trypanosomiasis was undertaken in The Gambia. LAMP was evaluated against subsequent TBR-PCR. RESULTS II: Whole blood samples from 321 equids in The Gambia were processed under field conditions. There was weak agreement between LWB and TBR-PCR (Cohen's kappa 0.34, 95% CI 0.19-0.49) but excellent agreement when testing CSF (100% agreement on 6 samples). CONCLUSIONS: Findings support that LAMP is comparable to PCR when used on CSF samples in the field, an important tool for clinical decision making. Results suggest repeatability is low in animals with low parasitaemia. Negative samples should be interpreted in the context of clinical presentation.

Variation in disease phenotype is marked in equine trypanosomiasis.

BACKGROUND: Equine trypanosomiasis is a severe and prevalent disease that has the greatest impact globally upon working equids due to its distribution across lower income countries. Morbidity and mortality rates are high; disease management strategies in endemic regions are ineffective and cost prohibitive. Individual variation in disease phenotype in other species suggests host factors could reveal novel treatment and control targets but has not been investigated in equids. METHODS: A prospective clinical evaluation of equines presenting for a free veterinary examination was performed in hyperendemic villages in The Gambia. Age, body condition score and body weight were estimated by validated methods, and haematocrit and total protein concentration measured. Animals fulfilling 2 out of 5 clinical inclusion criteria (anaemia, poor body condition, pyrexia, history of abortion, oedema) for a diagnosis of trypanosomiasis received trypanocidal treatment with follow-up at 1 and 2 weeks. Blood samples underwent PCR analysis with specific Trypanosoma spp. primers and results were compared to the subject's clinical and clinicopathological features. A mixed effects generalised linear model was generated to evaluate the association of infection status with degree of pyrexia and anaemia. RESULTS: Morbidity was high within examined (n = 641) and selected (n = 247) study populations. PCR status was not associated with a defined disease phenotype; there was intra- and inter-species variability. Donkeys were more frequently Trypanosoma spp.-positive (P < 0.001) and febrile (P < 0.001) than horses, but infected horses were more anaemic (P < 0.001), and in poorer body condition (P < 0.001) than donkeys. Sex was correlated to disease phenotype: males were more anaemic (P = 0.03) and febrile (P < 0.001). Haemoparasite co-infections were more common than a single infection. CONCLUSIONS: There was evidence of diversity in trypanosomiasis clinical signs plus variable disease phenotypes within equid subpopulations that warrant further investigation. The complex co-infection profile of field cases requires greater consideration to optimise disease management.

Analysis of Theileria equi diversity in The Gambia using a novel genotyping method.

Theileria equi, one of the primary pathogens causing equine piroplasmosis, has previously been sub-classified into a number of clades on the basis of 18S SSU rRNA gene sequence diversity. This partitioning of the parasite population has potential implications for host immunity, treatment and vaccine development. To detect and identify different clade genotypes among and within individual equine blood samples, a novel PCR-based technique was designed and optimized. Theileria equi has only recently been described in The Gambia, and the developed genotyping technique was used to analyse blood samples taken from 42 piroplasmosis-positive horses and donkeys within the country. Three different T. equi genotypes were detected within the population, including the same genotype as the recently described Theileria haneyi, with 61.9% of individuals found to be infected with more than one genotype. Overall, there was a trend that males were more likely to have a multiple genotype infection. Thus, the novel genotyping technique has been shown to be effective in analysis of field populations and offers researchers a rapid method of identifying multiple T. equi genotypes both within individuals and equine populations in epidemiological studies.

Safety and efficacy of three trypanocides in confirmed field cases of trypanosomiasis in working equines in The Gambia: a prospective, randomised, non-inferiority trial.

BACKGROUND: Globally, working equines have a continued and growing socioeconomic role in supporting the livelihoods of between 300-600 million people in low income countries which is rarely recognised at a national or international level. Infectious diseases have significant impact on welfare and productivity in this population and equine trypanosomiasis is a priority disease due to its severity and prevalence. Strategies are required to improve the prevention, diagnosis, management and treatment of trypanosomiasis in equines and more data are required on the efficacy and safety of current trypanocidal drugs. METHODS: A prospective randomised, open-label non-inferiority trial was performed in The Gambia on horses and donkeys that fulfilled 2/5 clinical inclusion criteria (anaemia, poor body condition, pyrexia, history of abortion, oedema). Following randomised trypanocidal treatment (diminazene diaceturate, melarsomine dihydrochloride or isometamidium chloride), animals were observed for immediate adverse drug reactions and follow-up assessment was performed at 1 and 2 weeks. Blood samples underwent PCR analysis with specific Trypanosoma sp. primers. Treatment efficacy was assessed by measuring changes in clinical parameters, clinicopathological results and PCR-status post-treatment after evaluating for bias. Using PCR status as the outcome variable, non-inferiority of isometamidium treatment was determined if the upper bound limit of a 2-sided 95% CI was less than 10%. RESULTS: There was a significant beneficial effect upon the Trypanosoma sp. PCR positive population following trypanocidal treatment for all groups. The findings of clinical evaluation and PCR status supported a superior treatment effect for isometamidium. Melarsomine dihydrochloride efficacy was inferior to isometamidium. There were immediate, self-limiting side effects to isometamidium in donkeys (26%). Diminazene had the longest duration of action as judged by PCR status. CONCLUSIONS: The data support the continued use of isometamidium following careful dose titration in donkeys and diminazene for trypanosomiasis in equines using the doses and routes of administration reported.

VALIDATION OF A BEAMNRC MONTE CARLO SIMULATION OF A BROAD BEAM DIAGNOSTIC X-RAY UNIT.

A BEAMnrc Monte Carlo simulation of a diagnostic X-ray unit in a broad beam geometry intended for the derivation of effective linear attenuation coefficients was validated against measurements made on the X-ray unit on which the simulation was based. The validation process assessed the size of the beam, the first and second half value layer, the variation in kerma with anode-heel effect and the accuracy of values of effective linear attenuation coefficient for water and solid water attenuators. The agreement between the simulated and measured results for all tests was consistently within the experimental uncertainty for the measurements. The average deviation between values of effective linear attenuation coefficient for simulated and measured results is 3.8%, with the highest individual deviation 7.1%. The simulation can be used to produce values of effective linear attenuation coefficient for a range of kVp, field size and attenuator thickness that are close to those measured.

Equine piroplasmosis status in the UK: an assessment of laboratory diagnostic submissions and techniques.

Equine piroplasmosis (EP) has historically been of minor concern to UK equine practitioners, primarily due to a lack of competent tick vectors. However, increased detection of EP tick vector species in the UK has been reported recently. EP screening is not currently required for equine importation, and when combined with recent relaxations in movement regulations, there is an increased risk regarding disease incursion and establishment into the UK. This study evaluated the prevalence of EP by both serology and PCR among 1242 UK equine samples submitted for EP screening between February and December 2016 to the Animal and Plant Health Agency and the Animal Health Trust. Where information was available, 81.5 per cent of submissions were for the purpose of UK export testing, and less than 0.1 per cent for UK importation. Serological prevalence of EP was 8.0 per cent, and parasite DNA was found in 0.8 per cent of samples. A subsequent analysis of PCR sensitivity in archived clinical samples indicated that the proportion of PCR-positive animals is likely to be considerably higher. The authors conclude that the current threat imposed by UK carrier horses is not adequately monitored and further measures are required to improve national biosecurity and prevent endemic disease.

Determining paediatric patient thickness from a single digital radiograph-a proof of principle.

OBJECTIVE: This work presents a proof of principle for a method of estimating the thickness of an attenuator from a single radiograph using the image, the exposure factors with which it was acquired and a priori knowledge of the characteristics of the X-ray unit and detector used for the exposure. It is intended this could be developed into a clinical tool to assist with paediatric patient dose audit, for which a measurement of patient size is required. METHODS: The proof of principle used measured pixel value and effective linear attenuation coefficient to estimate the thickness of a Solid Water attenuator. The kerma at the detector was estimated using a measurement of pixel value on the image and measured detector calibrations. The initial kerma was estimated using a lookup table of measured output values. The effective linear attenuation coefficient was measured for Solid Water at varying kVp. 11 test images of known and varying thicknesses of Solid Water were acquired at 60, 70 and 81 kVp. Estimates of attenuator thickness were made using the model and the results compared to the known thickness. RESULTS: Estimates of attenuator thickness made using the model differed from the known thickness by 3.8 mm (3.2%) on average, with a range of 0.5-10.8 mm (0.5-9%). CONCLUSION: A proof of principle is presented for a method of estimating the thickness of an attenuator using a single radiograph of the attenuator. The method has been shown to be accurate using a Solid Water attenuator, with a maximum difference between estimated and known attenuator thickness of 10.8 mm (9%). The method shows promise as a clinical tool for estimating abdominal paediatric patient thickness for paediatric patient dose audit, and is only contingent on the type of data routinely collected by Medical Physics departments. Advances in knowledge: A computational model has been created that is capable of accurately estimating the thickness of a uniform attenuator using only the radiographic image, the exposure factors with which it was acquired and a priori knowledge of the characteristics of the X-ray unit and detector used for the exposure.

Erratum to: Arrhythmogenic right ventricular cardiomyopathy secondary to adipose infiltration as a cause of episodic collapse in a horse.

[This corrects the article DOI: 10.1186/s13620-015-0052-3.].

Molecular confirmation of Sarcocystis fayeri in a donkey.

Sarcocystis fayeri is a canine protozoan parasite with an equine intermediate host. Historically classified as an incidental pathogen, recent literature has described the toxic effects of Sarcocystis fayeri in human food poisoning, and highlighted potential involvement in equine neuromuscular disease. Until now, horses were believed to be the exclusive intermediate host. This study reports the first molecular confirmation of S. fayeri in a donkey, and gives rise to the consideration of donkeys being a potential reservoir for the parasite. This finding is of particular importance in understanding the epidemiology of this disease.

The application of exhaled breath analysis in racing Thoroughbreds and the influence of high intensity exercise and ambient temperature on the concentration of carbon monoxide and pH in exhaled breath.

Analyses of exhaled breath (EB) and exhaled breath condensate (EBC) are non-invasive modalities for assessing the lower airways but these methods have not been applied to Thoroughbred racehorses in training. The aims of this study were to determine whether EB and EBC could be obtained from Thoroughbred racehorses in the field and to investigate the effects of exercise per se and during different ambient temperatures and humidity on exhaled concentrations of nitric oxide (eNO), carbon monoxide (eCO) and EBC pH. EB and EBC samples were obtained from 28 Thoroughbred racehorses pre- and post-exercise during warm (n=23) and/or cold (n=19) ambient temperatures. eNO was detected in 19/84 EB samples. eCO was measured in 39/42 EB samples pre-exercise (median 1.3 ppm) and concentrations decreased significantly post-exercise (median 0.8 ppm, P<0.005) and were associated with ambient temperature. EBC pH was 4.51 ± 0.23 pre-exercise and increased significantly post-exercise (4.79 ± 0.59, P=0.003). The study documented the collection of EB and EBC from Thoroughbred racehorses in a field setting. Alterations in concentrations of volatile gases and EBC pH occurred in response to exercise, and were likely to have been influenced by environmental factors.

Functional intercomparison of intraoperative radiotherapy equipment - Photon Radiosurgery System.

BACKGROUND: Intraoperative Radiotherapy (IORT) is a method by which a critical radiation dose is delivered to the tumour bed immediately after surgical excision. It is being investigated whether a single high dose of radiation will impart the same clinical benefit as a standard course of external beam therapy. Our centre has four Photon Radiosurgery Systems (PRS) currently used to irradiate breast and neurological sites. MATERIALS AND METHODS: The PRS comprises an x-ray generator, control console, quality assurance tools and a mobile gantry. We investigated the dosimetric characteristics of each source and its performance stability over a period of time. We investigated half value layer, output diminution factor, internal radiation monitor (IRM) reproducibility and depth-doses in water. The half value layer was determined in air by the broad beam method, using high purity aluminium attenuators. To quantify beam hardening at clinical depths, solid water attenuators of 5 and 10 mm were placed between the x-ray probe and attenuators. The ion chamber current was monitored over 30 minutes to deduce an output diminution factor. IRM reproducibility was investigated under various exposures. Depth-dose curves in water were obtained at distances up to 35 mm from the probe. RESULTS: The mean energies for the beam attenuated by 5 and 10 mm of solid water were derived from ICRU Report 17 and found to be 18 and 24 keV. The average output level over a period of 30 minutes was found to be 99.12%. The average difference between the preset IRM limit and the total IRM count was less than 0.5%. For three x-ray sources, the average difference between the calculated and actual treatment times was found to be 0.62% (n = 30). The beam attenuation in water varied by approximately 1/r(3). CONCLUSION: The x-ray sources are stable over time. Most measurements were found to lie within the manufacturer's tolerances and an intercomparison of these checks suggests that the four x-ray sources have similar performance characteristics.