Horses vaccinated with live attenuated intranasal strangles vaccine seroconvert to SEQ2190 and SeM.

BACKGROUND: The dual antigen iELISA uses two Streptococcus equi subsp equi surface protein antigens composed of N-terminal portions of SEQ2190 (Antigen A) and SeM (Antigen C). It is currently used to identify animals exposed to S. equi which have developed an immune response to the target antigens. OBJECTIVES: To determine the usefulness of the dual antigen iELISA in a population of horses vaccinated with Pinnacle IN. We hypothesised that horses vaccinated for strangles with a live attenuated, non-encapsulated SeM-2 strain of S. equi, would seroconvert when tested 5 weeks later by the dual antigen iELISA. STUDY DESIGN: Prospective case-control study. METHODS: Three separate serum samples were obtained from 26 client-owned horses vaccinated annually with Pinnacle® IN and 26 university-owned (non-vaccinates): at annual strangles vaccination (S1), 5-week post-vaccination (S2) from vaccinates, and a third (S3) (at 10 weeks) from vaccinates who received a booster. Seropositivity was defined as an OD450 nm value ≥0.5 for one or both antigens. Mixed-effects ordered logistic regression analysis was used to identify factors associated with a suspect seropositive and seropositive value on the combined Antigen A and Antigen C iELISA. Post hoc pairwise comparisons of linear predictive margins were used to assess the differences in OD450 at a specific time between Antigens A and C. RESULTS: Nineteen of 25 (76%) vaccinates were seropositive at S2 compared to 1 of 26 (4%) non-vaccinates. When adjusted for sample number, vaccinates were more likely to be seropositive or suspect than non-vaccinates (OR 14; P = .02, 95% CI 1.62-122.03). The OD450 value was significantly larger for Antigen C than Antigen A for vaccinates (P < .001; 95% CI 0.13-0.26) when normalised by age, sex and breed. MAIN LIMITATIONS: Guttural pouch sampling for S. equi in seroconverted horses was unavailable. CONCLUSIONS: With a high rate of seroconversion to both antigens, the use of the dual antigen iELISA is not recommended in populations vaccinated with Pinnacle® IN.

Globetrotting strangles: the unbridled national and international transmission of Streptococcus equi between horses.

The equine disease strangles, which is characterized by the formation of abscesses in the lymph nodes of the head and neck, is one of the most frequently diagnosed infectious diseases of horses around the world. The causal agent, Streptococcus equi subspecies equi, establishes a persistent infection in approximately 10 % of animals that recover from the acute disease. Such 'carrier' animals appear healthy and are rarely identified during routine veterinary examinations pre-purchase or transit, but can transmit S. equi to naïve animals initiating new episodes of disease. Here, we report the analysis and visualization of phylogenomic and epidemiological data for 670 isolates of S. equi recovered from 19 different countries using a new core-genome multilocus sequence typing (cgMLST) web bioresource. Genetic relationships among all 670 S. equi isolates were determined at high resolution, revealing national and international transmission events that drive this endemic disease in horse populations throughout the world. Our data argue for the recognition of the international importance of strangles by the Office International des Épizooties to highlight the health, welfare and economic cost of this disease. The Pathogenwatch cgMLST web bioresource described herein is available for tailored genomic analysis of populations of S. equi and its close relative S. equi subspecies zooepidemicus that are recovered from horses and other animals, including humans, throughout the world. This article contains data hosted by Microreact.

Seroprevalence of Streptococcus equi subspecies equi in Croatia - Short communication.

Clinical cases resembling strangles are regularly seen in some areas of Croatia. However, there are no data on the prevalence of infection and the clinical forms or geographic distribution of the disease. The aim of this study was to determine the seroprevalence of Streptococcus equi subspecies equi in horses resident in Croatia, in order to estimate the geographic distribution of infection. The study included 291 horse sera from the eight counties where the majority of Croatian horses are kept. Sera were tested by indirect ELISA (iELISA) for the presence of serum antibodies against S. equi protein A (SEQ_2190) and protein C (SeM). Positive horses were detected in all counties. Overall seroprevalence was 16.5 per cent (48/291), ranging from 7.1 to 29.6 per cent. A positive association was observed between the population size of the horses in the counties and the seropositivity rates: the larger the population, the higher the seropositivity. The results of this study suggest that S. equi infection is widespread in Croatia. Further investigation of the clinical manifestations, circulating strains and other characteristics of the disease in Croatia and raising awareness of the disease among horse owners are now required.

Identification of genes required for the fitness of Streptococcus equi subsp. equi in whole equine blood and hydrogen peroxide.

The availability of next-generation sequencing techniques provides an unprecedented opportunity for the assignment of gene function. Streptococcus equi subspecies equi is the causative agent of strangles in horses, one of the most prevalent and important diseases of equids worldwide. However, the live attenuated vaccines that are utilized to control this disease cause adverse reactions in some animals. Here, we employ transposon-directed insertion-site sequencing (TraDIS) to identify genes that are required for the fitness of S. equi in whole equine blood or in the presence of H2O2 to model selective pressures exerted by the equine immune response during infection. We report the fitness values of 1503 and 1471 genes, representing 94.5 and 92.5 % of non-essential genes in S. equi, following incubation in whole blood and in the presence of H2O2, respectively. Of these genes, 36 and 15 were identified as being important to the fitness of S. equi in whole blood or H2O2, respectively, with 14 genes being important in both conditions. Allelic replacement mutants were generated to validate the fitness results. Our data identify genes that are important for S. equi to resist aspects of the immune response in vitro, which can be exploited for the development of safer live attenuated vaccines to prevent strangles.

Streptococcus canis multilocus sequence typing in a case series of dogs with ulcerative keratitis.

OBJECTIVE: To determine whether four isolates of Streptococcus canis (S canis) recovered from dogs diagnosed with ulcerative keratitis at the Animal Health Trust (AHT) were genetically related to other ocular isolates that are registered in the online database. ANIMAL STUDIED: Four S canis corneal isolates. PROCEDURES: Clinical and laboratory records between 2016 and 2017 were searched for dogs with ulcerative keratitis for which microbiology analysis was consistent with the growth of S canis. Genomic DNA was extracted for sequencing (Illumina MiSeq), and multilocus sequence types (STs) were determined using MLST 1.8 relative to the 44 sequence types of S canis available. A neighbor-joining tree was constructed in MEGA v4.0. A two-sided Fisher's exact test was used to determine any associations between the isolated strains and ocular infections of dogs. RESULTS: Four strains were isolated from pugs (cases 1-4) with ulcerative keratitis. Genome sequencing identified ST-27 (case 1), ST-9 (case 3), and ST-13 (cases 2 and 4). STs 13 and 27 are members of Clonal Complex (CC)-13. Analysis of the multilocus sequence typing database revealed that CC-13 strains accounted for six of the twelve isolates recovered from the eye exudates of dogs (P = .0078). CONCLUSIONS: There is early evidence that the CC-13 group of S canis is associated with ocular infections in dogs. We provide draft genome sequences toward the future identification of virulence mechanisms associated with streptococcal keratitis in dogs.

Using next-generation sequencing to determine diversity of horse intestinal worms: identifying the equine 'nemabiome'.

Next-generation sequencing of DNA from nematode eggs has been utilised to give the first account of the equine 'nemabiome'. In all equine faecal samples investigated, multiple species of Strongylidae were detected, ranging from 7.5 (SEM 0.79) with 99+% identity to sequences in the NCBI database to 13.3 (SEM 0.80) with 90+% identity. This range is typical of the number of species described previously in morphological studies using large quantities of digesta per animal. However, the current method is non-invasive; relies on DNA analysis, avoiding the need for specialist microscopy identification; and can be carried out with small samples, providing significant advantages over current methods.

Timelines in the diagnostic evaluation of people with suspected amyotrophic lateral sclerosis (ALS)/motor neuron disease (MND)--a 20-year review: can we do better?

This paper examines diagnostic timelines for people suspected of having ALS/MND over a 20-year period, evaluates the impact of a 'fast track' diagnostic process and discusses typical causes of diagnostic delay. Key diagnostic timelines (dates of first symptom, diagnosis and death) were reviewed for people diagnosed between 1989 and 2008. Patients evaluated through a fast-track diagnostic process and those investigated through traditional neurology clinics were compared. Typical causes of diagnostic delay were investigated. Results showed that diagnostic timelines have been surprisingly consistent over this 20-year period. Time from first symptom to diagnosis hovered around 12 months, with the diagnosis typically being made around the midpoint (50% of total disease duration elapsed) of the disease pathway. The introduction of a fast-track process has not to date affected overall performance but has shortened times from referral to diagnosis. Diagnostic delays appear to be associated with clinical complexity and delays in referral, both within primary and secondary care services. In conclusion, more widespread implementation of fast-track processes could potentially reduce diagnostic delays. Educational interventions among health care professionals both in primary and secondary care may also help shorten diagnostic pathways.