Sequelae and complications of Streptococcus equi subspecies equi infections in the horse

Streptococcus equi ssp. equi infection in the horse, or strangles, commonly results in abscessation of the submandibular, submaxillary or retropharyngeal lymph nodes. Although this classical presentation of strangles is associated with a low mortality rate, complications and sequlae may worsen the prognosis and increase mortality rates. This article reviews sequelae and complictions of S. equi ssp. equi infection in the horse, including guttural pouch empyema, bastard strangles and immune mediated diseases such as purpura haemorrhagica, myopathies and myocarditis.     

Molecular characterisation of 'strangles' outbreaks in the UK: the use of M-protein typing of Streptococcus equi ssp. equi

Reasons for performing study: Strangles is the most commonly diagnosed and important infectious disease of horses worldwide. Very little is known about the temporo‐spatial and molecular epidemiology of strangles. The disease is not notifiable in the UK and there are few published data on the geographical locations of outbreaks. Objective: To investigate whether typing of a surface protein (SeM) of Streptococcus equi ssp. equi (S. equi), the causative agent of strangles, is a useful epidemiological tool. Methods: The variable region of the SeM gene was amplified from 145 isolates of S. equi by PCR and sequenced. Different SeM gene alleles were assigned based on the SeM database, grouped into phylogenetic clusters using split decomposition analysis and plotted against the submitting veterinary practices. Results: In this study 21 S. equi SeM alleles were found, including 9 previously unidentified alleles and representing 4 phylogenetic groups. S. equi containing SeM alleles 9 and 7 were the most commonly isolated and there was a high number of low frequency alleles. The occurrence of an outbreak cluster in the north‐west of the UK is also reported. Conclusions: Strangles outbreaks can be differentiated on the basis of their SeM allele sequences. The data provide further evidence of SeM mutation leading to the emergence of novel, but related SeM alleles that are geographically linked. Sequencing of the SeM gene is a useful tool for the elucidation of strangles epidemiology at a regional and a national level. Potential relevance: This technique may allow differentiation or linkage of strangles outbreaks and as such may be an effective tool for local as well as national and international disease surveillance.     

Changes in Serum Antibody Levels after Vaccination for Strangles and after Intranasal Challenge with Streptococcus equi subsp. equi in Horses

In this study, to evaluate the influence of strangles vaccination on serological test results, we investigated the changes in strangles serum antibody levels in horses after vaccination and subsequent intranasal challenge with S. equi. The horses were vaccinated for strangles with either a component vaccine (Group C) or a live vaccine (Group L). We measured changes in strangles serum antibody levels weekly for 20 weeks after vaccinating horses twice for strangles over a 3-week interval, and for 7 weeks after intranasal challenge with S. equi in the same horses. Serum antibody responses to the proline-glutamic acid-proline-lysine (PEPK) antigen with five repetitions (PEPK-5R) were higher at all times (up to 2.4-fold) following vaccination in Group C than in Group L, and the value peaked at 2.9-fold above the initial value after the second vaccination in Group C horses. However, the value was lower than that in horses infected with S. equi, and it gradually decreased, reaching the initial (week 0) value by the 15th week. Serum antibody responses to PEPK-5R after challenge with S. equi increased in both groups of horses, but the value tended to be lower than that reported for unvaccinated horses. In addition, the average value in Group C was 2.6-fold higher than that of Group L. These results suggest the serum antibody responses of horses infected with S. equi varies according to the type of vaccine with which they have been vaccinated. Although the serological diagnostic test for strangles in which PEPK-5R is used as an antigen is effective for the investigation of serum antibodies to strangles in vaccinated horses, the present data suggest it is necessary to consider the vaccination history when interpreting the results.     

Subscapular lymph node abscessation as a result of metastatic Streptococcus equi subspecies equi infection: An atypical presentation of bastard strangles in a mare

The case reported here represents an atypical presentation of bastard strangles in an 18‐year‐old Arab mare. The horse initially presented for progressive neck pain characterised by reluctance to lateral and ventroflexion of the neck. Subsequent diagnostics revealed a subscapular abscess and aspirates of the mass cultured positive for Streptococcus equi ssp. equi. Surgical drainage and debridement of the abscess was performed under general anaesthesia. Six months post surgery, the mare had made a complete recovery.     

First Report of Molecular Characterization of Argentine Isolates of Streptococcus equi subsp. equi by Pulsed-Field Gel Electrophoresis

Strangles is one of the most frequently diagnosed equine respiratory infectious diseases in the world. It is caused by Streptococcus equi subsp. equi (S. equi), and it is an acute infection characterized by pyrexia, nasal discharge, pharyngitis, and abscessation of lymph nodes. Frequently, healthy horses might continue to harbor S. equi after clinical recovery. Although the genetic distance between S. equi isolates is short, strains can be differentiated by pulsed-field gel electrophoresis (PFGE) and single locus sequence typing for epidemiological studies. The aim of this study was to characterize by PFGE Argentine isolates of S. equi obtained from horses with acute strangles and those that had recovered. Bacterial isolation and identification of 80 S. equi isolates by phenotypic and genotypic tests were performed using samples from 29 horses with acute strangles and 95 from healthy animals. Also, the isolates were characterized by PFGE using Bsp120I and SmaI. Visual comparison of macrorestriction patterns generated with both enzymes revealed three different DNA fragment profiles with variations of one or two bands. Interestingly, an identical profile was found in isolates from the same horse and from horses that were infected at the same time, and the horses recovered from strangles continue to carry the same strain. Some vaccinated horses have been mild infected for a different strain from that of carriers suggesting other source of infection. This is the first molecular characterization of Argentine isolates of S. equi, which shows the presence of three strains between 2010 and 2013 in Buenos Aires.     

Prevalence and serum protein values of strangles (Streptococcus equi) affected mules at Remount Depot,Sargodha (Pakistan)

The prevalence of Streptococcus equi serovar equi (S.equi) in nasal discharge and pus samples from sub‐mandibular lymph nodes in mules at the Remount Depot, Sargodha was examined and total serum proteins, serum albumin, serum globulin and fibrinogen measured. A total of 250 nasal swabs and pus samples were collected from mules and examined microbiologically: 99 (39.6%) were positive for S. equi. A higher occurrence of S. equi was recorded in foals as compared to adults. The concentrations of total serum protein, serum globulin and fibrinogen were significantly increased (P<0.05), while the concentration of serum albumin significantly decreased (P<0.05) in strangles‐affected mules. It was concluded that increased total serum proteins, serum globulin and fibrinogen along with decreased serum albumin were important indicators of infection by S. equi in mules.     

Characterization of a Streptococcus equi ssp. equi Isolate From a Strangles Outbreak in Thailand

Strangles, which is caused by Streptococcus equi ssp. equi, is one of the major infectious respiratory diseases in horses. Knowledge of isolates from different areas of the world is important for investigating the different strains of the disease. In contrast to many other countries, currently little is known about S. equi ssp. equi isolates in Thailand. In 2014, a farm in Thailand imported 20 horses from Europe. Approximately 1 month after arrival, 50% of the horses had developed pyrexia, mucopurulent nasal discharge, and abscesses of the mandibular lymph nodes. Nasal swabs of mucopurulent discharge were sent to a diagnostic laboratory, and two isolates of S. equi ssp. equi were identified. One of the isolates was further characterized using seM gene polymerase chain reaction and sequence analysis. The seM sequence was then compared to the database of PubMLST-seM. It was found to contain SeM allele 48, an allele isolated from horses in the United Kingdom in 2006 and 2010. This result demonstrates the usefulness of SeM allele identification as a tool for investigating the source of related strains and for the epidemiologic study of strangles. To the best of the authors' knowledge, this is the first report of the identification of an SeM allele of the S. equi ssp. equi isolate in Thailand.     

Strangles and its complications

Strangles, caused by the Gram‐positive bacteria Streptococcus equi subspecies equi (S. equi), is a highly contagious upper respiratory infection in horses. The infection is transmitted by inhalation or direct contact with mucopurulent discharge from an infected animal, resulting in fever, depression, and submandibular and retropharyngeal lymph node enlargement that can lead to respiratory distress. Complications include secondary cellulitis at external abscessation sites, guttural pouch empyema and its persistence into the carrier state, purpura haemorrhagica, metastatic abscessation, emergency tracheostomies and rarely secondary S. equi pneumonia or myositis. Control of outbreaks requires strict isolation protocols and hygiene measures. Detection methods of the index case and carrier state are constantly being refined to assist in the identification and prevention of disease perpetuation.     

Novel findings from a beta coronavirus outbreak on an American Miniature Horse breeding farm in upstate New York

This case report describes an outbreak and novel findings associated with a beta coronavirus (BCoV) infection that occurred on an American Miniature Horse (AMH) breeding farm in upstate New York, in January and February of 2013. Twenty-nine AMH and one donkey were present on the farm when the outbreak occurred. One 10-year-old Quarter Horse mare, stabled at a separate location and owned by an employee of the farm, also tested positive. A polymerase chain reaction (PCR) assay for the detection of BCoV was performed at the Animal Health Diagnostic Center (AHDC) at Cornell on all faecal samples. The PCR assay used detects multiple beta coronaviruses, including, but not limited to, equine enteric coronavirus (ECoV). Novel findings regarding this BCoV infection in horses were recognised in this outbreak study. To the authors’ knowledge, this is the largest outbreak of BCoV described thus far in a closed herd on a single premise. The case fatality rate was 0% unlike that described in a previous outbreak of ECoV involving miniature horses and a miniature donkey (Fielding et al. 2015). The morbidity rate was lower in this outbreak than in previously described studies (Oue et al. 2013; Pusterla et al. 2013). This outbreak also demonstrated the potential for BCoV transmission via farm personnel. The duration of shedding of virus in the faeces among some asymptomatic horses in this outbreak was longer than previously described clinical cases of ECoV (Pusterla et al. 2013; Nemoto et al. 2014). This study suggests that asymptomatic animals may play a role in the maintenance of BCoV during an outbreak; therefore, the need for diagnostic testing of both clinically affected and apparently clinically normal horses on a premises followed by appropriate biosecurity and control measures.     

Medical management of Rhodococcus equi infections: A clinical epidemiology perspective

Rhodococcus equi infections cause severe pulmonary disease in foals, affecting animal welfare and increasing production costs in horse-breeding farms. Extra-pulmonary disorders (EPD) are relatively common and can occur independently of pulmonary disease; foals with EPD have a more guarded prognosis. The accompanying paper by Shaw et al. (2021) reports the successful diagnosis and medical treatment of a large abdominal abscess caused by R. equi infection. The authors report on the benefits of using gallium maltolate, a semimetal compound with antimicrobial activity, in combination with traditional R. equi infection antimicrobial treatment (combination of a macrolide and rifampicin). Experimental studies are needed to understand further the benefits of this combined therapy, to evaluate the synergistic effects and if it improves the concentration of antimicrobial drugs into infected tissues. The publication of this case report in Equine Veterinary Education is of clinical importance to equine practitioners when diagnosing and treating R. equi infected foals with or without EPD.     

Clinical observations on an outbreak of strangles

An outbreak of strangles, which occurred during the spring, summer and fall of 1980 on a Standardbred stud farm in eastern Alberta is described. The infective organism, Streptococcus equi, may have been introduced by an outside mare that was brought to the stud for breeding. All of the groups of horses on the farm were affected. For the most part, the disease was allowed to run its natural course. Only severely affected individuals were treated. During the outbreak, the foals were prophylactically treated with penicillin to prevent them from contracting the disease. Ten horses died of complications of strangles infection.     

Hematologic parameters of foals during a strangles epizootic

Hematologic parameters of 23 foals were studied at weeks 0, 2, 4, 6, and 10 following the onset of a strangles epizootic. The epizootic was initiated by group exposure to a foal experimentaUy infected with S, equi. The group consisted of 12 foals previously exposed to an S. equi epizootic 6 months prior, and 11 previously unexposed foals. Seventeen percent of the previously exposed foals and 91% of the unexposed foals developed clinical signs of strangles. Significant increases in mean white blood cell count, neutrophil cell count, fibrinogen concentration, and plasma protein concentration were seen in strangles cases, compared to foals not classified as cases, and were associated with clinical signs. Decreases in packed cell volume, hemoglobin concentration, and red blood cell count, although statistically insignificant, were observed in strangles cases compared to noncases during weeks 4, 6, and 10 and may have biological significance. Similar, but more pronounced, changes were observed in the hematologic parameters of the foal experimentally inoculated with S. equi. The effect of Streptococcus equi infection on the hematology of foals should be considered in their convalescent care.     

Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

Identification of a Streptococcus equi Strain Responsible for Four Outbreaks of Strangles in Colorado

The objective of this study was to use a new subtyping technique to determine the identity of five Streptococcus equi isolates from four outbreaks of strangles in Colorado during 2005−2006. All five of the isolates from the four strangles outbreaks in Colorado contained SeM allele 28. This SeM allele is typical of American isolates of S. equi and has previously been linked with strangles outbreaks in Minnesota in 1994 and Kentucky in 1995. Use of the new S. equi strain subtyping method allowed for characterization of recent S. equi isolates associated with outbreaks of strangles in Colorado. To our knowledge, this is the first report of typing of more recent isolates from North America. This approach, however, has been used in situations in the United Kingdom to differentiate the vaccine strain of S. equi from that of the wild type of the bacteria.     

Streptococcus equi in Equine: Diagnostic and Healthy Performance Impacts

Although the strangles disease of Streptococcus equi was discovered many decades ago in 1,251 by Jordanus Ruffus, it has still remained a major frequently diagnosed infection in horses all over the world. The S. equi subspecies pathogen is known to be often resistant to antibiotic treatment, and it makes the antibiotics inefficient; hence, this review was conducted to study how the disease can be managed. The age-long sign of this infection is the oozing of pus through the mucous and skin membranes. Affected horses lose appetite, develop fever, and become depressed, which result in them losing weight and becoming lethargic, reducing their physical activity especially when they are being raised for athletic and sport purposes. This article reviews various solutions proffered by several researchers about the healthy performance impacts of S. equi in equine.     

Rhodococcus equi pneumonia in an adult horse

A 20‐year‐old, Thoroughbred mare in the fifth month of gestation was examined for weight loss, pyrexia and lethargy. Physical examination, ultrasonography and radiography revealed a severe abscessing pneumonia and a dead fetus. The mare did not respond to symptomatic treatment and died suddenly. Necropsy revealed multifocal pulmonary abscessation. Rhodococcus equi was isolated from the lungs, liver and kidneys. Specific immune function of the mare and presence of the virulence associated protein A (VapA) of the R. equi isolated was not determined. It is likely that immunosuppression is required for systemic R. equi infections in adult horses; however, it is unknown if VapA is necessary to produce disease in adult horses.     

Early pathogenesis of equine Streptococcus equi infection (strangles)

Reasons for performing study: Little is known about entry and subsequent multiplication of Streptococcus equi following exposure of a susceptible horse. This information would have value in design of intranasal vaccines and understanding of shedding and protective immune responses. Objectives: To determine entry points and sites of subsequent replication and dispersion of S. equi at different times after intranasal infection or commingling exposure. Methods: Previously unexposed horses and ponies were subjected to euthanasia 1, 3, 20 or 48 h following intranasal inoculation with biotin labelled or unlabelled S. equi CF32. Some ponies were inoculated with suspensions of equal numbers of CF32 and its mutants lacking capsule, S. equi M‐like protein or streptolysin S. Others were infected by commingling exposure and subjected to euthanasia after onset of fever. Tonsils and lymph nodes were cultured for S. equi and tissues sectioned for histopathological examination and fluorescent microscopy. Results: Tonsillar tissues of both the oro‐ and nasopharynx served as portals of entry. Entry was unexpectedly rapid but involved few bacteria. Small numbers of organisms were detected in tonsillar crypts, in adjacent subepithelial follicular tissue and draining lymph nodes 3 h after inoculation. By 48 h, clumps of S. equi were visible in the lamina propria. At onset of fever, tonsillar tissues and one or more mandibular and retropharyngeal lymph nodes were heavily infiltrated by neutrophils and long chains of extracellular S. equi. Mutant S. equi lacking virulence factors were not seen in draining lymph nodes. Conclusions: Although very small numbers of S. equi entered the lingual and nasopharyngeal tonsils, carriage to regional lymph nodes occurred within hours of inoculation. This observation, together with visual evidence of intracellular and extracellular multiplication of S. equi in tonsillar lymphoid tissue and lymph nodes over the following days, indicates involvement of potent antiphagocytic activity and failure of innate immune defences. Relevance: Future research should logically address the tonsillar immune mechanisms involved including identification of effector cell(s) and antigens.     

Validation of a point-of-care polymerase chain reaction assay for detection of Streptococcus equi subspecies equi in rostral nasal swabs from horses with suspected strangles

This study aimed to validate a point-of-care polymerase chain reaction (PCR) assay for detection of Streptococcus equi subsp. equi (S. equi) in rostral nasal swabs from horses with suspected acute strangles and to compare the results against the molecular gold standard of quantitative polymerase chain reaction (qPCR). Two hundred thirty-two individual swabs of rostral nasal passages were characterized by qPCR as S. equi positive, S. equi subsp. zooepidemicus (S. zooepidemicus) positive, or S. equi and S. zooepidemicus negative. The specificity and sensitivity of the point-of-care PCR assay were 89% and 84%, respectively. The limits of detection of the qPCR assay and the point-of-care PCR analyzer were 3 and 277 eqbE target genes of S. equi, respectively. Overall agreement and short turnaround time make the point-of-care PCR assay a potential molecular diagnostic platform that will enhance the capability of equine veterinarians to timely support a diagnosis of strangles and institute proper biosecurity protocols.     

Guttural pouch empyema caused by Corynebacterium pseudotuberculosis in a pregnant mare

During medical management of mild colic in a 12‐year‐old Quarter Horse, mid‐gestation mare, unilateral purulent nasal discharge from the right nostril was noted. Endoscopic examination revealed guttural pouch empyema. Culture was positive for Corynebacterium pseudotuberculosis and negative for Streptococcus equi ssp. equi. A synergistic haemolysis inhibition titre of 1024 was consistent with C. pseudotuberculosis infection. Treatment included serial lavages and local infusion of antibiotics into the guttural pouches along with a 6‐week course of oral trimethoprim–sulfamethoxazole and rifampicin. Overall, no additional sites of infection were identified and the mare responded well to treatment, delivering a healthy, full‐term foal. This case emphasises that C. pseudotuberculosis, although uncommon, should be considered as a differential for guttural pouch empyema.