Increased Inflammatory Mediators in Horses Naturally Infected with Trypanosoma vivax. A Preliminary Study

The aim of this study was to measure the levels of inflammatory mediators in serum from horses naturally infected with Trypanosoma vivax. Banked serum samples collected during a previously reported T. vivax natural infection were used to analyze proinflammatory cytokines such as interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6), and nitrite/nitrate (NO_x) levels. We evaluated 12 serum samples from horses from a farm in southern Brazil, four of which had parasitological and molecular diagnoses for T. vivax and presented with clinical signs of disease. Cytokines were assessed by quantitative sandwich enzyme-linked immunosorbent assay, and NO_x was measured using the modified Griess method. Levels of IFN-γ, TNF-α, IL-1, IL-6, and NO_x were increased in serum of infected animals compared to that in noninfected animals. Therefore, infection with T. vivax caused an increase in proinflammatory cytokines and nitric oxide content.     

Seroprevalence of Sarcocystis neurona and Its Association With Neurologic Disorders in Argentinean Horses

Equine protozoal myeloencephalitis (EPM) is generally caused by Sarcocystis neurona and can produce substantial economic losses on equine production in America. The aims of the present study were to evaluate the seroprevalence of S. neurona in the main horse-production area of Argentina and associate it with the occurrence of neurologic disorders. Serum samples were collected from 640 horses in nine Argentinean provinces. Most of the samples correspond to animals ≥1.5-year-old from different breeds (n = 628); 12 samples were from younger horses. Further seroprevalence comparison was conducted from the older animals grouped with (n = 148) or without neurologic signs (n = 480). Immunoblot: proteins from 2 × 10⁷S. neurona merozoites were used as antigen on each membrane. Reactivity to antigens with relative mobility of 7, 10, and 16 kDa was considered specific for antibodies against S. neurona; reactivity at 30 kDa was recorded separately. The overall seroprevalence for S. neurona was 26.1% (167/640), and all the provinces had positive horses. Seroprevalence of animals with neurologic signs was greater (P < .001) than what was observed in normal horses (39.2% vs. 22.1%), with an odds ratio of 2.27. Reactivity at 30 kDa was detected in 71% of all samples. This study identified a wide distribution of S. neurona–positive animals in Argentina and horses with neurologic signs having a greater seroprevalence than normal horses. Sarcocystis neurona infection should be considered for early differential diagnosis and treatment of animals with neurologic disorders to decrease the economic impact of EPM in Argentina.     

Hematology and some blood chemical parameters as a function of tick-borne disease (TBD) signs in horses

Two statements provide the context within which this investigation was carried out: (i) Apulia is an enzootic region for tick-borne diseases (TBD); (ii) the principles of enzootic stability and the study of resilient/resistant breeds are among the tools for future pest control strategies. Thus, a trial protocol was established to check the amount of variation in hematologic and biochemical parameters serving as a function of TBD signs in resilient horses. Blood samples were collected from 109 horses belonging to the Apulian native Murgese breed. Hematologic and biochemical parameters were screened and microscopic search for the parasites in the red cells was performed. Differences in clinical manifestations and hematological and biochemical responses were recorded. The following disease classification was used: infection-free horses (class 0), infected horses (presence of Theileria equi and/or Babesia spp in the red cells) without clinical signs (class 1), and horses suffering from clinical disease (class 2). Seventy-one horses (65.1%) showed no TBD signs; hemoparasites were found in 38 horses (34.9%), of which only five (4.6%) showed clinical signs. A significant decreasing trend was shown from class 0 to class 2, where animals with clinical signs presented highly reduced values of some relevant blood parameters. The similarity of the blood values of class 0 and 1 animals is the most striking result of this work and calls for more extensive, detailed research into the mechanisms through which infected horses with no clinical signs of disease can cope with the condition.     

Evidence of host adaptation in Lawsonia intracellularis infections

Background

Lawsonia intracellularis is the causative agent of proliferative enteropathy, an endemic disease in pigs and an emerging concern in horses. Enterocyte hyperplasia is a common lesion in every case but there are differences regarding clinical and pathological presentations among affected species. We hypothesize that host susceptibility to L. intracellularis infection depends on the species of origin of the bacterial isolate. The objective of this study was to evaluate the susceptibilities of pigs and horses to L. intracellularis infection using either a porcine or an equine isolate.

Materials and methods

Twelve foals and eighteen pigs were equally divided into three groups and infected with either a porcine or an equine isolate (10⁹L. Intracellularis/challenged animal), and a saline solution (negative control group). The animals were monitored regarding clinical signs, average of daily weight gain, fecal shedding of the bacteria by PCR and humoral serological response.

Results

Foals infected with the equine isolate developed moderate to severe clinical signs and maintained a lower average of weight gain compared to control foals. Fecal quantitative PCR in equine isolate-infected foals revealed higher amounts of bacterial DNA associated with longer duration of shedding compared with porcine isolate-infected foals. All four foals infected with the equine isolate demonstrated higher IgG titers in the serum compared with porcine isolate-infected foals. In the pig trial, diarrhea and seroconversion were only observed in animals infected with the porcine isolate. Pathological changes typical of proliferative enteropathy were observed in the necropsied foal infected with equine isolate and in the two necropsied pigs infected with the porcine isolate.

Conclusions

Evident clinical signs, longer periods of bacterial shedding and stronger serologic immune responses were observed in animals infected with species-specific isolates. These results show that host susceptibility is driven by the origin of the isolated L. intracellularis strain.     

Seroprevalence of Leishmania sp. Infection in Healthy Horses Housed in Endemic Areas in Tuscany

This study carried out an epidemiological survey of seroprevalence of positive immunofluorescence antibody test (IFAT) results for Leishmania infantum in horses living in Tuscany, where the disease in the dog is endemic and no cases of equine leishmaniasis were observed. Inclusion criteria were (1) horses housed for more than 2 years in endemic areas; (2) horses grazing 24 hours a day outside; and (3) horses living on farms where affected dogs were housed. Two blood samples each were collected from 277 horses from June to October 2011 (T1) and from December 2011 to February 2012 (T2), and IFAT was performed for L. infantum. A dermatologic examination was performed to detect the presence of skin lesions. No animals had skin abnormalities. At T1, 18 of 277 horses had positive results for IFAT, while at the second sampling (T2) 277 of 277 samples were negative. In conclusion, our seroprevalence is lower than that in Spain but higher than that in Greece. Our results suggest the presence of a transient humoral response to L. infantum in horses.     

Equine infectious anemia in mules: virus isolation and pathogenicity studies

There appears to be a lack of information concerning responses of mules to natural infection or experimental inoculation with equine infectious anemia virus (EIAV). In the present study EIAV was isolated from mules, for the first time, and its pathogenicity in naturally infected and experimentally inoculated animals was investigated. Two naturally infected (A and B) and three EIAV free mules (C, D and E) were used for this purpose. Mule A developed clinical signs, whereas mule B remained asymptomatic until the end of the study. Mules C and D were each inoculated with 10ml of blood from mule A and developed signs of the disease; they were euthanatized or died at day 22 and 25 post-inoculation, respectively. Mule E served as a negative control. The virus was isolated from the plasma samples of mules with clinical signs of the disease (A, C and D), but not from the asymptomatic mule B. Both proviral DNA and viral RNA were amplified from blood and tissues of the infected animals by nested polymerase chain reaction (nPCR). Antibodies were not detected in the two experimentally infected mules until their natural death or euthanasia. Clinicopathological and laboratory findings showed that, in mules, EIAV produced clinical signs similar to those observed in horses and ponies. Nested PCR proved to be a rapid, sensitive and specific diagnostic method for the detection of EIAV, regardless of the disease stage.     

A survey of aged horses in Queensland, Australia. Part 2: Clinical signs and owners' perceptions of health and welfare

Objective To describe the prevalence and risk factors for clinical signs of disease and owner-reported health or welfare issues of aged horses in Queensland, Australia. Methods Owners of horses were contacted via an equestrian organisation in Queensland and asked to complete a questionnaire about their horses aged 15 years and older, to gain information about clinical signs observed in the horse and disease history. Owners were asked to identify health or welfare issues they felt were important in aged horses. Results Owners were able to identify many clinical signs of disease, with 83% of horses having at least one reported clinical sign of disease. Muscle stiffness (33%), lameness (28%), ocular discharge (22%), change in hair coat (22%) and skin problems (21%) were the most commonly reported signs. Increasing age was a risk factor for the presence of clinical signs of disease. Ponies had an increased risk of both laminitis and hirsutism, whereas Thoroughbreds had an increased risk of stereotypies, wounds, weight loss and musculoskeletal signs. Owners reported a known disease or disorder in their horses less frequently than the clinical signs of disease. Owners reported weight loss (28%), arthritis/lameness (24%) and teeth/dental care (18%) as the most important health issues, followed by welfare issues (8%). Conclusion Owners demonstrated concern for the health and welfare of their aged horses, but the lack of appropriate recognition of the significance of clinical signs and limited veterinary care suggest a need for increased client communication and veterinarian–client interaction.     

Trypanosoma evansi infection in mainland Spain

An outbreak of Trypanosoma evansi infection that occurred in mainland Spain is described. The outbreak occurred on an equine and camel farm to which dromedary camels from an infected area of the Canary Islands had recently been introduced. One of these camels developed clinical signs and T. evansi was discovered in a blood smear examination. The herd was evaluated in order to determine the extent of the disease. The results showed that 76% of the camels, 35% of the donkeys and 2% of the horses were affected. The animals were isolated and treated using Cymelarsan^® (0.5 mg/kg). After treatment, three blood analysis using parasitological methods revealed negative results. This is the first T. evansi outbreak to have occurred in mainland Spain and the second in mainland Europe, both occurring after the introduction of dromedary camels from the Canary Islands.     

Ross River Virus Infection of Horses: Appraisal of Ecological and Clinical Consequences

Ross River virus (RRV) is a mosquito-borne arbovirus of the genus Alphavirus that causes disease in humans and horses in Australia. A temporal association of RRV infection in horses with clinical signs including pyrexia, malaise, and polyarthralgia has been reported, along with reduced athletic performance, often for extended periods. Despite these reports, disease due to RRV remains somewhat controversial as experimental infection of horses has resulted in obvious viraemia yet minimal signs of clinical disease. The relatively high viraemia demonstrated by horses infected with RRV has led to speculation that they could act as an important reservoir host of the virus, although this remains unclear. This review sought to appraise the existing literature relating to RRV infection of horses and to summarize the ecological and clinical consequences of RRV of relevance to the equine industry and to public health more broadly.     

Outbreak of Lice in Horses: Epidemiology,Diagnosis, and Treatment

The aim of this study is to report an outbreak of lice (Bovicola equi) in horses in southern Brazil, focusing on epidemiology, diagnosis, and treatment. Twenty-three horses in a stud farm presented with intense itching, crusted lesions, focal alopecia, and scaly skin. The outbreak occurred during the winter and probably due to the absence of sanitary barriers on the property. The diagnosis was carried out based on clinical signs associated with macroscopic and microscopic visualization of lice identified as B. equi. The treatment was performed with a single dose of Fipronil (Topline® Merial Brasil, Campinas, São Paulo, Brazil) spray (0.5%), eliminating immature and adult stages of lice. The clinical signs caused by the lice in horses disappeared few days after treatment.     

Death of a horse infected experimentally with Anaplasma phagocytophilum

A 19-year-old horse that was one of a group of six horses infected experimentally with Anaplasma phagocytophilum for a study of the pathogenesis of equine granulocytic ehrlichiosis died suddenly two days after first showing clinical signs of disease. The clinical signs and laboratory findings observed before its death were similar to all those of the other infected horses, and to previous reports of this disease. A postmortem examination revealed widespread haemorrhaging in its internal organs, and vasculitis and thrombosis in the kidneys. These changes are consistent with disseminated intravascular coagulation, which has previously been reported in human beings infected with the presumably identical agent of human granulocytic ehrlichiosis.     

The first nosocomial outbreak of methicillin-resistant Staphylococcus aureus in horses in Sweden

Background

Methicillin-resistant Staphylococcus aureus (MRSA) in animals is a rare finding in Sweden. In horses, MRSA was first detected in a screening survey in 2007. In 2008, six clinical cases occurred in an equine hospital, indicating an outbreak.

Method

All MRSA isolates detected, 11 spa-type t011 and one t064 (n = 12), in infected horses (n = 10) and screening of horses (n = 2) in Sweden from December 2007 to March 2010 were retrospectively analysed with pulsed-field gel electrophoresis (PFGE) using Cfr9I and ApaI restriction enzymes, to study relationship between the isolates. Medical records of infected horses and outbreak investigation notes were scrutinised to monitor the clinical outcome and other aspects of the outbreak.

Results

Eight of the 10 infected horses were linked to one equine hospital and two to another hospital in the same region. The six horses infected with MRSA in 2008 underwent surgery during the period 22 May-7 July in one of the hospitals. Four more infections linked to the two hospitals were notified between 2009 and March 2010.Nine of the 11 spa-type t011 isolates had identical Cfr9I and ApaI PFGE pattern. All six infected horses from 2008 presented with this MRSA. Two t011 isolates differed in one and two bands, respectively, in PFGE.Nine horses suffered from surgical site infections (SSI). No antimicrobials were used following the MRSA diagnosis and the infections cleared. The time from surgery to MRSA diagnosis differed greatly between the horses (range 15-52 days).

Conclusions

Association in time and space of six horses infected with an identical MRSA strain of spa-type t011 confirmed an outbreak. Two isolates found in 2009 and 2010 in the outbreak hospital were closely related to the outbreak strain, indicating one circulating strain. Both spa-type t011 and t064 have been reported in horses in Europe prior to these findings. The observation that the infections cleared although antimicrobials were not used is encouraging for future prudent use of antimicrobials. The time from surgery to bacteriological diagnosis was not acceptable in most cases, as contagious spread was a risk. Sampling when symptoms of infection are noticed and accurate analysis are thus important.     

Occurrence of Trypanosoma evansi in Horses in the State of Minas Gerais,Brazil

Ttrypanosomosis caused by Trypanosoma evansi is a condition that causes significant losses to farmers in endemic areas. This study aimed to report one case of trypanosomiasis in the municipality of Itabira, state of Minas Gerais, Brazil. In November 2010, on a farm with 16 horses, three horses had clinical signs of anemia, limb edema, weight loss, lameness, muscle atrophy, and incoordination of the hind limbs. Trypanosomosis was suspected, and the animals with clinical signs were treated with two doses of diminazene aceturate (7 mg/kg, intramuscular) every 7 days; the treated animals recovered from the clinical symptoms. Blood samples were collected from six horses on the farm, including the three treated animals, to perform polymerase chain reaction specific for T evansi. Four samples were polymerase chain reaction negative, including those collected from three treated horses. However, two other asymptomatic horses were positive for the parasite based on the molecular testing. Based on the results, we concluded that the initial clinical suspect of trypanosomosis was correct, and the treatment used was effective. Probably the disease was introduced to Minas Gerais State through a stallion, which acquired the infection in an endemic area in southern Brazil.     

Immunization with Trichophyton verrucosum Vaccine in Hunter/Jumper and Dressage Horses with Naturally Occurring Trichophyton equinum Infection: A Prospective,Randomized, Double-Blinded,Placebo-Controlled Clinical Trial

The aim of the current study was to investigate the efficacy of Trichophyton verrucosum vaccine in hunter/jumper and dressage horses with trichophytosis caused by Trichophyton equinum (cross immunity). A total of 25 hunter/jumper and dressage horses between the ages of 2 and 14 years, naturally infected with Trichophyton equinum, were enrolled and randomly assigned to either a placebo or an intramuscular lyophilized Trichophyton verrucosum vaccine group. Clinical evaluations were done by the same investigator, who scored clinical healing at the beginning, during, and at the end of the treatment and was blinded to allocation to the groups. At the end of the trial, vaccine treatment significantly decreased the investigator's clinical scores (P < .001), and no significant changes were detected in the placebo treatment group. In all of the horses vaccinated, the lesions improved gradually within 7 to 12 days after vaccination. Complete clinical remission was detected within 28 to 42 days, and all treated horses became culture negative within 25 days of starting treatment. No clinical healing was observed in nine untreated control horses throughout the study. No recurrence was observed in treated horses within 10 months after therapy. Results of the current study indicate that inactivated Trichophyton verrucosum vaccine may be a safe and effective therapy for horses with Trichophyton equinum infection.     

Experimental cross-infections with Ehrlichia phagocytophila and human granulocytic ehrlichia-like agent in cows and horses.

Four cows and four horses were infected experimentally with Ehrlichia phagocytophila, the cause of tickborne fever in ruminants, and with human granulocytic ehrlichia-like agent, a recently discovered species that infects people, horses and dogs in the USA and Europe. They were infected in either order, 30 days apart, to investigate serological cross-reactivity within the Ephagocytophila genogroup. The course of infection was assessed by routine clinical, haematological, serological and polymerase chain reaction (PCR) examinations. Two of the cows infected with Ephagocytophila and two of the horses infected with granulocytic ehrlichia-like agent, developed characteristic signs of ehrlichiosis. When the same animals were infected with their heterologous ehrlichial isolate 30 days later, they did not develop clinical signs of disease. The infection of the other two cows with human granulocytic ehrlichia-like agent and the other two horses with Ephagocytophila, resulted in asymptomatic seroconversion. When the same animals were infected with their homologous ehrlichial isolate 30 days later, they remained asymptomatic and had normal haematological results and negative PCRS until the end of the monitoring period, 60 days after the first infection. In these animals, there was an increase in antibody titre after the second infection which was interpreted as a specific immune response, and as a reactivation of the immune response to the first infection.     

No evidence of horizontal infection in horses kept in close contact with dogs experimentally infected with canine influenza A virus (H3N8)

Background

Since equine influenza A virus (H3N8) was transmitted to dogs in the United States in 2004, the causative virus, which is called canine influenza A virus (CIV), has become widespread in dogs. To date, it has remained unclear whether or not CIV-infected dogs could transmit CIV to horses. To address this, we tested whether or not close contact between horses and dogs experimentally infected with CIV would result in its interspecies transmission.

Methods

Three pairs of animals consisting of a dog inoculated with CIV (10^8.3 egg infectious dose₅₀/dog) and a healthy horse were kept together in individual stalls for 15 consecutive days. During the study, all the dogs and horses were clinically observed. Virus titres in nasal swab extracts and serological responses were also evaluated. In addition, all the animals were subjected to a gross pathological examination after euthanasia.

Results

All three dogs inoculated with CIV exhibited clinical signs including, pyrexia, cough, nasal discharge, virus shedding and seroconversion. Gross pathology revealed lung consolidations in all the dogs, and Streptococcus equi subsp. zooepidemicus was isolated from the lesions. Meanwhile, none of the paired horses showed any clinical signs, virus shedding or seroconversion. Moreover, gross pathology revealed no lesions in the respiratory tracts including the lungs of the horses.

Conclusions

These findings may indicate that a single dog infected with CIV is not sufficient to constitute a source of CIV infection in horses.     

Acute clinical, hematologic, serologic, and polymerase chain reaction findings in horses experimentally infected with a European strain of Anaplasma phagocytophilum

Six horses were experimentally infected by administration of horse blood containing a Swedish strain of Anaplasma phagocytophilum. The polymerase chain reaction (PCR) signal was consistently detected 2-3 days before appearance of clinical signs and persisted 4-9 days beyond abatement of clinical signs, whereas diagnostic inclusion bodies were 1st noted on average 2.6 +/- 1.5 (SD) days after onset of fever. Clinical signs and hematologic changes were largely indistinguishable from those previously reported for diseases caused by A phagocytophilum (formerly Ehrlichia equi--"Californian agent") and the human-derived human granulocytic ehrlichiosis agent. Horses 1st demonstrated antibody response 12-16 days after inoculation, 2 cases of which were still febrile, and serotiters rapidly peaked within 3-7 days of clinical illness. One horse died during the acute stage of disease, but initial clinical signs and hematologic changes were similar to those of other infected horses. This report shows that, despite minor genetic differences, a European equine-derived strain of A. phagocytophilum may be similar in pathogenicity to the Californian agent. The PCR used holds promise to widen the diagnostic window and would also be diagnostic during the initial days of clinical disease when inclusions in neutrophils in blood smears are not yet apparent.     

An acute outbreak of equine dysautonomia (equine grass sickness) in a group of eight Przewalski's horses (Equus ferus [caballus] przewalskii)

Equine grass sickness (EGS) is a neurodegenerative disease affecting grazing equids of which a single case of the chronic clinical presentation has previously been reported in a Przewalski's horse (Equus ferus [caballus] przewalskii). A group of 8 Przewalski's horses were moved to a new enclosure, recently vacated by a group of 4 Eastern kiang (Equus kiang holdereri) that showed no evidence of disease. After 23 days the first Przewalski's horse showed clinical signs of acute EGS including flank sweating, belly kicking, rapid loss of body condition, cessation of faecal passage, nasogastric reflux and mouthing water. It was subjected to euthanasia within 48 h due to lack of therapeutic response. Within 24 h of this first case developing clinical signs, a further 5 Przewalski's horses showed similar clinical signs of acute EGS and were subjected to euthanasia. Post mortem examinations confirmed acute EGS, with all animals demonstrating typical chromatolysis, cytoplasmic hypereosinophilia, cellular swelling, vacuolation, pyknosis and loss of nuclei in approximately 90% of neurones in the cranial cervical and cranial mesenteric ganglia and myenteric and submucosal plexi of the ileum. Two Przewalski's horses within the group showed no clinical signs of disease. No single pathogen was identified as the causal agent, but the epidemiological pattern of the outbreak was typical for that previously reported for acute EGS in domestic equids. All affected animals and the 2 surviving Przewalski's horses had low antibody titres to Clostridium botulinum type C. This is the first report of acute EGS in a herd of Przewalski's horses.     

Subclinical leptospirosis may impair athletic performance in racing horses

The infection by Leptospira in horses, in both its acute disease and subclinical forms, is very common, particularly in endemic regions. Therefore, the objective of this study was to evaluate the effects of subclinical leptospirosis in the athletic performance of racing thoroughbred horses. Athletic performance of 119 racing Thoroughbred horses from Rio de Janeiro, Brazil, was calculated by assigning a point value for the results in racing (performance index (PI)), and serology for leptospirosis was conducted. A total of 85 (71.4?%) horses showed reactive titers (??100), and of which 52 had high titers (34 with 400 and 18 with ??800). Although those animals had high titers against Leptospira, no clinical signs associated with leptospirosis were observed. Seventeen (89.5?%) out of the 19 horses with substandard performance were seroreactive with high titers, in contrast with 35?% of seroreactivity in horses with good athletic performance (P?<?0.0001). Additionally, seroreactivity to leptospirosis was more often observed in horses with substandard athletic performance in contrast to those with good performance (P?<?0.0001, odds ratio 15.8). The Average PI of this group increased to 133?% after treatment (P?<?0.0001). Leptospirosis may impair performance in racing horses, and antibiotic therapy may improve the performance of affected animals.     

A cross-sectional study of geriatric horses in the United Kingdom. Part 2: Health care and disease

Reasons for performing study: Geriatric horses (aged ≥15 years) represent a substantial proportion of the equine population, yet very few studies have investigated the prevalence of diseases within the UK equine geriatric population. Objectives: To describe the provision of routine preventive health care measures, prevalence of clinical signs of disease and the prevalence of owner reported diseases. Additionally, the effect of increasing age on the provision of preventive health care and the presence or absence of clinical signs and disease was assessed. Methods: A cross‐sectional study was conducted, surveying a randomly selected sample of veterinary registered owners with horses aged ≥15 years, using a self‐administered postal questionnaire. Results: As geriatric horses increased in age, there was a reduction in the provision of preventive health care measures, such as vaccination, farrier care and routine veterinary checks. Only 68.7% of horses had received a routine veterinary visit within the previous 12 months. Owners frequently observed clinical signs in their animals, with 77% reporting at least one clinical sign of disease. Increasing age was associated with increased reporting of many clinical signs of disease. Over half (58%) of horses had at least one episode of disease within the previous 12 months, yet only 31% of owners reported that their animal currently suffered from a known disease or disorder. Conclusions and potential relevance: Although owners frequently observed clinical signs in their aged horse, there may be incorrect or under recognition of many diseases and health problems. Reduced frequency of routine preventive health care measures, along with suboptimal owner recognition of health and welfare problems may lead to compromised welfare in the geriatric population.