Equine sarcoid of the glans penis with bovine papillomavirus type 1 in a miniature horse (Falabella)

A 23-year-old Falabella gelding kept in Tochigi, Japan, for more than 20 years presented with a recurrent mass of the glans penis that was first noticed about a year earlier. Partial phallectomy was performed with no adjunctive therapy for local regrowth of the mass. The horse was euthanized 3 months after surgery for urinary retention due to suspected regrowth. The resected mass affected the genital and urethral mucosa of the glans penis, and was diagnosed as equine sarcoid by histopathology and identification of bovine papillomavirus (BPV) DNA. Phylogenetic analysis of the BPV genome of the sarcoid showed high sequence homology to BPV type 1 (BPV-1) from Hokkaido, Japan, suggesting a geographical relationship for BPV-1 in Japan.     

Malignant melanoma in a grey horse: case presentation and review of equine melanoma treatment options

A 15 year-old grey Thoroughbred gelding presented for investigation of chronic weight loss and recent onset of respiratory difficulty. Clinical examination confirmed tachypnoea with increased respiratory effort. Thoracic ultrasound examination detected pleural effusion. The dyspnoea was related to the large volume of pleural effusion and, following post-mortem examination, to the presence of a large mediastinal mass. Multiple pigmented masses, likely melanomas, were detected peri-anally. Thoracic radiography, cytological examination of the pleural fluid and a fine needle aspirate of a thoracic mass led to a presumptive diagnosis of malignant melanoma and this was confirmed at post mortem examination. Further metastatic spread to the central nervous system and right guttural pouch was also identified. In conclusion this case manifests the potential malignant behaviour of equine melanomas, and a review of proposed therapies for melanoma treatment highlights the therapeutic options and current areas of research.     

Phylogenetic and structural studies of a novel equine papillomavirus identified from aural plaques

Papillomaviruses (PVs) infect a wide range of animal species and show great genetic diversity. To date, excluding equine sarcoids, only three species of PVs were identified associated with lesions in horses: Equus caballus papillomavirus 1 (EcPV1-cutaneous), EcPV2 (genital) and EcPV3 (aural plaques). In this study, we identified a novel equine PV from aural plaques, which we designated EcPV4. Cutaneous samples from horses with lesions that were microscopically diagnosed as aural plaques were subjected to DNA extraction, amplification and sequencing. Rolling circle amplification and inverse PCR with specific primers confirmed the presence of an approximately 8 kb circular genome. The full-length EcPV4 L1 major capsid protein sequence has 1488 nucleotides (495 amino acids). EcPV4 had a sequence identity of only 53.3%, 60.2% and 51.7% when compared with the published sequences for EcPV1, EcPV2 and EcPV3, respectively. A Bayesian phylogenetic analysis indicated that EcPV4 clusters with EcPV2, but not with EcPV1 and EcPV3. Using the current PV classification system that is based on the nucleotide sequence of L1, we could not define the genus of the newly identified virus. Therefore, a structural analysis of the L1 protein was carried out to aid in this classification because EcPV4 cause lesion similar to the lesion caused by EcPV3. A comparison of the superficial loops demonstrated a distinct amino acid conservation pattern between EcPV4/EcPV2 and EcPV4/EcPV3. These results demonstrate the presence of a new equine PV species and that structural studies could be useful in the classification of PVs.     

Flow cytometric detection of myeloperoxidase in horse neutrophils: a novel technique in equine diagnostic research

Myeloperoxidase (MPO) is a protein of interest due to its involvement in equine pathologies. Until now, results in equine diagnostic research were achieved through extracellular MPO detection. However, studying the cellular MPO content in neutrophils has revealed important insights in human diseases. This study aimed to develop a technique for the specific detection of MPO on the single cell level defining a flow cytometric protocol for the detection of both equine surface-bound and cellular MPO. Both indirect and direct labeling techniques are described which include the comparison of two secondary antibodies and two linking-fluorochromes, respectively.     

Respiratory disease in foals and the epizootiology of equine herpesvirus type 2 infection

The epizootiology of equine herpesvirus type 2 (EHV-2) infection was investigated in Thoroughbred foals on a stud farm which in previous years had suffered economic loss due to respiratory disease. Sixteen pairs of foals and their dams were selected for this study and all of the foals became infected with EHV-2 by two to four months of age. These animals responded serologically to the virus infection as detected by an enzyme-linked immunosorbent assay (ELISA). EHV-2 infection persisted in these foals for two to six months with constant or intermittent virus recovery. This persistent infection stimulated continuous production of antibodies against EHV-2. As soon as the antibody levels reached their peak at five to six months, the isolation rate of EHV-2 from the nasal cavity of these animals decreased, and eventually by nine months of age virus could no longer be recovered. Respiratory disease was observed in ten of the 16 foals; and two severely affected animals died at two months of age. EHV-2 was isolated from both foals at ante and/or post mortem examination. It is postulated that EHV-2, either as an initiating agent or by means of immnunosuppression, caused the respiratory disease observed in these foals.     

Equine penile squamous cell carcinomas are associated with the presence of equine papillomavirus type 2 DNA sequences

Forty cases of equine penile disease were screened with polymerase chain reaction for the presence of papillomaviral DNA. Cases consisted of 20 squamous cell carcinomas (average age of horse, 23.9 years) and 20 non-squamous cell carcinoma diseases (average age of horse, 13.3 years). All horses but one originated from the Northeastern United States. Breeds were not recorded. As based on MY09/MY11 consensus primers, DNA sequences from equine papillomavirus type 2 were amplified from 9 of 20 horses (45%) with penile squamous cell carcinoma and only 1 of 20 horses (5%) with non-squamous cell carcinoma penile disease. Equine papillomavirus type 2 DNA was the only papillomaviral DNA amplified from any of the 40 horses. Tissues from the 10 horses in which papillomaviral DNA was detected by polymerase chain reaction were also screened with in situ hybridization and immunohistochemistry. The presence of papillomavirus was demonstrated in a subset of these by in situ hybridization (6 of 10) and immunohistochemistry (1 of 10). This report describes a possible association between equine penile squamous cell carcinomas and equine papillomavirus type 2. This study is also the first report of equine papillomavirus type 2 infection in North American horses.     

Isolation of equine herpesvirus type 1 from a horse with an acute paralytic disease.

A Standardbred mare became paralyzed shortly after showing signs of an upper respiratory infection. The mare was euthanized and equine herpesvirus type 1 was isolated from the brain and spinal cord.     

Effects of bedding type on compost quality of equine stall waste: implications for small horse farms

Our objective in this study is to compare 4 of the most common bedding materials used by equine operations on the chemical and physical characteristics of composted equine stall waste. Twelve Standardbred horses were adapted to the barn and surrounding environment for 2 wk before the start of the study. Groups of 3 horses were bedded on 1 of 4 different bedding types (wood shavings, pelletized wood materials, long straw, and pelletized straw) for 16 h per day for 18 d. Stalls were cleaned by trained staff daily, and all contents removed were weighed and stored separately by bedding material on a level covered concrete pad for the duration of the study. Compost piles were constructed using 3 replicate piles of each bedding type in a randomized complete block design. Each pile was equipped with a temperature sensor and data logger. Water was added and piles were turned weekly throughout the 100-d compost process. Initial and final samples were taken, dried, and analyzed for DM mass, OM, inorganic nitrogen (nitrate-N and ammonium-N), electrical conductivity, and soluble (plant-available) nutrients. Data were analyzed using the GLM procedure, and means were separated using Fischer's protected LSD test (P < 0.05). No significant temperature differences were observed among the bedding materials. The composting process resulted in significant reductions (P < 0.05) in DM mass for each of the 4 bedding materials. The composting process resulted in significant reductions (P < 0.05) in OM and C:N ratio for all 4 bedding materials. The composted long straw material had greater concentrations of total Kjeldahl nitrogen (P < 0.05), nitrate-N (P < 0.05), and ammonium-N (P < 0.05) than the composted wood shavings. This study demonstrated that incorporating a simple aerobic composting system may greatly reduce the overall volume of manure and yield a material that is beneficial for land application in pasture-based systems. The straw-based materials may be better suited for composting and subsequent land application; however, factors such as suitability of the bedding material for equine use, material cost, labor, and availability must be considered when selecting a bedding material.     

A propagating epizootic of equine infectious anemia on a horse farm

An epizootic of equine infectious anemia (EIA) involved 35 horses on a farm in south Georgia. During a 126-day period, 21 of these horses became seropositive for EIA. After the initial diagnosis in July, the horses were tested every 7 to 10 days. At least one additional horse was found to be seropositive on each testing day. As soon as they were determined to be seropositive, the horses were removed from the herd and sent to slaughter. The removal of the seropositive horses, however, did not stop the epizootic. We believe the initial infection was from a 7-year-old stallion that recently had been purchased or from 1 of 2 mares that were seropositive for EIA on the first test. None of the horses had been tested for EIA at the time of purchase or within 60 days before the epizootic.     

Effects of ketanserin on hypergranulation tissue formation, infection, and healing of equine lower limb wounds

The objective of this multicentre, randomized, controlled field study was to determine the efficacy of ketanserin gel in preventing exuberant granulation tissue formation (hypergranulation) and infection in equine lower limb wounds. Horses and ponies (n = 481) with naturally occurring wounds were randomized to either topical treatment with ketanserin gel (n = 242) or a positive control (Belgium, Germany: ethacridin lactate solution, n = 120; France, United Kingdom: malic, benzoic, and salicylic acid [MBS] cream, n = 119). Treatment continued until the wound healed (success), formed hypergranulation tissue (failure), or became infected (failure). Treatment was terminated after 6 months in all remaining animals. Ketanserin was successful in 88% of cases. Wounds treated with ketanserin were 2 and 5 times more likely to heal successfully than were those treated with MBS or ethacridin lactate, respectively. Ketanserin gel is thus more effective than these standard treatments in preventing hypergranulation tissue and infection of equine lower limb wounds.     

Multiple glomus tumors of the urinary bladder in a cow associated with bovine papillomavirus type 2 (BPV-2) infection

We report a case of multiple glomus tumors associated with bovine papillomavirus type 2 (BPV-2) infection in the urinary bladder of a 13-year-old cow suffering from severe chronic enzootic hematuria. Macroscopically, multiple submucosal reddish nodules were seen swelling the vesical mucosa. Histologically, neoplastic proliferation was characterized by the presence of numerous blood vessels. These were lined by normal endothelial cells surrounded by round epithelioid cells with central nuclei, prominent nucleoli, acidophilic cytoplasm, and well-defined cytoplasmic borders. Tumor cells were distributed around open vascular lumina and in perivascular spaces. They were immunohistochemically positive for actin and vimentin and negative for cytokeratins, desmin, and factor VIII-related antigen. On the basis of these findings, this tumor was diagnosed as glomus tumor, a neoplasm not previously reported in cattle and exceedingly rare in animals. BPV-2 DNA was amplified from the formalin-fixed, paraffin-processed tissue specimens obtained by laser capture microdissection. This report widens the spectrum of mesenchymal tumors of the bovine urinary bladder. Finally, the microscopic pattern of tumor described here shares striking morphologic and immunohistochemical similarities with the angiomatous form of glomus tumor known to occur in man.     

Comparison of methods for the diagnosis of equine herpesvirus type 1 infection

The objective of the investigations was to study the occurrence of the equine herpesvirus type 1 (EHV-1) infection in aborted equine fetuses and in newborn foals and to compare the sensitivity of virus isolation, immunohistochemistry and histology in 101 cases and of fetal serology in 68 cases in the diagnosis of the infection. Out of the 93 aborted equine fetuses and 8 weak foals, 15 (14.9%) (14 fetuses and 1 foal) proved to be EHV-1 infected by immunohistochemical and 13 (12.9%) by virological investigation. Characteristic microscopic changes were seen in several organs in all cases, while intranuclear inclusion bodies could be found only in 25 (35.2%) of the 71 virus-positive tissue samples. Four (5.9%) cases proved to be positive by fetal serological investigation, but none of these cases showed any EHV-1 specific lesions and in none of these cases could the virus be detected by virus isolation or by immunohistochemistry. According to the results, fetal serology does not seem to be a useful test in virus-positive cases, while the immunohistochemical method seems to be a reliable and a slightly more sensitive method than virus isolation in the diagnosis of EHV-1 infection.     

Bovine papillomavirus type-2 (BPV-2) infection and expression of uroplakin IIIb, a novel urothelial cell marker, in urinary bladder tumors of cows

Several immunohistochemical markers have been used to define the differentiation pattern of urothelial cell tumors of the urinary bladder. We investigated the expression of the recently characterized uroplakin (UP) IIIb, an urothelium-specific and differentiation-dependent protein, in 39 urothelial tumors of the urinary bladder in cows that had suffered from chronic enzootic hematuria for several years. Bovine papillomavirus type 2 DNA was amplified and UP IIIb protein was detected in all these tumors. In papillomas and papillary carcinomas, UP IIIb expression was mostly seen as superficial staining; luminal and peripheral patterns were also observed. In nonpapillary carcinomas, UP IIIb appeared to define clearly the cell membrane lining intercellular and intracellular lumina as well as the cell borders in deeper cell layers. In benign and malignant lesions, an intracytoplasmic immunoreactivity was also detected. Coarse intracytoplasmic UP IIIb-positive material close to the nucleus occurred in some malignant cells. Focally strong membraneous immunostaining that marked single cells with complete ringlike peripheral pattern was seen. Although UP IIIb expression does not seem to correlate with the biological behavior of urothelial tumors, it appears to be a highly sensitive marker for bovine urothelial tumors.     

The development of equine immunity: Current knowledge on immunology in the young horse

The development of equine immunity from the fetus to adulthood is complex. The foal's immune response and the immune mechanisms that they are equipped with, along with changes over the first months of life until the immune system becomes adult‐like, are only partially understood. While several innate immune responses seem to be fully functional from birth, the onset of adaptive immune response is delayed. For some adaptive immune parameters, such as immunoglobin (Ig)G1, IgG3, IgG5 and IgA antibodies, the immune response starts before or at birth and matures within 3 months of life. Other antibody responses, such as IgG4, IgG7 and IgE production, slowly develop within the first year of life until they reach adult levels. Similar differences have been observed for adaptive T cell responses. Interferon‐gamma (IFN‐γ) production by T helper 1 (Th1)‐cells and cytotoxic T cells starts shortly after birth with low level production that gradually increases during the first year of life. In contrast, interleukin‐4 (IL‐4) produced by Th2‐cells is almost undetectable in the first 3 months of life. These findings offer some explanation for the increased susceptibility of foals to certain pathogens such as Rhodococcus equi. The delay in Th‐cell development and in particular Th2 immunity during the first months of life also provides an explanation for the reduced responsiveness of young horses to most traditional vaccines. In summary, all immune components of adult horses seem to exist in foals but the orchestrating and regulation of the immune response in immature horses is strikingly different. Young foals are fully competent and can perform certain immune responses but many mechanisms have yet to mature. Additional work is needed to improve our understanding of immunity and immune regulation in young horses, to identify the preferred immune pathways that they are using and ultimately provide new preventive strategies to protect against infectious disease.