High prevalence of bovine papillomaviral DNA in the normal skin of equine sarcoid-affected and healthy horses

Bovine papillomavirus (BPV), the causative agent of papillomas in cattle, has been shown to play a major role in the pathogenesis of equine sarcoids in horses. BPV has also been detected occasionally in normal equine skin. In this study, presence and activity of BPV in normal skin and peripheral blood of 4 groups of horses were evaluated: sarcoid-affected horses, horses living in contact with sarcoid-affected horses, horses living in contact with papilloma-affected cattle and control horses. From each horse, 3 samples on 4 locations were collected: a swab of the intact skin surface and both a swab and a biopsy after decontamination. BPV DNA was found in the normal skin of 24 of 42 horses (57%). Mainly sarcoid-affected horses and horses living in contact with cattle were carriers (73%), but BPV DNA was also detected in 50% of the horses living in contact with sarcoid-affected horses and in 30% of the control population. BPV mRNA was detected in 58% of the samples positive for BPV DNA, although in a much lower quantity compared to sarcoids. In most of the BPV DNA positive samples mild acanthosis, slight basophilic cytoplasmic swelling of the epidermal layers and/or thickening of the basal membrane were noticed, but these observations were also present in several BPV DNA negative normal skin samples. BPV DNA could not be detected in peripheral blood. These findings suggest latent infection and a wide-spread occurrence of BPV in the horse population.     

P-80 Clinical aspects of chronic progressive pastern dermatitis in Belgian draft horses

The aim of this study was to evaluate the fungi present on the skin surface in healthy horses with a particular interest in Malassezia on predisposed skin areas. Twenty-five mares were included, each sampled on nine skin areas including those most likely contaminated by Malassezia : nostrils, lips, ear pinnae, base of the mane, pasterns, pectoral areas, lateral thorax, udder and perianal areas. Two sterile carpets were applied to the skin, and then used for inoculation of agar (Sabouraud's plus cycloheximide and chloramphenicol), both with and without lipids for isolation of different species of Malassezia . This method is currently successful in isolating Malassezia in dogs. Cultures were incubated at 32°C, observed on day 5 for Malassezia , and then maintained at 27°C for complementary morphological identifications. From 223 cultures on standard medium, only seven remained sterile. The most common fungi were filamentous (number of positive samples): Scopulariopsis (156), Penicillium (79), Acremonium (67), Aspergillus (53), Paecilomyces spp. (38), and Stachybotrys (33). Interestingly, Microsporum gypseum was also isolated (14 samples from seven areas) from 10 horses. Yeast or yeast-like fungi were Geotrichum (52), Trichosporon (21), and an additional 33 samples being positive for yeast other than Malassezia . Results from the other 223 cultures on lipid-rich agar were similar. Most of the fungi were isolated from the nine areas selected, but Malassezia was never isolated from 446 cultures.
Funding: Self-funded.     

Transient hypogammaglobulinaemia of the young in a Thoroughbred yearling

A 12-month-old Thoroughbred colt was presented for chronic recurrent respiratory disease characterised by persistent nasal discharge, intermittent pyrexia and coughing of several months' duration. These episodes were responsive to empirical antimicrobial therapy but returned once therapy was discontinued. History, physical examination and initial diagnostics were suggestive of bacterial pneumonia and possible underlying immunodeficiency. Serum immunoglobulin analysis and peripheral blood lymphocyte phenotyping revealed low serum IgG and IgM concentrations. In this 12-month-old colt, low serum IgG concentration was the most diagnostic parameter and supported a presumptive diagnosis of transient hypogammaglobulinaemia of the young. The colt responded favourably to repeated treatment with antimicrobials for bacterial pneumonia, and serial immunologic testing over the following months showed gradual increase in serum IgG and IgM concentrations, supporting the diagnosis of delayed development of humoral immunity in this patient.     

Effect of ovotransferrin and lactoferrins on Chlamydophila psittaci adhesion and invasion in HD11 chicken macrophages

The effect of ovotransferrin (ovoTF), human lactoferrin (hLF) and bovine lactoferrin (bLF) on the obligate intracellular pathogen Chlamydophila (Cp.) psittaci was evaluated using a model of Buffalo Green Monkey kidney (BGM) cells and HD11 chicken macrophages as artificial hosts. Firstly, the effect of transferrins on the infectivity of the bacteria was evaluated. Pre-incubation of Cp. psittaci with 0.5 to 5 mg/mL ovoTF prior to infecting BGM cells significantly lowered the infection rate (P < 0.05). For both lactoferrins, the infection rate could only be reduced with 5 mg/mL, albeit not significantly as compared to the infection rate created by the untreated bacteria. Secondly, transferrins were tested for their ability to influence bacterial adhesion and entry in HD11 cells. Maximal non-cytotoxic and non-bactericidal concentrations of 0.05 mg/mL ovoTF and 0.5 mg/mL hLF and bLF were used. Overall, ovoTF was more effective than human and bovine LF in inhibiting bacterial irreversible attachment and cell entry and the latter was accompanied by a dose-dependent reduction of actin recruitment at the bacterial entry site. However, once bacteria had entered HD11 cells, transferrins had apparently no effect on intracellular replication. The present findings suggest a possible role for transferrins and especially ovoTF, in preventing avian Cp. psittaci infections.     

Antibodies to elastin peptides in sera of Warmblood horses at different ages

REASONS FOR PERFORMING STUDY: Early diagnosis and monitoring progression of chronic diseases in elastin-rich tissues, such as chronic progressive lymphoedema in draught horses and chronic obstructive pulmonary disease (COPD) is still a real challenge in the horse. Use of an enzyme-linked immunosorbent assay (ELISA) to detect anti-elastin antibody (AEAb) levels might be useful to assess the status of such diseases. Baseline levels, representing physiological breakdown of elastin in normal horses, are not available at present. HYPOTHESIS: Levels of AEAb in healthy horses are generally low and follow the same age-related pattern as found in man. Therefore, elevation of AEAb levels in serum can be used to evaluate pathological elastin breakdown in elastin-rich tissues. METHODS: Sera of 84 clinically healthy Warmblood horses were evaluated for the presence of AEAbs by means of a modified version of an ELISA technique used in man. The horses were divided in 5 age groups: A) < 4 months; B) 4-23 months; C) 2-3 years; D) 4-10 years; and E) > 11 years. RESULTS: Antibodies to elastin were found in all equine serum samples tested. Their levels were lowest in Group A, low in Groups B and E and highest in animals age 2-10 years. CONCLUSIONS: Measuring AEAbs in serum of horses by an ELISA technique proved to be possible and levels were stable during well-defined life stages. POTENTIAL RELEVANCE: Changes in AEAb levels are expected to be useful for early diagnosis and for monitoring progression of diseases that affect elastin-rich tissues, such as chronic progressive lymphoedema and COPD.     

Antibodies to elastin peptides in sera of Belgian Draught horses with chronic progressive lymphoedema

REASONS FOR PERFORMING STUDY: Chronic progressive lymphoedema (CPL) is a recently recognised disease of the lymphatic system characterised by lesions in the skin of the lower legs in several draught horse breeds, including the Belgian Draught hourse. Clinical signs slowly progress and result in severe disfigurement of the limbs. Ideally, supportive treatment should be started early in the disease process. However early diagnosis and monitoring progression of CPL is still a challenge. HYPOTHESIS: Elastin changes, characterised by morphological alterations as well as increased desmosine levels, in the skin of the distal limbs of horses affected with CPL are probably associated with a marked release of elastin degradation products, which elicit production of circulating anti-elastin antibodies (AEAbs) in the serum. An enzyme-linked immunosorbent assay (ELISA) for detection of serum AEAbs may document elastin breakdown. METHODS: An ELISA technique was used to evaluate levels of AEAbs in sera of 97 affected Belgian Draught horses that were clinically healthy except for possible skin lesions, associated with CPL in their distal limbs. The horses were divided into 5 groups according to the severity of these skin lesions: normal horses (Group 1, n = 36), horses with mild lesions (Group 2, n = 43), horses with moderate lesions (Group 3, n = 8), horses with severe lesions (Group 4, n = 10) and, as a control, healthy Warmblood horses, unaffected by the disease (Group 5, n = 83). RESULTS: Horses with clinical signs of CPL had significantly higher AEAb levels compared to clinically normal Belgian Draught horses and to healthy Warmblood horses. These levels correlated with severity of lesions. CONCLUSIONS: CPL in draught horses is associated with an increase of serum AEAbs. POTENTIAL RELEVANCE: Evaluation of serum levels of AEAbs by ELISA might be a useful diagnostic aid for CPL. Pathological degradation of elastic fibres, resulting in deficient support of the distal lymphatics, is proposed as a contributing factor for CPL in Belgian Draught horses.     

Evaluation of FOXC2 as a candidate gene for chronic progressive lymphedema in draft horses

Chronic progressive lymphedema (CPL) is a debilitating condition identified in Clydesdales, Shires and Belgian draft horses and results in progressive swelling of the lower legs associated with the development of thick skin folds, ulcerations, fibrosis and marked hyperkeratosis. The result is severe discomfort and recurrent secondary infection, often requiring euthanasia. Due to the delayed onset, many horses are bred prior to diagnosis. CPL has only been documented in three related draft horse breeds, suggesting a genetic cause. Determining the molecular basis would enable owners to test horses prior to breeding and facilitate the elimination of CPL. Mutations in the FOXC2 gene cause a comparable condition in humans, lymphedema-distichiasis. This gene was sequenced in affected and unaffected draft horses and a control horse. Four single nucleotide polymorphisms (SNPs) were identified in unaffected draft horses and the control horse, indicating that they were not associated with CPL. A fifth SNP was seen in a single affected draft horse and the control horse. Since it was not seen in all affected draft horses, this SNP is not associated with the CPL phenotype.     

Analysis of Andean blackberry (Rubus glaucus) production models obtained by means of artificial neural networks exploiting information collected by small-scale growers in Colombia and publicly available meteorological data

The Andean blackberry (Rubus glaucus) is an important source of income in hillside regions of Colombia. However, growers have little reliable information on the factors that affect the development and yield of the crop, and therefore there is a dearth of information on how to effectively manage the crop. Site specific information recorded by small-scale producers of the Andean blackberry on their production systems and soils coupled with publicly available meteorological data was used to develop models of such production systems. Multilayer perceptrons and Self-Organizing Maps were used as computational models in the identification and visualization of the most important variables for modeling the production of Andean blackberry. Artificial neural networks were trained with information from 20 sites in Colombia where the Andean blackberry is cultivated. Multilayer perceptrons predicted with a reasonable degree of accuracy the production response of the crop. The soil depth, the average temperature, external drainage, and the accumulated precipitation of the first month before harvest were critical determinants of productivity. A proxy variable of location was used to describe overall differences in management between farmers groups. The use of this proxy indicated that, even under essentially similar environmental conditions, large differences in production could be assigned to management effects. The information obtained can be used to determine sites that are suitable for Andean blackberry production, and to transfer of management practices from sites of high productivity to sites with similar environmental conditions which currently have lower levels of productivity.     

Multiple mandibular ossifying fibromas in a yearling Belgian Draught horse filly

A yearling Belgian Draught horse filly was presented with a history of bilateral, progressive, painful swelling of the mandible, fever and anorexia. Radiographs and computed tomographic images showed an expansile lesion with a coalescing, well‐defined cystic pattern at the level of the horizontal ramii of the mandible. The histopathological findings confirmed the diagnoses of bilateral, multiple ossifying fibroma. This report indicates that ossifying fibroma should be included in a differential diagnosis if multiple expansile lesions of the jaw are present.     

Molecular characterization and pathogenicity of Pythium species associated with damping-off in greenhouse cucumber (Cucumis sativus) in Oman

A study was undertaken in 2004 and 2005 to characterize pathogens associated with damping-off of greenhouse-grown cucumber seedlings in 13 districts in Oman. Identification of Pythium to the species level was based on sequences of the internal transcribed spacer (ITS) of the ribosomal DNA. Of the 98 Pythium isolates collected during the survey, Pythium aphanidermatum , P. spinosum , P. splendens and P. oligandrum accounted for 76%, 22%, 1% and 1%, respectively. Pythium aphanidermatum was isolated from all of the districts, while P. spinosum was isolated from seven districts. Pathogenicity tests showed inter- and intraspecific variation in aggressiveness between Pythium species. Pythium aphanidermatum , P. spinosum and P. splendens were found to be highly aggressive at 25°C. However, the aggressiveness of P. spinosum decreased when the temperature was raised to 30°C, which was found to correspond to the lower frequency of isolation of P. spinosum in the warmer seasons, compared to the cooler time of the year. Pythium aphanidermatum exhibited limited intraspecific variation in the sequences of the ITS region of the rDNA and showed 100% similarity to the corresponding P. aphanidermatum sequences from GenBank. The ITS sequence data, as well as morphological characteristics of P. spinosum isolates, showed a high level of similarity within and between P. spinosum and P. kunmingense , and suggested that the two species were synonymous. This study represents the first report of P. spinosum, P. splendens and P. oligandrum in Oman.