Persistent development of adomavirus and aquareovirus in a novel cell line from marbled eel with petechial skin haemorrhage.

In Taiwan, a petechial haemorrhage disease associated with mortality has affected marbled eels (Anguilla marmorata). The eels were revealed to be infected with adomavirus (MEAdoV, previously recognized as a polyoma-like virus). In this study, cell line DMEPF-5 was established from the pectoral fin of a diseased eel. DMEPF-5 was passaged >70 times and thoroughly proliferated in L-15 medium containing 2%-15% foetal bovine serum at 20-30°C. Transcripts of neural cell adhesion molecule 1 and nestin genes, and nucleic acids of MEAdoV and a novel reovirus (MERV) in the cells were demonstrated by reverse transcription-polymerase chain reaction analysis. Phylogenetic analysis revealed that the AdoV LO8 proteins mostly relate to adenovirus adenain, whereas MERV is close to American grass carp reovirus in Aquareovirus G, based on a partial VP2 nucleotide sequence. DMEPF-5 cells are susceptible to additional viral infection. Taken together, the marbled eels with the haemorrhagic disease have coinfection with MEAdoV and MERV, and the pathogenic role of MEAdoV and MERV warrants research. DMEPF-5 has gene expression associated with mesenchymal stem and progenitor cells and is the first cell line persistently infected with adomavirus and aquareovirus. DMEPF-5 can facilitate studies of such viruses and haemorrhagic disease.

Reptiles with dermatological lesions: a retrospective study of 301 cases at two university veterinary teaching hospitals (1992-2008).

This retrospective study reviews the medical records of 301 reptiles with dermatological lesions that were examined at the Veterinary Medical Teaching Hospital, University of California at Davis (VMTH-UCD) and the Unité de Dermatologie-Parasitologie-Mycologie, Ecole Nationale Vétérinaire de Nantes (UDPM-ENVN) from 1 January 1992 to 1 July 2008. The most common reptile groups differed between the two hospitals, with lizards being the most common at the VMTH-UCD and chelonians at the UDPM-ENVN. At the VMTH-UCD, boa constrictors (Boa constrictor), ball pythons (Python regius) and other Python species were over-represented, and box turtles (Terrapene carolina) were under-represented in the dermatological lesion caseload. When institutional data were combined, 47% of all reptiles at both institutions with confirmed or suspected cases of sepsis had petechiae, with the highest association seen in chelonians at 82%. Dependent on institution and reptile group, from 29% to 64% of the cases had underlying husbandry issues. Sixty-two per cent of all cases were alive at final status. Veterinarians treating reptiles with skin disease should be aware of the following: (i) that boa constrictors and Python species may be predisposed to dermatological lesions; (ii) that client education is important for proper husbandry; and (iii) that there is a possible association between petechiae and sepsis, especially in chelonians. The conjectural association between certain skin lesions and sepsis remains to be confirmed by systematically derived data that demonstrate a causal relationship between the two.

Ehrlichia canis in dogs in Yucatan, Mexico: seroprevalence, prevalence of infection and associated factors.

The aim of this study was to determine the prevalence of infection, seroprevalence, and factors associated with antibody response to Ehrlichia canis in dogs of Yucatan, Mexico. The study was carried out in four veterinary clinics located in Merida, the capital city of Yucatan, Mexico. Blood samples were obtained from a total of 120 dogs, and each animal was physically examined to determine age and sex, as well as to record any clinical signs of platelet-related bleeding. Blood samples were analyzed for antibodies to E. canis using an ELISA test, and thrombocyte counts were calculated. Blood smears were prepared to detect typical morulae in leukocytes. The prevalence of infection, seroprevalence and associated factors were calculated. A primary screening was performed using 2 x K contingency tables of exposure variables. All variables with P< or =0.20 were analyzed by a logistic-binomial regression. Fifty-three (44.1%) of the 120 dogs were found to be seropositive to E. canis. In six dogs (5.0%) typical morulae of E. canis were observed in monocytes. These six cases were positive for antibodies and were thrombocytopenic. The following factors associated with seropositive animals were: platelet-related bleeding (Yes: OR=10.26, CI=2.50-42.16, P=0.001), thrombocytopenia (Yes: OR=18.91, CI=4.47-80.03, P=0.000) and age (2-4 years: OR=6.77, CI=1.76-25.97, P=0.005; >4 years: OR=4.24, CI=1.04-17.21, P=0.043).

Viral antigen and B and T lymphocytes in lymphoid tissues of gnotobiotic piglets infected with hog cholera virus.

Four 4-day-old gnotobiotic piglets infected intranasally with the Kanagawa/74 strain of hog cholera virus (HCV) did not develop severe illness over a period of 3 weeks. Large amounts of HCV were isolated from the lymphoid tissues and serum at necropsy. After the acute phase, hyperplasia of histiocytes and plasmacytopoiesis were observed in two pigs (killed 14 and 21 days after inoculation). The number of CD4+ and CD8+ T lymphocytes increased significantly and their location was consistent with the site of HCV replication. The results suggest that a CD8+ T-lymphocyte reaction is associated with persistent HCV infection.

Systemic zygomycosis in farmed tilapia fish.

Decreased feed intake and persistent low-level mortality in a production tank of hybrid tilapia (Oreochromis niloticus x Oreochromis mossambicus x Oreochromis aureus) prompted the submission of three affected fish for diagnosis. Consistent macrosopical findings included multifocal dermal haemorrhage, excess abdominal fluid and an enlarged friable liver. On microscopical examination, broad non-septate fungal hyphae and chlamydospores were identified within numerous internal organs, often within and adjacent to blood vessels. The fungal hyphae were readily seen by silver staining (GMS) and the chlamydospores were stained deep magenta by the periodic acid-Schiff reaction. In addition to several species of Gram-negative bacteria, moderate growths of woolly white fungal colonies were obtained from the posterior part of the kidney in two of the three tilapia. These colonies were identified as a Rhizomucor sp. on the basis of the morphological characteristics of the sporulating fungi in culture. This represents the first reported episode of zygomycosis in fish.

Post-transfusion purpura in a pig.

A pig which was being immunised with thrombocytes from another pig developed thrombocytopenia and purpura. This appears to be analogous to the post-transfusion purpura syndrome which has been reported in humans and it is suggested that similar systems in pigs could be useful models for experimental studies on the human condition.

Muscle disorders in the horse: a retrospective study.

Case records of horses with muscle disorders presenting to the Veterinary Medical Teaching Hospital of the University of California, Davis, over a nine year period were evaluated. The objectives of the review were to identify the common myogenic muscle problems and their clinical features. Muscle disease of idiopathic aetiology following exercise was by far the most common condition noted. Other causes of myogenic muscle disorders included congenital, infectious, immune-mediated and nutritional factors.

Unusual patterns of serum antibodies to Streptococcus equi in two horses with purpura hemorrhagica.

An enzyme-linked immunosorbent assay (ELISA) was developed for use in horses to determine serum titers of antibodies of the immunoglobulin classes IgA, IgG, and IgM to Streptococcus equi M-like protein and culture supernatant protein antigens. Serum antibodies were determined in 28 adult horses, including 9 horses with recent S. equi infections, 17 horses without known exposure to S. equi, but without a history of respiratory disease in the preceding 4 months, and 2 horses with clinical purpura hemorrhagica. Serum IgA titers to culture supernatant protein antigen were highest in recently infected horses (P less than 0.001). Serial determinations of antibody titers in the horses with purpura showed that IgG antibodies to both S. equi M-like protein and culture supernatant protein antigens were undetectable initially, but later rose coincidental with clinical recovery from the disease. Possible mechanisms for these findings are discussed.

Haemorrhagic diathesis in neonatal calves: an emerging syndrome in Europe.

In 2008 and 2009 a large number of cases of haemorrhagic diathesis (HD) in neonatal calves were reported in different European countries. In Flanders, 84 cases of neonatal HD in 30 herds were reported in this period. The disease typically affects calves younger than 1 month old from different breed and gender. Prominent clinical signs are cutaneous bleeding, petechiae on all mucosae, melena and often high fever. Early in the disease, the mental state of the animals is uncompromised. The typical haematological finding is pancytopenia, with severe to complete thrombocytopenia being the cause of the increased susceptibility to bleeding. In seven of the affected herds blood samples of calves of the same age group as the clinical case were collected and on six of those farms at least one subclinical case could be identified. Necropsy findings were generalized petechiae, ecchymoses or haemorrhages and variable lymphadenopathy. Histopathology of haemorrhagic lesions revealed multifocal extravasation of red blood cells (haemorrhage) with preservation of tissue architecture and absence of other abnormalities. Total bone marrow aplasia and depletion of all lymphoid tissue was the most prominent finding on histology. Activated macrophages and haemophagocytosis were seen on bone marrow cytology from two live calves. Polymerase chain reaction for bovine viral diarrhoea virus, bluetongue and epizootic haemorrhagic disease virus was negative. Several attempts to isolate a viral agent were unsuccessful.

Molecular analysis of the M protein of Streptococcus equi and cloning and expression of the M protein gene in Escherichia coli.

A Streptococcus equi gene bank was constructed in the bacteriophage lambda gt11 cloning vector, and hybrid phage plaques were screened with S. equi M protein antiserum. A hybrid phage expressing the S. equi M protein (lambda gt11/SEM7) was identified and lysogenized into Escherichia coli Y1089. The cloned M protein appeared in immunoblots as three polypeptides with relative molecular weights of 58,000, 53,000, and 50,000. When reacted with S. equi M protein antiserum in an agar double-diffusion assay, the cloned M protein formed a line of identity with a protein in an acid extract of S. equi. Furthermore, lambda gt11/SEM7 protein inhibited opsonization of S. equi by antiserum to S. equi M protein. In addition, the recombinant protein expressed determinants of the antigen in the immune complexes of purpura hemorrhagica. Native M protein obtained from S. equi and recombinant M protein showed very similar molecular weight distributions on immunoblots, appearing as multiple closely spaced bands with molecular weights ranging from 52,000 to 60,000. Antisera prepared separately against each of the acid-extracted polypeptides shown to be important in serum bactericidal responses (molecular weight, 29,000) and nasopharyngeal local antibody responses (molecular weights, 41,000 and 46,000) of the horse each reacted with all three polypeptides in an acid extract. Moreover, antisera against protoplasts and against recombinant M protein of S. equi also reacted with these polypeptides. These results suggest that the entire M protein molecule of S. equi is present in these preparations and that the fragments in acid extracts carry overlapping segments.

Trypanosoma vivax: disseminated intravascular coagulation in cattle.

Five crossbred cattle infected with Trypanosoma vivax (Likoni) by Glossina morsitans developed capillary haemorrhages at the onset of parasitaemia, followed by the presence of occult blood in faecal samples and eventually melena. Two animals required treatment to survive, on days 13 and 38 respectively. The other three animals cleared their parasitaemias without treatment. Packed cell volume (PCV) levels decreased in all animals to levels ranging from 7.5 to 17%. Relapse in a treated animal initiated marked haemorrhage and a loss of 14 PCV units during a six-day period. Thrombocytopenia was common to all animals, and thrombocytes decreased to levels of 4000/microliters of blood. All animals developed increased levels of fibrinogen and fibrin monomer. Prolonged prothrombin times were found in all animals, and activated partial thromboplastin times were also extended in the two animals with high parasitaemias.