Application of cloned fragments of equine herpesvirus type-1 DNA for detection of virus-specific DNA in equine tissues

Tissue specimens obtained from equine herpesvirus-1 (EHV-1), subtype 1-infected aborted foetuses were analysed for the presence of virus DNA by means of Southern blot and dot blot hybridisations. The specificity of the methods was confirmed although the sensitivity was inferior to classical techniques such as virus isolation. However, the possibility of detecting the state of the virus DNA and the ability to distinguish between subtypes were important features, and the dot blot method was shown to have potential for a rapid diagnostic test. This report demonstrates some potential practical applications of hybridisation methods for studying the pathogenesis and epidemiology of EHV-1 but also reveals limitations of the techniques.     

First case report of Sarcocystis neurona-induced equine protozoal myeloencephalitis in Japan

Equine protozoal myeloencephalitis developed in a three-year-old male Thoroughbred racehorse imported from the United States. The animal showed astasia five days after the onset of ataxia. Histopathologically, focal nonpurulent myelitis accompanied by hemorrhage and perivascular infiltration was observed in the fourth and fifth cervical spinal cord. Immunohistochemically, shizonts were occasionally observed and were positive for anti-Sarcocystis neurona (S. neurona) antiserum. S. neurona-specific antibodies were detected in the serum and cerebrospinal fluid by Western blot. This is the first equine protozoal myeloencephalitis case in Japan.     

First case report of ultrastructural cutaneous abnormalities in equine atopic dermatitis

Atopic dermatitis (AD) is a common skin disease that affects humans and animals. Skin impairment has been described in human and canine AD. Equine AD is recognized in practice but little is known about its pathogenesis. As remarkable similarities exist across species in terms of cutaneous manifestations of AD, it was speculated that skin abnormalities may also exist in atopic horses. This case report describes the ultrastructure of the stratum corneum of two normal and two atopic horses. Biopsies were taken from sites predisposed to AD and examined using electron microscopy. Stratum corneum in normal samples was compacted with organized lipid lamellae while in atopic samples disorganized lipid lamellae, retained lamellar bodies and amorphous lipids were found. These changes are very similar to what reported in AD in other species. It is currently unknown whether these abnormalities in atopic horses are primary or secondary and their importance in allergen penetration.     

Physicochemical properties of akabane virus: A member of the simbu arbovirus group of the family bunyaviridae

The multiplication of Akabane virus was not inhibited in the presence of 5-iodo-2′-deoxyuridine, indicating the presence of RNA. The virus was considered to have an envelope, as it was sensitive to ether and chloroform. It was readily inactivated by deoxycholate and trypsin, but was not precipitated by protamine sulphate. The virus was very labile at pH 3 and also rather heat-labile. Akabane virus was readily filtered through membrane filters of 200 or 100-nm pore size, but not through 50-nm filters. Equilibrium centrifugation in a CsCl density gradient gave a peak of infectivity and hemagglutinin at a density of 1.22 g/ml. The peak fractions thus obtained contained numerous virus particles, roughly spherical, variable in size, 70 to 130 nm in diameter, and mostly having a ragged, closely adherent envelope with projections, when examined, following phosphotungstic acid negative staining, in an electron microscope.     

First genetic characterization of equine adenovirus type 1 (EAdV-1) in Turkey

Equine adenovirus type 1 (EAdV-1) is a cause of repiratory tract infection in equids. In present study for the first time in Turkey, the prevalence of EAdV-1 in nasal swab samples obtained from horses showing respiratory symptoms was investigated by polymerase chain reaction (PCR), and molecular characterization of the hexon gene detected in the Turkish (TR) strain was performed. Overall, the prevalence of EAdV-1 was found low (1.4%) as indicated by a positive PCR reaction from the nasal swab extracts tested. Phylogenetic analysis based on the partial sequences of the hexon gene of a TR-EAdV-1 strain with those of previously isolated AdVs from different mammals and an EAdV-1 M1 strain showed that the EAdV-1 strains were placed into a unique cluster. Although the TR-EAdV-1 strain was closely related to CAV-1, CAV-2 and bat adenovirus reference strains, larger-scale studies are necessary to better understand the molecular epidemiology and population structure of EAdV-1 in Turkey.     

Topology and biological function of enterovirus non-structural protein 2B as a member of the viroporin family

Viroporins are a group of transmembrane proteins with low molecular weight that are encoded by many animal viruses. Generally, viroporins are composed of 50–120 amino acid residues and possess a minimum of one hydrophobic region that interacts with the lipid bilayer and leads to dispersion. Viroporins are involved in destroying the morphology of host cells and disturbing their biological functions to complete the life cycle of the virus. The 2B proteins encoded by enteroviruses, which belong to the family Picornaviridae, can form transmembrane pores by oligomerization, increase the permeability of plasma membranes, disturb the homeostasis of calcium in cells, induce apoptosis, and cause autophagy; these abilities are shared among viroporins. The present paper introduces the structure and biological characteristics of various 2B proteins encoded by enteroviruses of the family Picornaviridae and may provide a novel idea for developing antiviral drugs.     

First recovery of A/equine/Fontainebleau/1/79 influenza viruses in Italy

In Italy epizootics of equine influenza often occur, but no virus isolation has been reported since 1971. This paper describes the antigenic and biochemical characterization of two equine influenza viruses isolated in Italy from 1985 to 1989. The virus isolates were shown to differ antigenically from earlier strains of the same subtype, A/equine/Miami/1/63 (H3N8). Monoclonal antibody analysis showed that the haemagglutinins of these strains were serologically indistinguishable from A/equine/Fontainebleau/1/79, a variant of A/equine/Miami, never isolated in Italy before. One of the two virus isolates was obtained from a horse immunized with a bivalent inactivated influenza vaccine, not containing A/equine/Fontainebleau/79 antigens.

The vaccine failure underlines the importance of antigenic relatedness between currently circulating viruses and vaccine strains. Therefore, to improve the protection afforded by equine immunization, the vaccine composition should be decided according to the results of a virological surveillance activity, systematically conducted among horses.     

Clinical, viral, and genetic evaluation of equine degenerative myeloencephalopathy in a family of Appaloosas

A clinical, viral, hematologic , and genetic study was conducted over a 4-year period on a family of Appaloosas with high incidence of clinical ataxia and pathologic features of equine degenerative myeloencephalopathy. Marginal to deficient serum vitamin E (alpha-tocopherol) and blood selenium values were the only other consistent antemortem abnormalities in the affected horses. Members of this family were all descendants of a clinically normal mare and were raised in 3 separate environments with variable quality of feed. All horses had access to pasture grasses. Normal chromosomal karyotypes were found in 11 affected and/or related horses examined. Equine herpesvirus type 2 was isolated from 4 of the horses, but evidence for a role of this virus in the pathogenesis of the disease was not found. The role of antioxidant deficiency in the pathogenesis of neurologic dysfunction in this equine family and in others reported to be affected with equine degenerative myeloencephalopathy remains speculative.     

Histomorphology of the equine cervix

The cervix provides an effective, morphological barrier between the uterus and the outside world. The aim of this study was to characterize the epithelial morphology and the vascular structures of the equine cervix along the longitudinal and horizontal axis in more detail by light microscopy. For this purpose, cervical tissue specimens that had been removed from five different regions along the caudocranial axis of 10 genitally healthy mares were available. The histological staining was carried out with haematoxylin-eosin, azan according to Heidenhain, periodic acid-Schiff reaction and resorcinfuchsin. An average epithelial cell height of 17.5 +/- 1.7 microm is measured, there being differences in the various areas of the mucosal folds and along the longitudinal axis of the cervix. Three types of cells can be differentiated morphologically. Contrary to the data in the literature, in a large number of cells the free cell membrane shows a clearly discernible border of kinocilia along the total cervical canal. The deep layers of the lamina propria mucosae show pronounced vascularization (46.3 +/- 25.1 vessels/mm2) that mainly consists of veins and venules. Support of the occlusive function of the cervical canal in the form of a cavernous body is assumed to be the function of this vascular plexus.     

Isolation of Fukuoka virus, a member of the Kern Canyon serogroup viruses of the family Rhabdoviridae, from cattle.

Four virus strains with identical antigenic properties were isolated from blood samples of 4 sentinel calves having a fever and leukopenea in cultures of HmLu-1 cells derived from baby hamster lung. The virus was identified as Fukuoka virus, classified as a member of the Kern Canyon serogroup viruses of the family Rhabdoviridae, on the basis of its antigenic properties.