RESUMEN
Gaucher disease, an autosomal recessive lysosomal storage disorder caused by mutations in the ß-glucocerebrosidase gene, was recently discovered in sheep on a "Southdown" sheep stud in Victoria, Australia. Clinical signs include neuropathy, thickened leathery skin, and ichthyosis, with lambs unable to stand from birth. Affected lambs were found to be deficient in glucocerebrosidase activity, and mutational analysis found them to be homozygous for the missense mutations c.1142G>A (p.C381Y) and c.1400C>T (p.P467L). In addition, four silent mutations were detected (c.777C>A [p.Y259Y], c1203A>G [p.Q401Q], c.1335T>C [p.I445I], c.1464C>G [p.L488L]). The human equivalent [C342Y] to the C381Y mutation leads to an acute neuronopathic phenotype in patients. Identification of an acute neuronopathic form of Gaucher disease in sheep provides a large animal model that will enable studies of pathology and evaluation of therapies to treat this common lysosomal storage disorder.
Asunto(s)
Enfermedad de Gaucher/diagnóstico , Enfermedad de Gaucher/veterinaria , Enfermedades de las Ovejas/diagnóstico , Secuencia de Aminoácidos , Animales , Células Cultivadas , Análisis Mutacional de ADN , Enfermedad de Gaucher/patología , Glucosilceramidasa/análisis , Glucosilceramidasa/genética , Glucosilceramidasa/metabolismo , Homocigoto , Humanos , Datos de Secuencia Molecular , Mutación Missense , Homología de Secuencia de Aminoácido , Ovinos , Enfermedades de las Ovejas/etiología , Enfermedades de las Ovejas/patologíaRESUMEN
A herpes-like virus was for the first time purified from abalone diagnosed with ganglioneuritis. Pleuropedal ganglia, pedal nerve cords, head and epipodial tissue was collected and homogenized from abalone populations exhibiting high mortality and clinical signs consistent with herpes-virus like ganglioneuritis. Following ultracentrifugation by sucrose gradient prepared in sea-water, the purified virus was negatively stained and examined under a transmission electron microscope. Virus particles were observed to have an icosahedral capsid appearance surrounded by an envelope with numerous spikes on the external surface. The capsid ranged 92-109 nm in diameter and the enveloped virus was approximately 150 nm in diameter. Virus particles were found mainly at the interface of 40-50% sucrose gradients, and a few presented at the interface of 50-60% sucrose gradients. Isopycnic gradient centrifugation was performed in a potassium tartrate gradient and caesium chloride gradient, where the buoyant density of the herpes-like virus was determined to be 1.17-1.18 g/mL. The use of sea-water as the buffer in preparation of the gradient was critical in the preliminary purification of the herpes-like virus, and more efficient harvesting of the virus was achieved by sucrose and potassium tartrate gradients than caesium chloride gradient. The described method, whilst proving successful for purifying a herpes-like virus from abalone, may also be applicable to other viruses from marine animals.
Asunto(s)
Gastrópodos/virología , Herpesviridae/aislamiento & purificación , Neuritis/virología , Animales , Cápside/ultraestructura , Centrifugación por Gradiente de Densidad , Herpesviridae/ultraestructura , Microscopía Electrónica de Transmisión , Agua de Mar , Coloración y EtiquetadoRESUMEN
Iridoviruses have emerged over 20 years to cause epizootics in finfish and amphibians in many countries. They may have originated in tropical Asia and spread through trade in farmed food fish or ornamental fish, but this has been difficult to prove. Consequently, MCP, ATPase and other viral genes were sequenced from archival formalin-fixed, paraffin-embedded tissues from farmed Murray cod (Maccullochella peelii peelii) that died during an epizootic in 2003 and from diseased gouramis that had been imported from Asia. There was almost complete homology (99.95%) over 4,527 bp between Murray cod iridovirus (MCIV) and an iridovirus (DGIV) present in dwarf gourami (Colisa lalia) that had died in aquarium shops in Australia in 2004, and very high homology with infectious spleen and kidney necrosis virus (ISKNV) (99.9%). These viruses are most likely to be a single species within the genus Megalocytivirus and probably have a common geographic origin. Primers for genus-specific PCR and for rapid discrimination of MCIV/DGIV/ISKNV and red sea bream iridovirus (RSIV), a notifiable pathogen, were developed. These were used in a survey to determine that the prevalence of DGIV infection in diseased gourami in retail aquarium shops in Sydney was 22% (95% confidence limits 15-31%). The global trade in ornamental fish may facilitate the spread of Megalocytivirus and enable emergence of disease in new host species in distant biogeographic regions.