ABSTRACT
Malignant catarrhal fever (MCF), a fatal viral disease of cattle and other large ruminants, has a worldwide distribution. There are two forms of the disease, one of which, is caused by Alcelaphine herpesvirus-1 (AHV-1) and is derived from wildebeest. The other form is associated with domestic sheep and is caused by ovine herpesvirus-2 (OHV-2). The disease in Indonesia is sheep-associated with the preferred livestock of this area, Balinese cattle (Bos javanicus) and water buffalo (Bubalus bubalis), both highly susceptible to SA-MCF. The incidence in these species is thought to be high but the prevalence and economic losses attributable to SA-MCF have been difficult to assess. a polymerase chain reaction (PCR) test, based on a cloned OHV-2 gene sequence, was successfully applied to the detection of OHV-2 DNA in normal sheep and animals affected with SA-MCF. OHV-2 DNA was detected in eleven confirmed cases of SA-MCF and in the peripheral blood leucocyte (PBL) fraction of six latently infected sheep. These findings have confirmed that the PCR can be of value in establishing a diagnosis of MCF and that the aetiological agent of MCF in Indonesia is OHV-2. The amplification of DNA from the PBL of goats suggests that they are infected with a similar or identical herpesvirus.
Subject(s)
DNA, Viral/analysis , Herpesviridae/isolation & purification , Malignant Catarrh/virology , Ruminants/virology , Animals , Blotting, Southern/veterinary , Buffaloes/virology , Cattle , Goats/virology , Indonesia , Malignant Catarrh/diagnosis , Polymerase Chain Reaction/veterinary , Sheep/virologyABSTRACT
From a genomic library previously constructed from a lymphoblastoid cell line (LCL) propagated from a bovine case of sheep-associated malignant catarrhal fever (SA-MCF), caused by ovine herpesvirus-2 (OHV-2), several OHV-2 clones were identified and characterised by hybridisation using probes from the unique region of the Alcelaphine herpesvirus-1 (AVH-1) genome. Nucleotide sequence from one clone was generated and the predicted amino acid sequence was found to contain regions of homology with the 140 and 160 kDa tegument proteins of Epstein-Barr virus and herpesvirus saimiri respectively. Oligonucleotide primers were constructed and a polymerase chain reaction (PCR) test was developed for the detection of OHV-2 viral DNA. Amplified product was identified by restriction with RsaI and BmyI. The primers were highly specific for OHV-2 DNA with a limit of detection of 6.4 pg of genomic DNA derived from the parent LCL. This was estimated to correspond to one diploid bovine cell. The PCR was successfully applied to detect OHV-2 DNA in peripheral blood leucocytes (pbl) from clinical cases of SA-MCF and normal sheep.
Subject(s)
DNA, Viral/isolation & purification , Herpesviridae/isolation & purification , Malignant Catarrh/microbiology , Polymerase Chain Reaction/veterinary , Sheep Diseases/microbiology , Amino Acid Sequence , Animals , Base Sequence , Cattle , Cell Line , Cells, Cultured , DNA, Single-Stranded , Deer , Herpesviridae/classification , Herpesviridae/genetics , Molecular Sequence Data , Rabbits , Sensitivity and Specificity , Sequence Homology, Amino Acid , SheepABSTRACT
A polymerase chain reaction test for the detection of ovine herpesvirus-2 (OHV-2) DNA was used to identify sites of OHV-2 infection in peri-natal lambs and in adult sheep. OHV-2 was detected in the nasal secretions from all lambs within a period of two months following birth. Subsequently, OHV-2 DNA was identified in a number of epithelial tissues including the cornea, turbinates and pharynx. In addition, OHV-2 DNA was detected exclusively in B-lymphocytes from six of ten adult sheep tested. An infection cycle for OHV-2 in sheep is proposed which bears similarities with the gammaherpesviruses Epstein-Barr virus and mouse herpesvirus-68.
Subject(s)
Herpesviridae Infections/diagnosis , Herpesviridae/isolation & purification , Sheep Diseases/diagnosis , Animals , B-Lymphocytes/virology , DNA, Viral , Herpesviridae/genetics , Herpesviridae Infections/veterinary , Herpesviridae Infections/virology , Nasal Mucosa/metabolism , Nasal Mucosa/virology , Nucleic Acid Hybridization , Sheep , Sheep Diseases/virologyABSTRACT
Analysis of the genomic DNAs of chlamydial isolates from sheep, cattle, and pigs was performed by Southern blot hybridization and by restriction endonuclease (RE) profiling of DNA amplified by PCR. The hybridization probes were derived from whole genomic DNA, the major outer membrane protein (MOMP) gene, the 16S rRNA gene, and an avian Chlamydia psittaci isolate plasmid. The PCR analysis used targets in the MOMP gene, the 16S rRNA gene, and the 60-kDa cysteine-rich protein gene. Together, the results showed that although there was considerable heterogeneity in the DNA sequence in the MOMP gene region, all the isolates had the same underlying total genomic RE profiles and yielded identical RE profiles for the rRNA and 60-kDa-protein gene regions. Most of the isolates were found to hybridize with the plasmid probe. Comparison of the MOMP sequence of one of the isolates (P787) with that of a known Chlamydia pecorum strain together with the results of the RE analyses allowed the conclusion that the isolates should all be classified within this new species.