Studies of Genetic Relationships between Bovine, Caprine, Cervine, and Rangiferine Alphaherpesviruses and Improved Molecular Methods for Virus Detection and Identification

Abstract

The glycoprotein B (gB) and D (gD) genes from five ruminant alphaherpesviruses, bovine herpesvirus 1 (BHV-1), bovine herpesvirus 5 (BHV-5), caprine herpesvirus 1 (CapHV-1), cervine herpesvirus 1, and rangiferine herpesvirus 1, were partially sequenced. The nucleotide sequence alignments revealed a highly conserved gB gene, with homologies ranging between 87.2 and 99.6%, and a more variable gD gene, with homologies ranging between 71.3 and 98.9%. The phylogenetic analysis of the gB and gD nucleotide and deduced amino acid sequences revealed that BHV-5 is the most closely related virus to the BHV-1 subtype 1 and BHV-1 subtype 2 cluster and that CapHV-1 is the most distantly related virus. The phylogenetic data showed a close relationship of all the studied viruses with suid herpesvirus 1. On the basis of sequence data for the gB gene, a nested PCR combined with restriction enzyme analysis (REA) of the PCR products was developed for the simultaneous detection and identification of the viruses that were studied. Nested primers from highly conserved sequence stretches were selected in order to amplify a region of 294 bp in all five viruses, and a subsequent REA of the PCR products allowed specific identification. A mimic molecule that served as an internal standard of the amplification efficiency was constructed. The practical diagnostic applicability of the assay was evaluated with clinical samples consisting of semen and organ specimens from experimentally infected animals.

Evidence of parapox-, alphaherpes- and pestivirus infections in carcasses of semi-domesticated reindeer (Rangifer tarandus tarandus) from Finnmark, Norway

“…In contrast to the situation for parapoxvirus, which seems to be transferred from domestic animals (sheep and goats) to reindeer, there are several factors that suggest that reindeer are hosting their own alphaherpes-and pestivirus variants. Herpesvirus has been isolated from reindeer in both Finland (Ek-Kommonen et al, 1986) and Sweden (Rockborn et al, 1990) and due to characterisations by means of serology (Lyaku et al, 1992), genomic restriction fragment length polymorphism (RFLP; Rimstad et al, 1993) and PCR and RFLP on amplicons from the glycoprotein-B and -D genes (Lyaku et al, 1996;Ros & Belák, 1999), it seems that reindeer herpesvirus (RanHV-1) is a virus variant closely related to herpesvirus in deer (cervid herpesvirus; CerHV-1) and goats (caprine herpesvirus; CapHV-1). Also the fact that the disease BVD in cattle are about to be eradicated and that the syndromes caused by herpesvirus in cattle (infectious bovine rhinotracheitis and infectious pustular vulvovaginitis and balanoposthitis) have not been registered in cattle in Norway since 1993 (Nyberg et al, 2003), are strong indications of the existence of distinct herpes-and pestivirus species in reindeer.…”

Effects of Bovine Herpesvirus Type 1 Infection in Calves with Maternal Antibodies on Immune Response and Virus Latency

“…Currently, there is no means to diagnose a latent infection other than the dexamethasone treatment (through concurrent serological and virological testing) (33), which is difficult to perform routinely. On the other hand, PCR examination of trigeminal ganglion can be applied only to a dead animal (35).…”

Evaluation of total oxidative stress and total antioxidant status in cows with natural bovine herpesvirus-1 infection

“…The anticoagulant-treated blood and nasal swab samples were centrifuged at 800 × g for 10 min at 4°C and the 2 mL of supernatants were collected as described elsewhere (Ataseven et al, 2009). These samples were used to confirm BHV-1 infection by DNA detection, using nested PCR as described elsewhere (Ros and Belak, 1999). The serum samples were also collected into clotactivator tubes (Ayset, Adana, Turkey) and then used for TAS, TOS, and other biochemical variables.…”

“…The QIAamp MinElute Virus Spin kit (Qiagen, Hilden, Germany) was used to extract total DNA from 200-µL volumes of the leukocyte and nasal swab samples. A 294-base pair conserved region of the glycoprotein B gene of BHV-1 was amplified by PCR, using first and second round primers as described by Ros and Belak (1999). The PCR reactions were performed using a thermal cycler (Techne 3000G, Bibby Scientific Ltd., Staffordshire, UK).…”