Isolation and Molecular Identification of BHV-1 From Cattle Suffering From Respiratory Signs

Abstract

A total of 93 ocular, 50 nasal swabs and 10 lung tissue were collected from 2-3 years old cattles affected with respiratory manifestation, and 145 serum samples from in contact apparently healthy cattles distributed in 4 different Governorates of Egypt (Ismalia, Qalioubia, Kafr El-Sheikh, and EL-Fayoum). Swabs and lung tissue samples were subjected to Bovine Herpes Virus (BHV-1) isolation on tissue culture Medin-Darby Bovine Kidney (MDBK) cell line and isolates were identified by direct fluorescent antibody technique (FAT) and polymerase chain reaction (PCR) by using primers complementary to the sequence of BHV-1 glycoprotein gB gene, as a rapid and more sensitive technique. From the results of virus isolation and identification were confirmed the more sensitivity of virus isolation than FAT and the most sensitivity and rapidity of PCR technique with confirmation and agreement of its results with the conventional methods. BHV-1 specific antibodies was detected in serum samples collected from the same 4 different Governorate, because of virus latency is a normal sequel to BHV-1 infection, by using the most suitable, sensitive, rapid, and specific techniques are ELISA and serum neutralization test (SNT), it is clear that the highest percentage of positive BHV-1 serum reactors was, in Kafr El-Sheikh followed by Ismalia, and Qalioubia and the lowest percentage was in EL-Fayoum. By the detection and identification of BHV-1 in collected samples and presence of positive reactors in serum samples this was a good indication for the circulation of BHV-1 in cattles of Egypt.

Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey

“…In the present study, BRSV isolation was attempted using samples from the clinically ill animals; however, BRSV could not be isolated from any samples using MDBK cells even though three blind passages were made. Similar results have been obtained in other studies, where no CPE was seen in cell cultures used for virus isolation [ 73 , 74 ]. Yazıci and others [ 37 ] reported CPE after 72 h of incubation in MDBK cell cultures, whereas Arns and others [ 72 ] were able to detect CPE in MDBK cells in only 1 of 33 samples after nine blind passages.…”

“…The limitation of virus isolation is the low number of repassages, as three passages seem to not be enough. However, as discussed above, both three repassages and higher numbers of repassages have been used in other studies [ 72 , 73 , 74 ]. Another limitation of our study is that we could not differentiate vaccinated from infected animals, as DIVA BRSV vaccines are not available yet.…”