Transmission of Bluetongue Virus Serotype 8 by Artificial Insemination with Frozen-Thawed Semen from Naturally Infected Bulls

Abstract

Transmission of bluetongue (BT) virus serotype 8 (BTV-8) via artificial insemination of contaminated frozen semen from naturally infected bulls was investigated in two independent experiments. Healthy, BT negative heifers were hormonally synchronized and artificially inseminated at oestrus. In total, six groups of three heifers received semen from four batches derived from three bulls naturally infected with BTV-8. Each experiment included one control heifer that was not inseminated and that remained BT negative throughout. BTV viraemia and seroconversion were determined in 8 out of 18 inseminated heifers, and BTV was isolated from five of these animals. These eight heifers only displayed mild clinical signs of BT, if any at all, but six of them experienced pregnancy loss between weeks four and eight of gestation, and five of them became BT PCR and antibody positive. The other two infected heifers gave birth at term to two healthy and BT negative calves. The BT viral load varied among the semen batches used and this had a significant impact on the infection rate, the time of onset of viraemia post artificial insemination, and the gestational stage at which pregnancy loss occurred. These results, which confirm unusual features of BTV-8 infection, should not be extrapolated to infection with other BTV strains without thorough evaluation. This study also adds weight to the hypothesis that the re-emergence of BTV-8 in France in 2015 may be attributable to the use of contaminated bovine semen.

Keywords: BTV-8; artificial insemination; bluetongue; bluetongue virus; cattle; semen; serotype 8; transmission.

Figure 1

RNAemia as determined by real-time RT-PCR detection of BTV genome in blood of heifers infected with BTV-8 contaminated semen from natural-infected bulls. dpai: days post artificial insemination.

Figure 2

The level of antibodies in blood of heifers infected with BTV-8 contaminated semen from natural-infected bulls, as determined by a competitive ELISA and expressed in percentage negativity (PN). dpai: days post artificial insemination.

Figure 3

The body temperature of heifers infected with BTV-8 contaminated semen from natural-infected bulls until 35 days post-artificial insemination (dpai); thereafter, temperature remained normal.