During summer 2011, cattle presented severe hyperthermia combined with dropped milk yield and diarrhoea from unknown origin. In October 2011, blood was collected from cattle presenting these clinical signs in Schmallenberg, a small city in West Germany. A new Orthobunyavirus, responsible for these unspecific clinical signs was identified and named Schmallenberg virus (SBV). Upon November 2011, an epizootic outbreak of abortion, stillbirths and malformed new-born was observed in bovine, ovine and caprine herds in Europe due to transplacental transmission of SBV to the foetus. The SBV vectors are small hematophagous midges of the gender Culicoides. This work contributed to estimate the impact of the SBV epidemic in Belgium (Study 1). On the basis of farmer’s observations, between 0.5% and 4% of calves were aborted, stillborn or malformed due to SBV in 2011-2012. Abortions and stillbirths were not clear consequences of the SBV outbreak in cattle. In sheep, between 11% and 19% of lambs were aborted, stillborn or malformed due to SBV in 2011-2012. Deformed animal was the most important finding of SBV outbreak at herd level and an essential condition for the farmer to send suspected samples to the National Reference Laboratory (NRL) for SBV analysis. The results gathered from the study indicate that SBV surveillance and monitoring should be implemented by SBV RNA detection with rRT-PCR in organs collected from stillborn and deformed calves and lambs born in big herds. The high impact of SBV highlighted in the Study 1 was putatively explained by unknown host supporting the SBV activity. In this respect, the role of pigs had never been evaluated. This was essential considering the suggested role of the domestic pigs in the life-cycle of the SBV-closely related Akabane virus (AKAV) (Huang et al., 2003). The absence of RNAemia after experimental infection of piglets with SBV realized in the Study 2 of the thesis suggests the absence of obvious role of domestic pigs in SBV life-cycle. The absence of RNAemia is indeed a strong indication that further spread of SBV from the pigs to the Culicoides during a blood meal of the vector is not likely to occur, therefore making impossible an SBV transmission. The limited and temporary seroconversion observed after SBV inoculation in only half of the inoculated piglets and the absence of seroconversion reported in a limited number of field collected samples support this consideration. To prevent SBV progression, it was crucial to further study the pathogenesis of SBV. The Study 1 proved that the most important clinical impact of SBV was the consequence of the malformed new-born; hereto it was particularly crucial to improve the knowledge on the development of the SBV-related teratogenic effects. In this respect, experimental infection of pregnant sheep with SBV constituted an appropriate research approach. An experimental model was therefore essential to standardize. This thesis contributed to the standardization of in vivo experiments (in collaboration with another working group) by determining the minimum infectious dose of an SBV infectious inoculum. This reference infectious serum must contain approximately 20 TCID50 to induce a homogeneous effective infection in sheep. This dose is rather low and could be inoculated by a single Culicoides under natural conditions. Beyond this minimum infectious dose, no dose dependent effect was observed in productively inoculated ewes, either in the duration of the RNAemia, the quantity of SBV RNA detected by rRT–PCR in the blood, or in the number of SBV RNA copies present in the organs collected at necropsy. The experimental model developed (partly) in the Study 3 was used to inoculate pregnant ewes at day 45 and 60 of gestation, and increase the knowledge on SBV transplacental transmission. The inoculation induced the persistence of SBV RNA in placental organs until birth. Schmallenberg virus RNA was recovered from the organs collected at birth from the lambs of both groups. However, the chance to obtain SBV RNA positive placental organs was significantly higher when the infectious inoculum was inoculated at day 60 of gestation. Positive organs in lambs included CNS and muscle, but no malformation was observed in new-born lambs. This absence of malformations suggests that SBV inoculation must occur earlier than the day 45 of gestation to produce teratogenic effects in sheep. Also, the persistence of SBV RNA in the foetal envelope is indicative of a putative mean for SBV overwintering. The Study 4 highlighted a 6 month persistent seroconversion in the absence of SBV surinfection. In the meantime, SBV circulation drastically dropped on the field and the absence of SBV circulation could induce the sheep to become seronegative under natural conditions. In the Study 5, the experimental model developed in the Study 3 was used to demonstrate that one single SBV inoculation can induce a protective immunity in sheep that persists during a minimum period of 15 months. This experiment highlights that 2 successive periods of SBV circulation, spared of one year, is not likely to induce malformations on the field the second year. Based on the experience gathered with the closely related AKAV, recurrent outbreaks of congenital events can be expected for a long period. Vaccination of seronegative animals could be used to prevent the deleterious effects of SBV in case of SBV re-emergence. During this epidemic, different surveillance approaches including syndromic surveillance, sentinel herd surveillance, cross-sectional seroprevalence studies and pathogen surveillance in vectors have proven their utility and complementarity and should be considered to continue in the future in order to monitor the SBV dynamic.
9 Dec 2015
Examination held at
Faculty of Veterinary Medicine, University of Liège