The immune response to a Coxiella burnetii vaccine in sheep varies according to their natural pre-exposure.

Q fever in humans is caused by Coxiella (C.) burnetii. In 2008 and 2012, cases of Q fever in humans were linked to an infected flock of approximately 650 ewes. Since 2013 gimmers (G'13, G'14, G'15 etc.) were primary vaccinated (two doses) with an inactivated C.burnetii vaccine without any revaccination. In 2013, 30 ewes were primary vaccinated (A'13). Shedding was annually monitored by qPCR-testing of vaginal and nasal swabs collected at lambing. Animals were tested for Phase I- (PhI) and PhII-antibodies (Ab) and for PhII-specific-interferon-γ (IFN-γ) before and after vaccination. The effect of a revaccination was determined in 2018 and 2023. Groups of randomly selected gimmers primary vaccinated in 2015, 2016 and 2017 and a mixed group of older animals (A'13, G'13 and G'14) were revaccinated once in 2018. The trial was repeated in 2023 on groups primary vaccinated in 2019-2023. Major shedding after the outbreak in 2012 ceased in 2014. Thereafter C.burnetii was only sporadically detected at low-level in 2018, 2021 and 2023. Sheep naturally exposed to C.burnetii during the outbreak in 2012 (A'13, G'13) mounted a strong and complete (PhI, PhII, IFN-γ) recall immune response after vaccination. A serological PhI+/PhII+ pattern dominated after vaccination. In contrast, since 2014 a weaker immune response (PhII-titre, IFN-γ) and a dominance of the PhI-/PhII+ pattern was observed in vaccinated gimmers. The number of serologically non-responding gimmers to vaccination increased to 25.0 % in G'16/G'17 and 40.4 % in G'19/G'20. But revaccination even three (G'15 in 2018) and four (G'19 in 2023) years after primary vaccination resulted in a strong and complete immune response. No difference of the immune response nor to more recently primary vaccinated animals (G'23 in 2023) nor to those animals that were present during the outbreak (A'13/G'13/G'14 in 2018) was observed.

Long-term control of Coxiellosis in sheep by annual primary vaccination of gimmers.

Coxiella (C.) burnetii, a Gram-negative intracellular bacterium, causes Q fever in humans and Coxiellosis in animals. Ruminants are a primary source of human infection with C.burnetii. In 2013, vaccination was implemented in a sheep flock with 650 ewes associated with two outbreaks of Q fever in humans in 2008 and 2012. Only gimmers (yearlings) received two doses of a commercial C.burnetii phase I whole cell vaccine three weeks apart (primary vaccination) without any revaccination. Vaginal and nasal swabs collected shortly after lambing were tested by qPCR. Additionally, a group of non-vaccinated sentinels was serologically monitored for phase I (PhI), II (PhII) antibodies and for Interferon γ (IFN-γ) after stimulation of whole blood cells with PhII-antigen with and without an IL-10-neutralizing monoclonal antibody. In 2021, 679 sera collected in 2014-2021 were retested retrospectively with three commercial ELISA kits and one batch of an in-house PhI/PhII-ELISA. A low-level shedding of C.burnetii (<103 mean C.burnetii/swab) was observed until 2014. In 2021 C.burnetii was detected in two animals (<103.1C.burnetii/swab), but vaginal swabs collected at two subsequent lambing seasons remained negative. Seroconversion of sentinels was detected until 2017. However, the retrospective analysis of sentinels in 2021 revealed additional single seropositive animals from 2018 to 2021. IFN-γ reactivity was observed during the whole study period; it peaked in 2014 and in 2018 and decreased thereafter. The sporadic detection of C.burnetii and the immune responses of sentinels suggested that a subliminal infection persisted despite vaccination. Nevertheless, vaccination of gimmers prevented the development of a major outbreak, it controlled the infection and reduced the risk of human infection.

Insights into the dynamics of endemic Coxiella burnetii infection in cattle by application of phase-specific ELISAs in an infected dairy herd.

Serological diagnosis of acute and chronic Q fever in humans relies on detection of antibodies to phase I (PhI) and II (PhII) antigens of Coxiella (C.) burnetii. Although phase-specific antigens are available, they are not yet used in ruminants as they are in humans. This study focuses on phase-specific serology as a tool for analysis of the dynamics of infection in cattle. As a prerequisite, sero-prevalence in Bavarian cattle (1) and sero-prevalences for age-groups (2) were determined by ELISA (CHEKIT Q-Fever; mix of PhI/PhII-antigen). Subsequently, phase-specific antigens were coated onto ELISA plates individually and tests were simultaneously applied in an endemically infected herd with about 90 dairy cows and 250 calves/heifers in April 2005, March 2006 and retrospectively in May and October 2004. From April 2005 onward, placentas were analysed for C. burnetii by PCR (3). (1) Sero- and herd prevalences based on 21,051 sera from 603 Bavarian dairy farms collected in 2003 were 14.8% ± 0.48% and 72.3% ± 3.6%, respectively. (2) Analysis of 3965 animals from 105 farms for which age was reported revealed a base level of sero-prevalence of less than 5% in 1-2 years old animals, it increased to 15% in 2-3 years old and reached a plateau (25-30%) in cows four years and older. (3) In May 2004 and April 2005 a peak of PhI(-)/PhII(+)-prevalence in primiparous cows (2.0-3.5 years) was observed; but not in October 2004 and March 2006. The PhI(-)/PhII(+)-pattern in primiparous cows changed to negative (one-third), PhI(+)/PhII(+) (1/3) or persisted (1/3). In contrast, sero-conversion was rare in multiparous cows (>3.5 years). If the PhI(-)/PhII(+) pattern was detected, it was due to an infection in preceding years. This pattern persisted (2/3) or changed to negative (1/3); a change to PhI(+)/PhII(+) did not occur. PhI(-)/PhII(+) in heifers (1-2 years) always changed to negative. Detection of PhII-antibodies was significantly associated with PCR-positive placentas. Remarkably, 45% of sera with the PhI(-)/PhII(+) pattern were negative for the CHEKIT Q-Fever ELISA, thus this test missed an important group of infected animals.