Progression of Coxiella burnetii infection after implementing a two-year vaccination program in a naturally infected dairy cattle herd

Background

The high prevalence of Coxiella burnetii infection in dairy cattle herds recently reported and the long survival time of the bacterium in the environment pose a risk to human and animal health that calls for the implementation of control measures at herd level. This study presents the results of a 2-year vaccination program with an inactivated phase I vaccine in a Spanish dairy herd naturally infected with C. burnetii. Calves older than 3 months and non-pregnant heifers and cows were vaccinated in April 2011 and the farm was subsequently visited at a monthly basis for vaccination of recently calved cows and calves that reached the age of 3 months. Annual booster doses were given to previous vaccinated animals as well. The effectiveness of the vaccine was assessed in terms of level of C. burnetii shedding through milk and uterine fluids and environmental contamination as determined by polymerase chain reaction (PCR).

Results

The percentage of shedder animals through uterine fluids and milk progressively decreased, and C. burnetii DNA load in bulk-tank milk samples was low at the end of the study. The average seroconversion rate in not yet vaccinated animals, which acted as control group, was 8.6% during the first year and 0% in the second year. DNA of C. burnetii was found in aerosols and dust samples taken in the calving area only at the beginning of the study, whereas slurry samples remained C. burnetii PCR positive for at least 18 months. Multiple Locus Variable number tandem-repeat Analysis identified the same genotype in all C. burnetii DNA positive samples.

Conclusions

In the absence of any changes in biosecurity, the overall reduction of C. burnetii infection in animals to 1.2% milk shedders and the reduced environment contamination found at the end of the study was ascribed to the effects of vaccination together with the culling of milk shedders. Vaccination has to be planned as a medium-long term strategy to suppress risks of re-infection.     

Q fever (Coxiella burnetii) investigations in dairy cattle: persistence of antibodies after vaccination.

The immune response and persistence of antibodies were investigated in dairy cattle vaccinated with formalin-inactivated phase (ph) I Coxiella burnetii vaccine agglutinating antibody geometric mean titer (GMT) of 193.2 at 1 month after vaccination compared to a GMT of 2.0 for nonvaccinated calves. The agglutinating antibodies gradually decreased in vaccinated cattle, but the GMT remained approximately 4 times higher than that for the nonvaccinated group for at least 20 months. Results of serotests at 2 months after revaccination indicated a rapid increase in the GMT to 177.0 with agglutinating titers between 1:64 and 1:512.     

Prevalence of Coxiella burnetii antibodies in Portuguese dairy cattle herds

Q fever is an important zoonotic disease which has been recently diagnosed, mainly in sheep and goats, in Portugal. The aim of the present study was to determine the prevalence of bovine Coxiella burnetii antibodies in dairy farms from the northwest of Portugal. Bulk tank milk samples were randomly obtained, on November 2013, from 90 dairy farms and assayed using an ELISA kit. The apparent prevalence was 61.1 % (95 % C.I. from 50.8 to 70.5 %). The proportion of negative and intermediate (inconclusive) herds was 34.5 % (25.5 to 44.7 %) and 4.4 % (1.7 to 10.9 %), respectively. In conclusion, a high level of exposure to Coxiella burnetii was observed in Portuguese dairy cattle herds, highlighting the needs to better understand the epidemiology of Q fever in Portugal by the implementation of a monitoring program based on harmonized serologic and molecular methodologies and elucidation of the infection status of the herds.

     

Coxiella burnetii infections and infections with bacteria of the genus Chlamydia in dairy cattle

Comparative studies on the prevalence of infections caused by Coxiella burnetii (C. burnetii) and Chlamydia were carried out with 592 cattle older than 2 years and 234 cattle younger than 2 years. Of these 477 originated from 24 dairy herds with considerable fertility problems (positive herds) and 349 from 14 dairy herds without major fertility problems (control herds). For the direct detection of these pathogens in the genitals capture ELISAs were employed, for the demonstration of antibodies the complement fixation test (CFT). Direct detection of C. burnetii and Chlamydia single as well as mixed infection revealed significant higher values for cattle from positive herds compared with those from the control herds. Animals revealing insemination ratios of > or = 2 showed significantly more frequent excretion of Chlamydia via the genitals and antibodies against C. burnetii than cattle with an insemination ratio of < 2. Investigations of cows which had had an abortion showed no indications of significantly more frequent C. burnetii or chlamydial infections. Inseminated but non-pregnant cows excreted significantly more C. burnetii and Chlamydia than pregnant cows. Clinical signs of endometritis were associated with an enhanced excretion of Chlamydia. Animals younger than 2 years excreted significantly more frequently C. burnetii but not Chlamydia via the genitals than animals older than 2 years. Indirect test showed results vice versa.     

Serological evidence of Coxiella burnetii infection in beef cattle in Queensland

Background Queensland has the highest incidence of Q fever in Australia. The aim of this study was to undertake a cross‐sectional seroprevalence survey of Coxiella burnetii, the causative agent of Q fever, in beef cattle in Queensland. Methods Serum samples were tested by ELISA for both phase II and phase I antigens of the organism using an Australian isolate. Blood samples were collected at an abattoir that processes beef cattle originating from northern and north‐western Queensland, in addition to blood samples taken from beef cattle across Queensland as part of a second survey. Results Seropositivity was 16.8% (95% confidence interval 16.7–16.8%). Conclusion Evidence of C. burnetii infection in beef cattle has public health implications for occupational exposure of primary producers and veterinarians and for the proximity of beef cattle properties to residential areas in regional Queensland. This study is the first known investigation of C. burnetii seroprevalence in beef cattle in Queensland and the first known use of an Australian C. burnetii isolate for screening using both phase II and phase I antigens.     

The incidence of Coxiella burnetii antibodies in cattle in the Transvaal

With the use of the complement fixation test, 8,900 cattle were tested for antibodies to Coxiella burnetii. These were randomly selected from 178 different farms in 37 districts in the Transvaal. The percentage of cattle in the sample with positive antibody titres was equal to 7.78%, with a standard error of 0.28%. Because of the large size of the sample, asymptotic normality can be relied upon and the population confidence interval calculated. This was found to be greater than or = 0.07 and less than or = 0.085 at a 99% confidence level. Hence we are 99% confident that between 7% and 8.5% of the cattle in the Transvaal had antibodies to Coxiella burnetii during the period March 1985 to July 1986. The proportion of cattle with C. burnetii antibodies was also estimated for each of the 37 districts tested. Every district tested had some evidence of C. burnetii. The percentage of positive titres ranged from less than 1%-30% per district. This suggests that C. burnetii is probably an endemic disease of the cattle population of the Transvaal. A higher proportion of cattle had antibody titres in the central and south-eastern parts of the Transvaal. This distribution may be linked to the distribution of Boophilus species ticks which occur in the same areas of the Transvaal.     

Increasing prevalence of Coxiella burnetii seropositive Danish dairy cattle herds

A study based on bulk tank milk samples from 120 randomly selected dairy cattle herds was conducted to estimate the prevalence of Coxiella burnetii seropositive dairy herds, to describe the geographical distribution, and to identify risk factors. Using the CHEKIT Q-fever Antibody ELISA Test Kit (IDEXX), the study revealed a prevalence of 79.2% seropositive herds, 18.3% seronegative herds, and 2.5% serointermediate herds based on the instructions provided by the manufacturer. Multifactorial logistic regression showed statistically significant associations (P < 0.01) between C. burnetii seropositivity and increasing herd size (OR = 1.02 per cow increment) and increasing regional average number of cattle per dairy herd (OR = 1.02 per animal increment). Herds >150 cows had 17.9 times higher odds of testing positive compared to herds <80 cows. The regional average number of cattle herds per square kilometer was borderline significantly related to the occurrence of seropositive dairy herds (P = 0.06). The results indicate an increased prevalence of seropositive dairy herds since the previous survey in 2008 and an adverse impact of increasing herd size and cattle density on the risk of seropositivity.     

Seroprevalence and risk factors of Coxiella burnetii infection in cattle in northeast Algeria

Tropical Animal Health and Production - A cross-sectional study was conducted to determine the seroprevalence and the risk factors associated with C. burnetii infection in cattle in the state of...     

Analysis of caprine IgG1 and IgG2 subclass responses to Chlamydia psittaci infection and vaccination

Enzyme-linked immunosorbent assays (ELISAs) specific for caprine IgG(H+L), IgG1 and IgG2 were developed and evaluated for serodiagnosis of Chlamydia psittaci infections in a Tunisian goat flock with currently occurring chlamydial abortions and a clinically inapparent goat flock of an animal research facility. Additionally, ELISAs were applied to record the IgG1 and IgG2 dynamics of four goats vaccinated with inactivated Chlamydia psittaci and Coxiella burnetii. For screening purposes, the IgG(H+L) ELISA proved to be superior to the complement fixation test because it detected a larger number of chlamydial abortions and was easier to perform and to interpret. Analysis of Chlamydia psittaci-specific IgG1 and IgG2 responses to naturally occurring infections by ELISA revealed high IgG1 levels associated with IgG2 in goats with current abortions, whereas clinically inapparent, but seropositive goats were characterized by significantly lower IgG1 levels only (P less than 0.001). Similarly, the four vaccinated goats responded initially with Chlamydia psittaci-specific IgG1, whereas second and third vaccinations induced (as in goats with chlamydial abortions) predominantly IgG1, but also IgG2. The results indicated that clinically inapparent chlamydial infection may be distinguished from overt disease by analysing specific IgG1 and IgG2 responses. Applying Coxiella burnetii- specific ELISAs on field samples, IgG1 alone could be detected in eight, IgG2 alone in one and IgG1 combined with IgG2 in nine goats. The coxiella-specific antibody response of the four vaccinated goats was--in contrast to the chlamydia-specific response--characterized by IgG2 dominance.     

Association between Coxiella burnetii shedding in milk and subclinical mastitis in dairy cattle

The objective of this research was to explore the potential association between Coxiella burnetii shedding in milk and chronic subclinical mastitis in dairy cattle. In two separate studies, we identified an association between PCR-based detection of C. burnetii in milk and chronic subclinical mastitis in lactating dairy cows. These studies were conducted in a commercial dairy herd where there was ongoing intensive monitoring of subclinical mastitis by aerobic bacteriology, but no prior knowledge or management of C. burnetii infections. In a case-control study, quarter level C. burnetii status determined by real-time quantitative PCR (RT-qPCR) was strongly associated with chronic subclinical mastitis as measured by milk somatic cell counts. In a subsequent cross sectional study, 147 (45%) of 325 lactating cows were positive for C. burnetii by RT-qPCR of composite milk samples. In a generalized linear model, accounting for the effect of covariates including aerobic intramammary infection status, C. burnetii PCR status was a significant predictor of linear somatic cell count score. In agreement with a small number of previous reports, this research provides evidence that there may be mammary gland specific manifestations of C. burnetii infections in dairy cattle.     

Immune responses of cattle following vaccination with living and non-living Babesia bovis antigens

Twenty-four yearling Hereford (Bos taurus) cattle were vaccinated against Babesia bovis using either live parasites or non-living antigens obtained from the supernatant of in vitro cultures. A single dose of live parasites was given subcutaneously, while the non-living supernatant antigen (NLSA) was combined with saponin and 2 doses given, 2 weeks apart. Following vaccination with live parasites, serum antibodies remained at high levels for 6 months, but the lymphocyte transformation response was low and lasted only 10-18 days. In contrast, NLSA vaccination was followed, after 21-28 days, by a peak of serum antibodies which then slowly declined. The lymphocyte transformation response in these animals was much higher and persisted for 6 months. Following heterologous challenge all unvaccinated cattle had severe reactions and required treatment to prevent death. Cattle vaccinated with live parasites had mild reactions with only 1 of the 12 requiring treatment. Cattle vaccinated with NLSA were only partially protected and 6 of the 12 required treatment.     

Dominance of Chlamydia psittaci-specific IgG2 subclass in the humoral immune responses of naturally and experimentally infected cattle

Indirect enzyme-linked immunosorbent assays were applied to differentiate Chlamydia (C.) psittaci-specific IgG1 and IgG2 levels in 143 individual serum samples from cattle with naturally occurring chlamydial infections and in 190 sequential serum samples from 26 experimentally infected pregnant cows, calves, and a bull. The mean IgG1:IgG2 ratio of naturally infected cattle was 1:4 indicating a significant (p less than 0.001) IgG2 dominance. Similar ratios were detected in the experimentally infected cattle. The dominance of IgG2 was independent of breed, sex, and age. Twenty-nine cattle had significant immunoglobulin levels to both C. psittaci and Coxiella (C.) burnetii simultaneously. The predominance of C. psittaci-specific IgG2, in contrast to the predominance of C. burnetti-specific IgG1 detected in these same individual serum samples under identical conditions, indicates that the ability to preferentially produce either IgG1 or IgG2 was not limited in these individual cattle. A transient yet significant IgG1 response was also developed in cows following chlamydia-induced abortions (immunotype 1) or in cattle infected with the polyarthritis-serositis-encephalomyelitis agents (immunotype 2). IgG1 levels decreased faster than IgG2 levels. These findings have diagnostic implications and identify the need for determining the immunoglobulin classes and subclasses of the humoral immune responses of animals and man to chlamydial infections.     

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.     

Surveys on Coxiella burnetii infections in Swedish cattle,sheep, goats and moose

Background

Q fever is a zoonotic disease caused by the bacterium Coxiella burnetii. Prevalence data in ruminant species are important to support risk assessments regarding public and animal health. The aim was to investigate the presence of or exposure to C. burnetii in cattle, sheep, goats and moose, and to compare two enzyme-linked immunosorbent assays (ELISAs). National surveys of antibodies against C. burnetii were performed for dairy cattle (n=1537), dairy goats (n=58) and sheep (n=518). Bovine samples consisted of bulk milk, caprine of pooled milk, and ovine of pooled serum. Antibodies were investigated in moose samples (n=99) from three regions. A one-year regional cattle bulk milk survey was performed on the Isle of Gotland (n=119, four occasions). Cattle, sheep and goat samples were analysed with indirect ELISA and moose samples with complement fixation test. For the sheep, goat, and parts of the cattle survey, samples were run in parallel by ELISAs based on antigens from infected ruminants and ticks. Bulk milk samples from the regional cattle survey and vaginal swabs from a subset of the sheep herds (n=80) were analysed for the agent by polymerase chain reaction. Spatial clustering was investigated in the national cattle survey.

Results

The prevalence of antibodies in dairy herds was 8.2% with large regional differences. High risk clusters were identified in the southern regions. The prevalence among dairy herds on the Isle of Gotland varied from 55.9% to 64.6% and 46.4% to 58.9.0% for antibodies and agent, respectively, overall agreement between agent and antibodies was 85.2%. The prevalence of antibodies in sheep was 0.6%, the agent was not detected the vaginal swabs. Antibodies were not detected in goats or moose, although parts of the moose samples were collected in an area with high prevalence in cattle. The overall agreement between the two ELISAs was 90.4%.

Conclusions

The prevalence of antibodies against C. burnetii in dairy cattle in Sweden shows large regional differences. The results suggest that C. burnetii is a rare pathogen among Swedish moose, dairy goat and sheep. ELISAs based on ruminant and tick antigen performed in a similar manner under Swedish conditions.     

Effectiveness of vaccination and antibiotics to control Coxiella burnetii shedding around calving in dairy cows

Effectiveness of phase 1 vaccine, combined or not with tetracycline, to control Coxiella burnetii vaginal shedding at calving in cows was assessed through a 13 months study in 22 Q fever clinically affected commercial dairy herds. Four medical strategies implemented at herd level but randomly assigned to cows (vaccination, vaccination and tetracycline, tetracycline, nothing) were compared. There was no significant interaction effect between vaccination and antibiotherapy. Tetracycline used once at drying off was associated with a lower risk of being detected shedder at calving (OR=0.40, CI 95% [0.21-0.75]), but had no significant effect on the bacterial load shed. Vaccination did not significantly prevent shedding but was significantly (OR=0.15, CI 95% [0.03-0.85]) associated with lower bacterial load shed. Thus, vaccination using a phase 1 vaccine and antibiotherapy using tetracycline is associated with a decrease in shedding in dairy cows and could contribute to reduce the bacterial load generated in the environment. To the best of our knowledge, this is the first study providing useful information for evidenced-based and rational use of medical strategy combining antibiotic and vaccination in infected dairy cattle herds.