The 1B vaccine strain of Chlamydia abortus produces placental pathology indistinguishable from a wild type infection.

Chlamydia abortus is one of the most commonly diagnosed causes of infectious abortion in small ruminants worldwide. Control of the disease (Enzootic Abortion of Ewes or EAE) is achieved using the commercial live, attenuated C. abortus 1B vaccine strain, which can be distinguished from virulent wild-type (wt) strains by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Published studies applying this typing method and whole-genome sequence analyses to cases of EAE in vaccinated and non-vaccinated animals have provided strong evidence that the 1B strain is not attenuated and can infect the placenta causing disease in some ewes. Therefore, the objective of this study was to characterise the lesions found in the placentas of ewes vaccinated with the 1B strain and to compare these to those resulting from a wt infection. A C. abortus-free flock of multiparous adult ewes was vaccinated twice, over three breeding seasons, each before mating, with the commercial C. abortus 1B vaccine strain (Cevac® Chlamydia, Ceva Animal Health Ltd.). In the second lambing season following vaccination, placentas (n = 117) were collected at parturition and analysed by C. abortus-specific real-time quantitative PCR (qPCR). Two placentas, from a single ewe, which gave birth to live twin lambs, were found to be positive by qPCR and viable organisms were recovered and identified as vaccine type (vt) by PCR-RFLP, with no evidence of any wt strain being present. All cotyledons from the vt-infected placentas were analysed by histopathology and immunohistochemistry and compared to those from wt-infected placentas. Both vt-infected placentas showed lesions typical of those found in a wt infection in terms of their severity, distribution, and associated intensity of antigen labelling. These results conclusively demonstrate that the 1B strain can infect the placenta, producing typical EAE placental lesions that are indistinguishable from those found in wt infected animals.

Polymicrobial Infections In Brain Tissue From Alzheimer's Disease Patients.

Several studies have advanced the idea that the etiology of Alzheimer's disease (AD) could be microbial in origin. In the present study, we tested the possibility that polymicrobial infections exist in tissue from the entorhinal cortex/hippocampus region of patients with AD using immunohistochemistry (confocal laser scanning microscopy) and highly sensitive (nested) PCR. We found no evidence for expression of early (ICP0) or late (ICP5) proteins of herpes simplex virus type 1 (HSV-1) in brain sections. A polyclonal antibody against Borrelia detected structures that appeared not related to spirochetes, but rather to fungi. These structures were not found with a monoclonal antibody. Also, Borrelia DNA was undetectable by nested PCR in the ten patients analyzed. By contrast, two independent Chlamydophila antibodies revealed several structures that resembled fungal cells and hyphae, and prokaryotic cells, but most probably were unrelated to Chlamydophila spp. Finally, several structures that could belong to fungi or prokaryotes were detected using peptidoglycan and Clostridium antibodies, and PCR analysis revealed the presence of several bacteria in frozen brain tissue from AD patients. Thus, our results show that polymicrobial infections consisting of fungi and bacteria can be revealed in brain tissue from AD patients.

Serosurvey of Leptospira interrogans, Brucella abortus and Chlamydophila abortus infection in free-ranging giant anteaters (Myrmecophaga tridactyla) from Brazil / Inquérito sorológico de Leptospira interrogans, Brucella abortus e Chlamydophila abortus em tamanduás-bandeira (Myrmecophaga tridactyla) de vida livre no Brasil

A serological survey for antibodies against Leptospira interrogans, Brucella abortus, and Chlamydophila abortus was conducted in 21 clinically healthy, free-ranging giant ant- eaters (Myrmecophaga tridactyla) from Parque Nacional das Emas (Goiás State, Brazil; n=6), Parque Nacional da Serra da Canastra (Minas Gerais State, Brazil; n=9), and RPPN SESC Pantanal (Mato Grosso State, Brazil; n=6) between July 2001 and September 2006. Sera were screened for antibodies against 22 serovars of Leptospira interrogans with a microscopic agglutination test. Twelve tested positive for L. interrogansserovars sentot (n=5 in PN Emas, n=2 in PN Serra da Canastra), butembo (n=2 in PN Serra da Canastra), autumnalis, bataviae, and shermani/icterohaemorrhagiae(n=1 each in SESC Pantanal)One adult female tested positive for B. abortus with the buffered plate antigen test. All sera were negative for C. abortususing the complement fixation text. This is the first report of pathogens that may interfere with the reproduction and population dynamics of free-ranging giant anteaters.

Inquéritos sorológicos para detecção de anticorpos contra Leptospira interrogans, Brucella abortus, e Chlamydophila abortus foram realizados em 21 tamanduás-bandeira (Myrmecophaga tridactyla) de vida livre do Parque Nacional das Emas (Goiás, Brasil, n=6), o Parque Nacional da Serra da Canastra (Minas Gerais, Brasil, n=9) e RPPN SESC Pantanal (Mato Grosso, Brasil, n=6) entre julho de 2001 e setembro de 2006. Os sor os foram testados para anticorpos contra 22 sorotipos de Leptospira interrogans com um teste de aglutinação microscópica. Doze animais foram considerados positivos para L. interrogans sorovares sentot (n=5 em PN Emas, n=2 em PN Serra da Canastra), butembo (n=2 em PN Serra da Canastra), autumnalis, bataviae e shermani/icterohaemorrhagiae(n=1 para cada sorovar em SESC Pantanal). Uma fêmea adulta testou positivo para B. abortuscom o teste do antígeno tamponado. Todos os soros se mostraram negativos para C. abortusatravés do teste de fixação do complemento. Este é o primeiro relato de patógenos que podem interferir na dinâmica reprodutiva de populações de tamanduás em estado selvagem.

Seroprevalence and risk factors associated with Chlamydophila spp. infection in ewes in the northeast of Algeria.

A cross-sectional study was carried out to estimate prevalence of Chlamydophila spp. antibodies and to investigate risk factors associated with chlamydial infection in 552 ewes between March 2011 and January 2012 in the province of Constantine. Anti-Chlamydophila antibodies were detected using an indirect enzyme-linked immunosorbent assay kit in 24.5% of examined sera. Of the herds, 70.4% had at least one seropositive animal. A pretested structured questionnaire was administered in order to collect information on individual animal health and herd management practices. Chi-square analysis and multivariable logistic regression model were used to identify risk factors related to Chlamydophila seropositivity. Univariable analysis revealed 17 variables with p < 0.25 that were offered to the multivariable logistic regression model which in turn identified 12-23 months age group (OR = 5.903, 95% CI (OR) = 1.690; 20.618) and not using disinfectants (OR = 2.099, 95% CI (OR) = 1.314; 8.065) as risk factors for Chlamydophila spp. seropositivity. Moreover, occurrence of stillbirth problem (OR = 3.682, 95% CI (OR) = 1.825; 7.430) and 5-10% mortality rate in young lambs (OR = 2.584, 95% CI (OR) = 1.058; 6.310) were significantly associated with seropositivity to Chlamydophila spp. On the other hand, availability of veterinary service was identified as a protective factor (OR = 0.161, 95% CI (OR) = 0.051; 0.511).

Seroprevalence of Chlamydophila abortus infection in yaks (Bos grunniens) in Qinghai, China.

Chlamydophila abortus is an important amphixenosis which in a wide range of animals, associated with reproductive disorders in yaks. In order to assess the prevalence of this infection in yaks in Qinghai, China, a cross-sectional study was carried out, and a total of 674 serum samples were collected from June to October 2012 in six counties, and antibodies to C. abortus were examined by indirect hemagglutination (IHA) test. The overall seroprevalence of C. abortus in yaks was 17.66 % (119/674), and the seroprevalence of antibodies to C. abortus in yaks ranged from 11.82 to 28.43 % among the six different areas, and the difference was statistically significant (P < 0.05). The seropositivity of C. abortus infection in different age groups varied from 16.33 to 18.49 %, and prevalence in yaks of ≥3 year (18.49 %) was slightly higher than that in yaks of <3 year, but the differences among the age groups were not statistically significant (P > 0.05). The seroprevalence of C. abortus infection in male yak (16.8 %) was slightly lower than that in females (17.85 %), and the difference was not statistically significant (P > 0.05). So far, this is the first systematic and comprehensive investigation of C. abortus infectionin in yaks in this area.

Distinct intensity of host-pathogen interactions in Chlamydia psittaci- and Chlamydia abortus-infected chicken embryos.

Factors and mechanisms determining the differences in virulence and host specificity between the zoonotic agents Chlamydia psittaci and Chlamydia abortus are still largely unknown. In the present study, two strains were compared for their invasiveness, virulence, and capability of eliciting an immune response in chicken embryos. On breeding day 10, embryonated chicken eggs were inoculated with 5 × 10(4) inclusion-forming units. As shown by immunohistochemistry and quantitative real-time PCR, C. psittaci displayed a significantly better capability of disseminating in the chorioallantoic membrane (CAM) and internal organs than C. abortus. The higher infectious potential of C. psittaci in birds was underlined by significantly higher mRNA expression rates of essential chlamydial genes, such as incA, groEL (in CAM, liver, and spleen), cpaf, and ftsW (in CAM). Although the immune responses to both pathogens were similar, C. psittaci elicited higher macrophage numbers and a stronger expression of a subset of immune-related proteins. The data imply that invasiveness of Chlamydia spp. and propagation in the host are not solely dependent on the level of host immune response but, even to a greater extent, on the expression of bacterial factors related to virulence. The fact that C. psittaci has coped far better than C. abortus with the avian embryo's response by upregulating essential genes may be a key to understanding the mechanisms underlying host adaptation and etiopathology.

Prevalence and risk factors associated with Chlamydophila abortus infection in dairy herds in Jordan.

A cross-sectional study was carried out to determine seroprevalence and to identify risk factors associated with Chlamydophila abortus infection in 62 nonvaccinated dairy herds (671 cows) in Jordan between January and June 2007. Information regarding herd management was recorded through a personal interview with farmers. Antibodies against C. abortus were detected using an ELISA test kit. Chi-square analysis and multivariable logistic regression model were used to identify risk factors associated with C. abortus seropositivity. The true prevalence of antibodies against C. abortus in individual cows and cattle herds were 19.9 % and 66.3 %, respectively. Univariable Chi-square analysis revealed three variables with P ≤ 0.25 that were further offered to multivariable logistic regression analysis. Small-sized herds were identified as a risk factor for seropositivity to C. abortus, while sweeping followed by water hosing and using disinfectants were identified as protective factors. Cows in the age groups of >8 and ≤ 10 years old and >2 and ≤ 6 years old had the highest and lowest significant seroprevalence to C. abortus, respectively. Results of this study indicated that C. abortus is highly prevalent in Jordan's dairy herds and Chlamydophila infection could be controlled by applying strict biosecurity measures in the dairy farms.

Ovarian hydrobursitis in female camels (Camelus dromedarius): the role of Chlamydophila abortus and a trial for medical treatment.

The occurrence of Chlamydophila abortus in female camels affected with ovarian hydrobursitis and a trial for medical treatment were studied. A total of 111 cases were included in two experiments. In Experiment 1, sera from 51 affected cases were tested for C. abortus antibody using enzyme-linked immunosorbent assay (ELISA). In Experiment 2, 60 female camels affected with bilateral ovarian hydrobursitis were divided into treated and control groups (n = 30 each). Based on the bursal diameter, females of both groups were subdivided into those having small (< 5 cm), medium (5-7 cm) or large (> 7 cm) bursae. Treated group received 20 mg/kg body weight oxytetracycline intramuscular, 4% lotagen intrauterine, and 500 µg cloprostenol intramuscular. Controls did not receive any treatment. All females were observed for 90 days non-return rate (NRR) and calving rate (CR). Antibodies against C. abortus were observed in 44/51 (86.3%) of the affected females. The 90 days NRR of the treated and control groups were 13/30 (43.3%) and 0/30 (0.0%), respectively, (P = 0.001), while the CR were 10/30 (33.3%) and 0/30 (0.0%), respectively, (P = 0.01). Based on bursal size, the 90 days NRR were 11/15 (73.3%), 2/7 (28.6%) and 0/8 (0.0%) for treated females having small, medium and large bursa, while the CR were 9/15 (60%), 1/7 (14.3%), and 0/8 (0.0%), respectively, (P = 0.01). In conclusion, it seems that C. abortus may be responsible for the spreading of the ovarian hydrobursitis syndrome in dromedaries. Small sized bursa could be medically treated.

Protection of mice against Chlamydophila abortus infection with a bacteriophage-mediated DNA vaccine expressing the major outer membrane protein.

A bacteriophage-delivered DNA vaccine against Chlamydophila abortus was constructed by cloning a eukaryotic cassette containing the ompA gene (which expresses the Major Outer Membrane Protein) into a bacteriophage lambda vector. Four groups, each of 20 BALB/c mice were inoculated separately with the phage vaccine, a conventional DNA vaccine based on the same ompA expression cassette, a live attenuated vaccine (strain 1B) or the empty phage vector. The phage and DNA vaccines and empty phage vector were administered intramuscularly on days 0, 14 and 28; the attenuated vaccine was given once on day 0. Half the animals in each group were challenged on day 42 by intraperitoneal injection of live C. abortus and sacrificed on day 49. Phage-vaccinated mice developed moderate antibody levels against C. abortus and yielded higher levels of IFN-γ and IL-2 compared with the attenuated live vaccine group. Clearance of chlamydiae from spleens was significantly better in the attenuated vaccine group compared with the phage vaccine group, while both groups were significantly superior to the DNA vaccine and control groups (p<0.01). Although levels of protection in the mouse model were lower in phage-vaccinated animals, than in 1B vaccinated animals, phage vaccines offer several other advantages, such as easier handling and safety, potentially cheaper production and no chance of reversion to virulence. Although these are preliminary results in a model system, it is possible that with further optimisation immunization with phage vaccines may provide a novel way to improve protection against C. abortus infection and trials in large animals are currently being initiated.

Correlation of Chlamydia and Chlamydophila spp. IgG and IgM antibodies by microimmunofluorescence with antigen detection methods.

Correlation of serologic titers for Chlamydia trachomatis with other tests has been based on direct fluorescence antibody (DFA) testing and culture, but not on nucleic acid-based tests that are used for screening. We retrospectively reviewed the specificity of antibodies against C. trachomatis, Chlamydia psittaci, and Chlamydophila pneumoniae by microimmunofluorescence (MIF) when compared with DFA, culture, nucleic acid probe, and transcription-mediated amplification. Over a 6-year period, 226 cases had both MIF and one of these other methods performed for comparison. Agreement between C. trachomatis antigen or nucleic acid detection and MIF results was 87% (197/226). C. trachomatis serology had a negative predictive value of 98%, and 10.6% of cases were positive by serology and negative by antigen testing. Of the 13 patients who had a positive C. trachomatis antigen or nucleic acid test result, 9 had IgG and/or IgM titers highest against C. trachomatis, 3 had IgG titers highest against C. pneumoniae, and 1 had undetectable titers for the three chlamydial species. Twenty-five patients had positive IgG and/or IgM titers to C. trachomatis but negative antigen test results. Serologic testing can increase the sensitivity of detecting C. trachomatis infections.

Amniotic and allantoic fluids from experimentally infected sheep contain immunoglobulin specific for Chlamydophila abortus.

Chlamydophila abortus, the aetiological agent of enzootic abortion of ewes (EAE), replicates in trophoblast cells leading to their destruction and dissemination of the bacterium to foetal organs. To further understand the pathogenesis of EAE, amniotic and allantoic fluids were collected from experimentally infected pregnant ewes at 30 (7 samples from each fluid), 35 (8 samples from each fluid), 40 (10 samples from each fluid) and 43 (6 amniotic fluids and 7 allantoic fluids) days post-infection to determine pathogen numbers and other markers of infection. Whilst experimentally infected ewes had characteristic placental lesions, only two amniotic and seven allantoic fluid samples were positive for C. abortus by real-time PCR. In contrast, all amniotic and allantoic fluids were positive for immunoglobulin. Immunoglobulins were generally detected earlier in allantoic fluid than in amniotic fluid and the numbers of samples containing immunoglobulins increased as infection progressed. IgG in amniotic and allantoic fluids was shown to be specific for C. abortus, and reacted with the major outer membrane proteins, polymorphic outer membrane protein and macrophage infectivity potentiator protein. A comparison of two-dimensional immunoblots using purified IgG from the allantoic fluid, amniotic fluid, ewe serum and foetal serum of a C. abortus infected animal at 40 days post infection indicated a pattern of reactivity intermediate between that of the ewe serum and the foetal serum. Results suggest that a maternal source of immunoglobulin is predominant at 30 days post-infection but that foetal derived antibodies may be contributed at a later stage.

Co-administration of the polysaccharide of Lycium barbarum with DNA vaccine of Chlamydophila abortus augments protection.

Lycium barbarum polysaccharides (LBP) can stimulate moderate immune responses therefore could potentially be used as a substitute for oil adjuvants in veterinary vaccines. In the present study, it was shown that the isolated active component of LBP3a, combined with a DNA vaccine encoding the major outer membrane protein (MOMP) of Chlamydophila abortus, induced protection in mice against challenge. Sixty BALB/c mice were randomly assigned to 5 groups. Sub-fractions of polysaccharide LBP3a, at 12.5, 25 and 50 mg/kg concentrations, respectively, were mixed with a pCI-neo::MOMP (pMOMP) vaccine. Mice administrated with pCI-neo + LBP3a were served as a control. All mice were inoculated at day 0, 14, and 28, and challenged on day 44. The effects of LBp3a on serum antibody levels, in vitro lymphocyte proliferation, the activity of interleaukin-2 (IL-2), interferon-γ (IFN-γ), tumor necrosis factor α(TNF-α)and chlamydia clearance were determined. A combination of DNA vaccine and LBP3a induced significantly higher antibody levels in mice, higher T cell proliferation and higher levels of IFN-γ and IL-2. Mice immunized with DNA and LBP3a also showed significantly higher levels of chlamydia clearance in mice spleens and a greater Th1 immune response. The immunoenhancement induced by 25 mg/kg LBP3a is more effective than that induced by a 12.5 and 50 mg/kg. This implies that LBP3a at 25 mg/kg has a high potential to be used as an effective adjuvant with a DNA vaccine against swine Chlamydophila abortus.

Prevalence of antibodies against Chlamydophila abortus and Coxiella burnetii in goat herds in Poland.

An epidemiological study was carried out to determine the herd prevalence of Chlamydophila abortus and Coxiella burnetii antibodies in goats covered by a milk recording program in Poland. The survey took place in 2007 and 48 herds located in different parts of the country were involved. A representative sample from each herd was taken by a simple random sampling allowing to detect seropositivity of a herd on a 95% level of confidence. In total 918 goats were tested for specific antibodies against both germs with the use of enzyme-linked immunosorbent assays. In addition, history of reproductive failures was recorded in these herds. The survey revealed that the herd prevalence of C. abortus was 4.2% (2 herds) while no C. burnetii antibodies were found. Abortions were reported to be a problem in 80% of herds while repeating estrus was encountered in 46% of herds. Reproductive failure concerned two seropositive herds as well. Since the germ is present in the population, it has to be taken into consideration in diagnostic process. Nevertheless, the results of the present study indicate that C. abortus infection occurs infrequently in Polish goats. As no antibodies against C. burnetii were detected in the screened sample the risk of goat-to-human transmission of both bacteria in Poland seems to be very low.

Using CF0218-ELISA to distinguish Chlamydophila felis-infected cats from vaccinated and uninfected domestic cats.

Chlamydophila felis is a causative agent of acute and chronic conjunctivitis and pneumonia in cats. Cats can be vaccinated with killed or attenuated C. felis. However, current serodiagnostics cannot distinguish these cats from naturally infected cats. This causes difficulty of early diagnosis and seroepidemiological survey for C. felis. We previously reported that C. felis CF0218 can be used as a C. felis-infection-specific diagnostic antigen in experimentally infected and/or vaccinated cats. In this study, we evaluated an enzyme-linked immunosorbent assay using recombinant CF0218 as antigen (CF0218-ELISA) to detect anti-C. felis antibody in 714 sera of domestic cats whose histories of vaccination against C. felis are known. The 44 vaccinated cats were 93% negative using CF0218-ELISA; half of these scored positive by immunofluorescence assay (IFA) using C. felis-infected cells as antigen. The 670 non-vaccinated cats had CF0218-ELISA positivity rates that were statistically in agreement with IFA (18% vs. 21%). These results show that CF0218, which was identified as a C. felis-infection-specific antigen, is a useful serodiagnostic antigen to distinguish naturally C. felis-infected cats from vaccinated and non-infected cats.

Identification of immunologically relevant proteins of Chlamydophila abortus using sera from experimentally infected pregnant ewes.

Chlamydophila abortus is an intracellular pathogen and the etiological agent of enzootic abortion of ewes (EAE). C. abortus has a biphasic development cycle; extracellular infectious elementary bodies (EB) attach and penetrate host cells, where they give rise to intracellular, metabolically active reticulate bodies (RB). RB divide by binary fission and subsequently mature to EB, which, on rupture of infected cells, are released to infect new host cells. Pregnant ewes were challenged with 2 x 10(6) inclusion forming units (IFU) of C. abortus cultured in yolk sac (comprising both EB and RB). Serum samples were collected at 0, 7, 14, 21, 27, 30, 35, 40, and 43 days postinfection (dpi) and used to identify antigens of C. abortus expressed during disease. Additionally, sera from fetal lambs were collected at 30, 35, 40, and 43 dpi. All serum samples collected from experimentally infected pregnant ewes reacted specifically with several antigens of EB as determined by one-dimensional (1-D) and 2-D gel electrophoresis; reactive antigens identified by mass spectrometry included the major outer membrane protein (MOMP), polymorphic outer membrane protein (POMP), and macrophage infectivity potentiator (MIP) lipoprotein.

Efficacy of a new inactivated Chlamydophila felis vaccine in experimentally-infected cats.

A new inactivated and adjuvanted Chlamydophila felis vaccine was developed and its efficacy in cats was compared with that of commercially available inactivated and live vaccines. Two commercial vaccines conferred insufficient immunity on inoculated cats, as evaluated by antibody production and a challenge experiment, whereas cats administered the newly generated vaccine produced high-titre antibodies and acquired sufficient immunity. The cats immunised with the new vaccine revealed no or only mild clinical signs, and no chlamydiae were recovered from their tissue samples after exposure to a virulent C felis. However, they shed chlamydiae in their nasal and conjunctival secretions after challenge, as did those immunised with the commercial vaccines and the non-vaccinated controls. The newly developed vaccine caused no adverse reaction in the inoculated cats. These findings suggest that the new vaccine prepared here may be promising for practical use in controlling C felis infection in cats.

Polymorphic membrane proteins 1 and 7 from Chlamydophila felis are significant immunodominant proteins.

Chlamydophila felis is a common cause of conjunctivitis in cats. Greater understanding of C. felis infection and immunity and identification of protective antigens will facilitate improved vaccine design. Chlamydial polymorphic membrane proteins (Pmps) represent a family of homologous proteins of likely importance in chlamydial infection and immunity. To identify immunogenic C. felis Pmps, we generated recombinant C. felis Pmps (rPmps) and used these to detect serum antibody reactivity against Pmps arising during C. felis infection in cats. Sequencing of Pmp genes 1, 7, 13, 18, 19 and 20 from 3 laboratory strains of C. felis (K2487, 1497V and Cello) and alignment with the Fe/C-56 genome revealed high genetic identity in Pmp genes between strains. PCR products lacking the predicted N-terminal signal sequence peptide and C-terminal domain were generated, cloned and expressed in Escherichia coli prior to purification by nickel-agarose affinity chromatography. Serum samples from 4 cats collected up to 55 days post-inoculation with C. felis (K2487) were analysed by western blotting and rPmp-specific ELISAs for evidence of serum antibody reactivity against each rPmp. Strong serum antibody reactivity against rPmps 1 and 7, and weak heterogeneous serum immunoreactivity against rPmps 13, 19 and 20, were detected from 14 to 21 days post-infection (dpi), peaking at 28-35 dpi and tending to plateau thereafter. No significant serum antibody reactivity was detected against rPmp18. This study provides the first evidence that C. felis Pmps 1 and 7 are likely to represent immunodominant proteins and recommends investigation of their potential as serodiagnostic antigens and novel vaccine candidates.

Serological analysis of Chlamydophila abortus in Australian sheep and implications for the rejection of breeder sheep for export.

OBJECTIVE: To examine the incidence of positive results in a complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) for Chlamydophila abortus in Australian sheep and how this incidence differs with state of origin, age, sex, breed and property. To examine the consequences in relation to rejection of breeder sheep for export. DESIGN: Collection of blood samples from 891 sheep on 109 properties in southern Australia. All samples had a unique, coded property identification. PROCEDURE: The samples were tested using the Institut Pourquier Chlamydophila abortus antibody ELISA (rELISA) and a CFT. Residual maximum likelihood analyses of the sample to positive ratio of the corrected optical density for the rELISA and generalised linear mixed model analyses of the CFT outcomes were carried out. RESULTS: The sample to positive ratio of the corrected optical density values of the rELISA did not differ between sex, age, breed or state of origin, but differed greatly between properties. The CFT outcome did not differ between age, breed or state of origin, but differed greatly between properties and was more often positive with rams than with ewes. CONCLUSION: Positive outcomes to C. abortus antibody tests are very common in Australia. Rams have a particularly high incidence of positive results with the CFT. Rejection of sheep and property consignments is likely to be very common with all tests and situations examined except for the CFT (at 1:32 dilution) in ewes.

Investigation of Chlamydiaceae in semen and cauda epididymidis and seroprevalence of Chlamydophila abortus in breeding bulls.

BACKGROUND: Reproductive disorders associated with chlamydial infection have been reported worldwide in cattle and there are indications of potential venereal transmission. METHODS: Semen samples from 21 dairy bulls and cauda epididymidis tissue samples from 43 beef bulls were analysed for chlamydial agent by real-time polymerase chain reaction (PCR) including an internal amplification control (mimic). Additionally, presence of antibodies against Chlamydophila (Cp.) abortus among the bulls was investigated with the commercial Pourquier ELISA Cp. abortus serum verification kit. RESULTS: No chlamydial agent was detected by PCR in either the semen samples or in the tissue samples. Additionally, no antibodies against Cp. abortus were detected. CONCLUSIONS: The results suggest that Cp. abortus is very rare, or absent in Swedish bulls and thus the risk for venereal transmission of chlamydial infection through their semen is low. However, because Chlamydophila spp. infection rates seem to differ throughout the world, it is essential to clarify the relative importance of transmission of the infection through semen on cattle fertility.

Seroprevalence survey of Chlamydophila abortus infection in breeding goats on commercial farms in the Otavi Veterinary District, northern Namibia.

A total of 1 076 sera from breeding goats were randomly collected from 24 different farms and tested with CHEKIT®-ELISA (IDEXX Laboratories B.V., 1 119 NE Schiphol-Rijk, Nederland) for antibodies against Chlamydophila abortus. The farms were divided into two categories of twelve farms each,based on their previous history of observed abortions over the previous 12 months: those with low (< 5%) levels of abortion and those with high (≥ 5%) levels of abortion. The farmers were also interviewed on their level of awareness about chlamydophilosis, its zoonotic importance and vaccination measures against the disease. The study detected overall seroprevalence levels of 25% for the farms and 8% for the individual animals (at 95% confidence). A total of six out of twentyfour farms (25%) had at least one positive breeding animal. Only five out of the twenty-four (20.8%)farmers interviewed were aware of chlamydophilosis and its zoonotic dangers. None of the 24 farmers interviewed practised any vaccination against chlamydophilosis. There was a significantly higher number of seropositive animals from farms with high levels of abortion, compared to those animals from farms with low levels of abortion (p = 0.0001). This study underscores the need for a higher level of farmer awareness and training on chlamydophilosis and its zoonotic dangers.