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PubMed1994

Monoclonal antibody to a major outer membrane protein of feline Chlamydia psittaci: antibody specificity and anti-idiotype antibody production.

A murine monoclonal antibody, F1-8, was developed against the purified major outer membrane protein (MOMP) of Chlamydia psittaci feline pneumonitis (FPn). F1-8 showed a serotype-specific activity against intact Fpn elementary bodies in a micro-immunofluorescence assay. In immunoblot, F1-8 reacted only with the Fpn MOMP but did not react with the MOMPs from other strains of C. psittaci and C. trachomatis. F1-8 neutralized Fpn infectivity in L929 cell culture in a dose-dependent and complement independent fashion. These results suggested that the monoclonal antibody (mAb) binds with an epitope on the MOMP region that is exposed at the cell surface and plays an important role in FPn infection. Polyclonal anti-idiotype (anti-Id) antibodies to mAb F1-8 were elicited by F1-8 coupled to keyhole limpet hemocyanin. These anti-Id antibodies inhibited F1-8 binding to FPn MOMP.

Animals↗
PubMed1995

Chlamydia psittaci infections: a review with emphasis on avian chlamydiosis.

In the first part of this article the general characteristics of Chlamydia psittaci namely the history, taxonomy, morphology, reproductive cycle, metabolism and genetics are reviewed. For the taxonomy in particular, a considerable amount of new information has become available in recent years, following the application of monoclonal antibodies and restriction enzymes. Using these techniques isolates of Chlamydia psittaci from birds have been subdivided in different serovars, a number of isolates have been classified in a new species (Chlamydia pecorum) and isolates from animals have been classified as Chlamydia trachomatis. In the second part of the article, the current knowledge on avian chlamydiosis is summarized. Emphasis is put on clinical signs, lesions, pathogenesis, epizootiology, immunity, diagnosis, prevention and treatment. Also the public health considerations are reviewed. It is concluded that the diagnosis of avian chlamydiosis is laborious and that there is still a need for more accurate, simple and rapid diagnostic tools, both for antigen and antibody detection in various species of birds.

Animals↗
PubMed1995

Chlamydia psittaci in turkeys: pathogenesis of infections in avian serovars A, B and D.

At 7 days of age, 4 groups, each of twenty specific pathogen free turkeys kept in isolation units were inoculated by aerosol with the Texas Turkey strain (avian Chlamydia psittaci serovar D), strain 92/1293 (avian Chlamydia psittaci serovar D), strain 84/55 (avian Chlamydia psittaci serovar A) or strain 89/1326 (avian Chlamydia psittaci serovar B). A fifth group of 4 specific pathogen free turkeys were sham inoculated controls. At daily intervals for 10 days and then twice weekly up to 34 days post infection, one bird in each group was killed and the target tissues and cells for replication and the sequence of events of serovar A, B and D infections was examined. In these turkeys, the primary site of replication was the respiratory tract. Chlamydial replication could be detected in the respiratory tract on day 1 post inoculation (p.i.) for group A, on day 3 p.i. for group B and on day 1 to 2 p.i. for groups D1 and D2. Subsequently, there was chlamydaemia and localisation in the digestive tract, in one or more parenchymatous organs, in the pericardium and in the conjunctivae. Specific immunoperoxidase staining revealed chamydiae in these organs in epithelial cells and in monomorphonuclear cells in all infected groups. The monomorphonuclear cells were identified as macrophages by double immunofluorescence staining. Chlamydiae were present in the same tissues for serovars A and D, but could not be demonstrated in proventriculus, duodenum, pancreas, ovaries and testes for serovar B. Furthermore, the intensity of replication was similar for all serovars. However, for serovar B in comparison with the other serovars, the bacteria appeared in most tissues 1 to 6 days later and the maximal replication in these tissues occurred 3 to 4 days later.

Aerosols↗
PubMed1996

Evaluation of Chlamydia psittaci subfraction and subunit preparations for their protective capacities.

A mouse toxicity potency test was used to evaluate the protective capacity of Chlamydia psittaci (Baker strain) subfraction and subunit preparations. The elementary body (EB), Chlamydial outer membrane complex (COMC), major outer membrane protein (MOMP), lipopolysaccharide (LPS), and MOMP mixed together with LPS were utilized as antigens in vaccine preparation. The EBs and COMCs were prepared by centrifugation and detergent extraction as previously described for Chlamydia trachomatis. Isolation and purification of MOMP and LPS were achieved by subjecting infected tissue culture fluids to polyacrylamide gel electrophoresis and subsequent elution from the gel. Chromatographic purification of the MOMP from EBs resulted in a preparation which also contained minor quantities of LPS. After vaccination and intraperitoneal challenge with C. psittaci (Cello strain), the MOMP presented in EBs, COMCs or in a purified form, was capable of protecting mice from death, whereas LPS did not demonstrate this capability. Further comparison of the protective capacity of these antigens indicated that the electrophoretically purified MOMP was the least protective when suboptimal levels of antigens were administered to mice. However, the level of protection afforded by electrophoretically purified MOMP could be increased to that observed with the chromatographically purified MOMP by the addition of electrophoretically purified LPS to the vaccine. These results suggest that the MOMP is the major protective antigen of C. psittaci and that the LPS component, although not protective on its own, may play a role in the potentiation of a protective immune response.

Animals↗
PubMed1996

Diversity among abortion strains of Chlamydia psittaci demonstrated by inclusion morphology, polypeptide profiles and monoclonal antibodies.

Twenty eight C. psittaci abortion strains had been previously classified in to 4 immunologically distinct groups on the basis of cross-protection experiments in a mouse model. To identify the molecular basis of their immunological divergence 4 representative strains were investigated by cellular, molecular and immunological techniques. An identical pattern was obtained by Alul digestion of the amplified major outer membrane protein gene (MOMP) by the polymerase chain reaction (PCR) of the 4 strains. However, inclusion morphology and polypeptide profiles clearly distinguished one strain, named LLG, and its homologous strain POS from the other prototypes by the presence of a unique protein at 26.5 kDa and the absence of a polypeptide at 23 kDa. Six out of 10 monoclonal antibodies (mAbs) raised against abortion strains failed to react with inclusions of the 2 strains. All 6 mAbs reacted with the chlamydial outer membrane complex (COMC). Two of these mAbs, one against the MOMP and one against an antigen at 90 kDa, did not react with immunoblots of LLG and POS. The data provide direct demonstration of the existence of strain variation in the field and classify strains LLG and POS as a distinct C. psittaci serotype 1-subtype. The antigenic diversity among abortion strains should be taken into consideration when designing a subunit vaccine.

Abortion, Veterinary↗
PubMed2005

Is there an association between ocular adnexal lymphoma and infection with Chlamydia psittaci? The University of Rochester experience.

Various subsets of extranodal marginal zone lymphomas of mucosa-associated lymphoid tissues (MALT lymphomas) have been associated with infectious organisms. Most notable of these is the association of gastric MALT lymphomas with Helicobacter pylori infection. In a recent publication Ferreri et al. [Ferreri AJ, Guidoboni M, Ponzoni M, De Conciliis C, Dell'Oro S, Fleischhauer K, et al. Evidence for an association between Chlamydia psittaci and ocular adnexal lymphomas. J Natl Cancer Inst 2004;96:586-94] reported the presence of C. psittaci DNA in 80% of 40 ocular adnexal lymphomas. Similar to the gastric MALT lymphoma data, a subset of these patients responded well to antibiotic treatment. We analyzed a set of ocular adnexal lymphomas and benign (non-neoplastic) lesions for evidence of C. psittaci DNA in patients from New York State. No evidence of C. psittaci DNA was seen in seven MALT-type ocular adnexal lymphomas, four non-MALT ocular lymphomas, one Langerhans histiocytosis, and five reactive lymphoproliferations. We eliminated several possible reasons that would cause our study to fail to find C. psittaci DNA, including the presence of PCR inhibitors, inadequate template DNA, and sequence diversity in the target region in C. psittaci. The positive data were based primarily on patients from Italy, while our study involved only patients living in the Northeastern United States. This would suggest possible geographic differences in the etiology of ocular adnexal lymphomas.

Adolescent↗
PubMed2004

Seroprevalences for ovine enzootic abortion in Switzerland.

Our aim was to assess the seroprevalence of Chlamydophila (Cd) abortus (Chlamydia psittaci serotype 1), denoted ovine enzootic abortion (OEA), in the Swiss sheep population. A competitive enzyme-linked immunosorbent assay (cELISA) was adapted for the investigation of pooled serum samples (pool approach) and receiver-operator characteristic (ROC) analysis was applied to define the cut-off of the pool approach. At a cut-off value of 30% inhibition, the flock-level pooled sensitivity and specificity were 92.9% and 97.6% when compared to classifying the flock based on individual-animal samples. Subsequently, sera from 775 randomly selected flocks out of 11 cantons of Switzerland were investigated using the pool approach. The cantons included in the study represented 72% of the Swiss sheep flocks and 76% of Swiss sheep population. Antibodies against Cd. abortus were found in almost 19% (144) of the 775 examined sheep flocks. Test prevalences were adjusted for the imperfect test characteristics using the Rogan-Gladen estimator and Bayesian inference. Seroprevalence was highest (43%) in the canton Graubunden. In the remaining 10 cantons the seroprevalence ranged from 2 to 29%. The cELISA in combination with testing pooled sera and statistical methods for true prevalence estimation provided a good survey tool at lower costs and time when compared to other approaches.

Abortion, Veterinary↗
PubMed2004

Influence of maternal antibodies on Chlamydophila psittaci-specific immune responses in turkeys elicited by naked DNA.

Plasmid DNA (pcDNA1::MOMP D) expressing the major outer membrane protein (MOMP) of an avian Chlamydophila psittaci serovar D strain was tested for its ability to induce protective immunity against C. psittaci challenge in the presence of maternal antibodies. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was used. Following pcDNA1::MOMP vaccination, both T helper and B cell memory were primed. However, high maternal antibodies titres affected the induction of vaccine-specific antibody responses as assessed by MOMP-specific antibody levels in enzyme-linked immunosorbent assay (ELISA). Cell-mediated immunity was unaltered as demonstrated by the significantly heightened proliferative responses of peripheral blood lymphocytes (PBL) following vaccination. DNA vaccination could significantly reduce clinical symptoms, pharyngeal and cloacal excretion as well as Chlamydophila replication, even in the presence of maternal antibodies.

Administration, Intranasal↗
PubMed2005

Protection of turkeys against Chlamydophila psittaci challenge by DNA and rMOMP vaccination and evaluation of the immunomodulating effect of 1 alpha,25-dihydroxyvitamin D(3).

Plasmid DNA expressing the major outer membrane protein (MOMP) of an avian Chlamydophila psittaci serovar D strain and recombinant MOMP (rMOMP) with or without the immunomodulating adjuvant 1 alpha,25-dihydroxyvitamin D(3) have been tested for their ability to elicit an immune response and induce protection in turkeys against challenge with the same serovar. Three vaccination strategies were compared: priming and boosting with either pcDNA1::MOMP or rMOMP and priming with pcDNA1::MOMP followed by rMOMP boosting. Turkeys primed with pcDNA1::MOMP showed significant protection against Cp. psittaci challenge, turkeys primed with rMOMP did not. The steroid hormone 1 alpha,25-dihydroxyvitamin D(3) augmented serum and mucosal antibody titres. However, higher antibody titres were not related to better protection and even had a negative effect on especially bacterial excretion.

Adjuvants, Immunologic↗
PubMed2005

Key role of Chlamydophila psittaci on Belgian turkey farms in association with other respiratory pathogens.

Two hundred turkey sera from eight Belgian and two French farms were tested for the presence of antibodies against avian pneumovirus (APV), Ornithobacterium rhinotracheale (ORT), Mycoplasma gallisepticum, Mycoplasma meleagridis and Chlamydophila psittaci. At slaughter, C. psittaci, APV and ORT antibodies were detected in 94, 34 and 6.5% of the turkeys, respectively. No antibodies against M. gallisepticum or M. meleagridis were present. Additionally, turkeys on three Belgian farms were examined from production onset until slaughter using both serology and antigen or gene detection. All farms experienced two C. psittaci infection waves, at 3-6 and 8-12 weeks of age. Each first infection wave was closely followed by an ORT infection starting at the age of 6-8 weeks, which was still detectable when the second C. psittaci infection waves started. Animals on farm A were not vaccinated against APV leading to an APV subtype B outbreak accompanying the first C. psittaci infection wave. Despite subtype A APV vaccination on farms B and C, the second C. psittaci infection waves were accompanied (farm B) or followed (farm C) by a subtype B APV infection. On all farms respiratory signs always appeared together with a proven C. psittaci, APV and/or ORT infection. This study suggests an association between C. psittaci, APV and ORT, and indicates the multi-factorial aetiology of respiratory infections in commercial turkeys. All three pathogens should be considered when developing prevention strategies for respiratory disease.

Age Factors↗
PubMed2005

Pathogenic interactions between Chlamydophila psittaci and avian pneumovirus infections in turkeys.

Both Chlamydophila psittaci and avian pneumovirus (APV) are highly prevalent in Belgian turkeys and might contribute to the respiratory disease complex observed in turkeys. Initial outbreaks of chlamydiosis occur mostly at the age of 4-8 weeks, often accompanied by an APV infection in APV non-vaccinated farms. Regardless APV vaccination, breakthroughs of APV infection from 8 weeks on do occur, a period when also a second C. psittaci infection appears. Therefore, this study examined the pathogenicity of an APV superinfection in C. psittaci predisposed turkeys. Turkeys were infected with C. psittaci, APV or with C. psittaci followed by APV. Simulating the impact of an APV infection during the acute phase or latent phase of a C. psittaci infection, turkeys have been infected with APV at 1 and 5 weeks post C. psittaci infection, respectively. APV infection during the acute phase of a C. psittaci infection aggravates the severity of clinical signs, macroscopic lesions, pharyngeal APV excretion and histological tracheae lesions. In contrast, no clear interaction could be established after APV infection in latently C. psittaci infected specific pathogen-free (SPF) turkeys. This study clearly demonstrates the exacerbating role of APV during acute C. psittaci infection, which can play an important role in the respiratory disease complex of turkeys.

Acute Disease↗
PubMed2006

Chlamydophila psittaci in free-living Blue-fronted Amazon parrots (Amazona aestiva) and Hyacinth macaws (Anodorhynchus hyacinthinus) in the Pantanal of Mato Grosso do Sul, Brazil.

Chlamydophila psittaci (C. psittaci) infection was evaluated in 77 free-living nestlings of Blue-fronted Amazon parrots (Amazona aestiva) and Hyacinth macaws (Anodorhynchus hyacinthinus) in the Pantanal of Mato Grosso do Sul, Brazil. Tracheal and cloacal swab samples from 32 wild parrot and 45 macaw nestlings were submitted to semi-nested PCR, while serum samples were submitted to complement fixation test (CFT). Although all 32 Amazon parrot serum samples were negative by CFT, cloacal swabs from two birds were positive for Chlamydophila DNA by semi-nested PCR (6.3%); these positive birds were 32 and 45 days old. In macaws, tracheal and cloacal swabs were positive in 8.9% and 26.7% of the samples, respectively. Complement-fixing antibodies were detected in 4.8% of the macaw nestlings; macaw nestlings with positive findings were between 33 and 88 days old. These results indicate widespread dissemination of this pathogen in the two evaluated psittacine populations. No birds had clinical signs suggestive of chlamydiosis. To the best of our knowledge, this is the first report on C. psittaci in free-living Blue-fronted Amazon parrots and Hyacinth macaws in Brazil.

Amazona↗
PubMed2006

A pmp genes-based PCR as a valuable tool for the diagnosis of avian chlamydiosis.

In a previous study we described the use of a new set of PCR primers (CpsiA/CpsiB) specific of the conserved pmp-family genes of Chlamydophila abortus as an efficient tool for the detection of these bacteria in ruminants including also preliminary results on avian strains. In this work, the use of this set of primers was extended to representative strains of the six major avian serovars (serovars A-F) and to field isolates of C. psittaci. For all the studied representative strains, using purified genomic DNA as a template, CpsiA/CpsiB primers allowed, as observed for C. abortus, a minimal 10-fold PCR signal increase compared to the one observed with ompA specific primers. In comparison to primers targeting the 16S-23S rRNA intergenic spacer, similar or increased sensitivity was observed depending on the strain. All the field isolates were amplified with CpsiA/CpsiB primers. On clinical samples, our primers are the best among those tested for detection of C. psittaci by simple conventional PCR. RFLP experiments performed using PCR fragments amplified with the CpsiA/CpsiB primers gave promising results demonstrating that these primers may provide an interesting tool for molecular typing when the bacterium cannot be grown from pathological samples.

Animals↗
PubMedDate not supplied

The role of Branhamella ovis, Mycoplasma conjunctivae and Chlamydia psittaci in conjunctivitis of sheep.

Smooth (S) and rough (R) types of Branhamella ovis were used alone and in conjunction with Mycoplasma conjunctivae and Chlamydia psittaci in attempts to produce conjunctivitis in lambs. This was only successful with one S-type previously isolated from a clinical case of conjunctivitis. In experimental mixed infection, B. ovis contributed to the severity of ovine keratoconjunctivitis attributable to infection with Mycoplasma conjunctivae. When B. ovis and C. psittaci were inoculated into the conjunctival sac mild conjunctivitis was produced but C. psittaci could not be re-isolated.

Animals↗
PubMedDate not supplied

Clinical and immunological responses of ewes following vaccination with an experimental formalin-inactivated Chlamydia psittaci (ovis) vaccine and subsequent challenge with the live organism during pregnancy.

The protection afforded by an experimental, killed, adjuvanted vaccine derived from the A22 strain of Chlamydia psittaci (ovis) against ovine enzootic abortion was studied. The vaccine was used undiluted (group A), at a dilution of 10(-3) (group B) and at a dilution of 10(-6) (group C). A fourth control group (group D) was inoculated with all components of the vaccine except the chlamydial antigen. A group of rams (group R) was also vaccinated with the chlamydial antigen diluted to 10(-3). Animals were challenged 70 days after mating with the A22 strain of C. psittaci (ovis) and were studied throughout pregnancy and the subsequent lambing period. Their cell-mediated immune responses were examined using a skin test and their humoral immune responses were studied using an ELISA. Tests for excretion of chlamydiae in their faeces and genital tract during pregnancy and after parturition and in the faeces of their lambs were made. The reproductive performance of the ewes was assessed by calculating the average weight of lambs produced per ewe in each group. The experimental vaccine protected the ewes in groups A and B against challenge with C. psittaci (ovis) as none showed clinical signs of OEA or excreted chlamydiae. The average weight of lambs produced per ewe in both groups was greater than 4 kg. Both groups seroconverted after vaccination but not all of them were positive to the skin test. The experimental vaccine at 10(-6) dilution of antigen did not protect the ewes as three of 10 ewes displayed clinical OEA and excreted chlamydiae in the products of parturition.(ABSTRACT TRUNCATED AT 250 WORDS)

Abortion, Veterinary↗
PubMedDate not supplied

Protection of ewes vaccinated with A22 strain Chlamydia psittaci (ovis) against challenge in pregnancy with homologous and heterologous strains of the organism.

Fifty ewes were randomly divided into four groups. Groups A and B were vaccinated with an experimental vaccine derived from the A22 isolate of Chlamydia psittaci (ovis), an isolate known to cause ovine enzootic abortion (OEA). Groups C and D were unvaccinated controls. In mid-pregnancy, animals in group A and C were challenged with live A22 C. psittaci (ovis) and those in B and D were challenged with a field isolate of the organism (BS) against which the commercially available A22 vaccine appeared to offer poor protection. In group A, three animals showed clinical signs of OEA and six excreted chlamydiae. In group B, five ewes showed clinical signs of OEA and excreted chlamydiae. In group C, three ewes had clinical signs of OEA but seven excreted chlamydiae. In group D, all 11 ewes showed clinical signs of OEA and excreted chlamydiae in the products of parturition. This group produced only four live lambs with an average weight of viable lamb per ewe of 1.4 kg, whereas the other groups each produced 12 or 13 lambs with an average weight of viable lamb per ewe of more than 4 kg. The BS isolate was much more virulent than the A22 isolate for unvaccinated, pregnant ewes. However, the A22 vaccine offered significant protection against the heterologous BS isolate although on this occasion it did not appear to alter the course of disease produced by the less pathogenic A22 isolate.

Abortion, Veterinary↗
PubMedDate not supplied

Use of the IDEIA ELISA to detect Chlamydia psittaci (ovis) in material from aborted fetal membranes and milk from ewes affected by ovine enzootic abortion.

The IDEIA ELISA was used to detect Chlamydia psittaci (ovis) antigen in ewes' milk to which were added serial dilutions of chlamydiae titrated as inclusion forming units (ifus) in McCoy cell tissue culture. The test was able to detect as few as 35 ifus/ml of the organism. The ELISA was then used to detect chlamydial antigen in fetal membranes and milk from ewes clinically affected with ovine enzootic abortion (OEA). The results were compared with results of isolation of chlamydiae in McCoy cell tissue culture from the same material. The fetal membranes of 17 of 19 ewes were positive for chlamydia when tested with the ELISA but chlamydia could be cultured from only 15 of them. Milk samples from 26 ewes which had aborted between 1 and 34 days previously were tested: chlamydiae could not be cultured from any of them and only one was positive when tested by the ELISA. The results show that the IDEIA ELISA is a sensitive test for the detection of C. psittaci (ovis) antigens. The positive results to this test for the three samples from which chlamydiae could not be cultured suggest that the test is not as specific as culture or that it detected dead organisms. Chlamydiae do not appear to be excreted in the milk of ewes affected with OEA.

Abortion, Veterinary↗