Epidemiological investigation of Chlamydophila psittaci in pigeons and free-living birds in Croatia

During 2003, 278 adult pigeons (Columba livia) and 54 birds of 11 other free-living species were caught in the various locations in the City of Zagreb, Croatia. Sera from 182 pigeons were tested for the presence of antibodies against Chlamydophila (C.) psittaci by ELISA test and 174 of them (95.6%) were found positive. Because of the high positivity rate in sera, cloacal swabs of 278 pigeons as well as 54 other species of free-living birds were tested for the presence of C. psittaci antigen. Fourty-four of the 278 pigeons (15.83%) were antigen positive, whereas all 54 of the wild birds were negative. Antigen-positive pigeons were euthanised and examined pathomorphologically and cytologically. Findings of specific antibodies and antigen of C. psittaci confirmed the high rate of infection among urban pigeons in the City of Zagreb, fortunately not among other free-living birds. Although the pigeon serovars of C. psittaci are considered to be of moderate pathogenicity for humans, the identification of 15.8% antigen-positive birds represents a potential source of infection to humans, especially for elderly people and immunodeficient patients, as well as for poultry in the Zagreb city area.     

Evidence of Chlamydophila psittaci infection in captive Amazon parrots in Brazil.

The prevalence of Chlamydophila psittaci (formerly Chlamydia psittaci) infection was assessed in 95 apparently healthy, captive Amazon parrots from three breeder collections in southeastern and west-central Brazil. Cloacal swabs from 95 birds were tested for chlamydial antigen, which was detected by direct immunofluorescence (DIF), and serum samples from 44 of these birds were tested for antibodies to C. psittaci using an enzyme-linked immunosorbent assay. The prevalences of active infection as detected by DIF were 16.7%, 22.2%, and 56.1%, and seroprevalences were 100%, 87.5%, and 60% in flocks A, B, and C, respectively. We can therefore infer that C. psittaci may be widespread in captive parrot populations in Brazil.     

Chlamydial infections in feral pigeons in Europe: Review of data and focus on public health implications

Feral pigeons (Columba livia domestica), which thrive in most European towns and cities, are commonly infected with the zoonotic bacterium Chlamydophila psittaci, the agent of psittacosis (also known as ornithosis) in humans. A number of surveys carried out over the last thirty years across Europe have detected high seropositivity values and high percentages of infection in feral pigeon populations. Overall, when considering data from 11 European countries, seropositivity values to C. psittaci in the sampled populations ranged from 19.4% to 95.6%. In most surveys, the complement fixation test was used, and antibodies were detected in 19.4-66.3% of the samples, with a median of 46.1%. Indirect immunofluorescence and ELISA tests were employed less frequently, but led to the detection of higher percentages of seropositivity (23.7-67.7% and 35.9-95.6%, respectively). Attempts to grow C. psittaci in cell culture or embryonated chicken eggs were successful in 2-42.3% and 0-57.1% of samples, respectively, antigen detection methods were positive in 2.3-40% of samples, while conventional PCR and real-time PCR using different genomic targets detected the organism in 3.4-50% of samples. Twenty-five C. psittaci isolates from pigeons were typed as ompA genotype B (n=14), E (n=10) and E/B (n=1). The huge increase of feral pigeon populations in Europe is a major cause of concern for the detrimental effect of pigeon droppings on environmental hygiene, in addition to the extensive damage due to the fouling of buildings and monuments. The most important pathogenic organism transmissible from feral pigeons to humans is C. psittaci, with 101 cases of disease reported in the literature. Exposure to C. psittaci-contaminated dust, direct contact with pigeons through handling and, to a lesser extent, through pigeon feeding have been identified as hazardous exposures in more than half of the human cases, while loose or transient contacts with feral pigeons have been mentioned in about 40% of the cases. Education initiatives as to the communication of a health risk resulting from contact with pigeons and pigeon excreta should primarily be targeted at individuals who may be exposed to C. psittaci-contaminated dust, such as demolition/construction workers. Recommendations to this category of workers include wearing protective clothes with hoods, boots, gloves and air filter face masks when removing pigeon faeces from roofs, garrets and buildings, especially if working indoors. Monitoring for C. psittaci infections in these workers over time should also be considered. Children should be warned not to handle sick or dead pigeons, and immunocompromised individuals should be advised to carefully limit their contact to feral pigeons. Culling of pigeons by shooting or poisoning is both unethical and ineffective as the place of the killed birds in the population is quickly filled by new juveniles or immigrating birds from neighbouring areas. Pigeon-deterring systems, such as nets and plastic or metal spikes applied to buildings and monuments will prevent their fouling, and the administration of contraceptive drugs may allow size regulation of the pigeon populations. Nevertheless, the measure that will ultimately lead to permanent reduction and will establish healthy sustainable populations is the restriction of indiscriminate feeding by pigeon lovers. The erection of dovecotes and artificial breeding facilities should be considered for providing shelter and a balanced diet to the birds, as well as a chance of interaction for pigeon lovers in a hygienically controlled environment.     

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.     

Long-term study of Chlamydophilosis in Slovenia

Immune reactivity for Chlamydophila (C.) psittaci in Slovenia was monitored in parrots, canaries, finches and nine species of recently captured free-living birds (house sparrows, Eurasian goldfinches, tree sparrows, chaffinches, European greenfinches, European serines, Eurasian siskins, Eurasian linnets and Eurasian bullfinches) in the period from 1991 to 2001. In subsequent years, specific IgG antibodies were found using immunofluorescence in parrots (0.7-53.6%), canaries (0.0-3.5%), finches (0.0-5.7%) and in captured free-living birds (33.3% of Eurasian goldfinches in 1994). An experimental infection with C psittaci was performed in order to study clinical signs and pathological changes in canaries and finches. The C. psittaci strain used for experimental infection was isolated from a cockatiel (Nymphicus hollandicus). Chlamydial DNA was extracted from clinical material followed by RFLP-PCR analysis. Infection of canaries and finches was confirmed in organ smears by direct immunofluorescence and a modified Gimenez staining method. In addition, serological tests of indirect immunofluorescence and complement fixation were applied. However, in spite of positive immunological reaction there were no clinical signs three weeks after infection. The present study includes results of a serological survey of persons belonging to the most important risk groups (breeders, pet shopkeepers and veterinarians). The results of microimmunofluorescence to identify the presence of specific antibodies and correlation between appearance of infection in birds and important risk groups are presented. Out of 143 persons belonging to the high-risk group we found 10 (7%) persons who were immunologically positive. Testing of two successive samples was used to demonstrate an increase in IgG and IgA titres in human sera. However, IgM, which is indicative of acute infection, could not be detected.     

More than classical Chlamydia psittaci in urban pigeons

In the literature, studies of Chlamydia infection in birds have usually been confined to the search for Chlamydia (C., formerly Chlamydophila) psittaci, so that little is known about the presence of other chlamydial agents. In the present study, cloacal swabs and faeces samples of urban pigeons have been examined by real-time PCR, DNA microarray assays and partial ompA sequencing. Whilst C. psittaci was the predominant chlamydial agent in this pigeon population (75.8% of all Chlamydiaceae positives), the combined use of highly specific and sensitive molecular assays facilitated the detection of atypical serovars of C. psittaci, as well as other species of Chlamydia, such as C. abortus. Detection of C. pecorum and C. trachomatis from an avian host is reported here for the first time. Rather unexpectedly, 19.5% of all Chlamydiaceae-positive cases turned out to be infected with non-classified organisms. The considerable prevalence of these novel agents raises the question of their epidemiological importance and possible role as pathogens. Future surveys in domestic and wild birds will have to take the extended variety of chlamydial organisms into account.     

Serological response to pgp3 protein in animal and human chlamydial infections

Specific antibodies to plasmid-encoded protein pgp3 are known to be encountered in human Chlamydia (C.) trachomatis infections. In order to verify whether antibodies to this protein could be developed in animals infected with plasmid-carrying chlamydial strains, 454 animal sera were examined using a home-made pgp3 protein ELISA and Western blots (WB) of recombinant pgp3 protein from Chlamydophila (Cp.) psittaci. Likewise, 50 human sera were tested by ELISA and WB of recombinant pgp3 from C. trachomatis. The reactivity against pgp3 protein was compared to the reactivity against chlamydial elementary bodies (EBs) detected by microimmunofluorescence (MIF) test. The presence of pgp3-specific antibodies was demonstrated in most ducks and pigeons with Cp. psittaci infection detected by MIF, as well as in the majority of symptomatic cats and pigs infected with Cp. felis and C. suis, respectively, which reacted at high titres to Cp. felis and C. suis EBs by MIF. Moreover, most of the sera collected from patients with C. trachomatis culture-confirmed infection and seropositive to C. trachomatis by MIF, presented antibodies specific to C. trachomatis pgp3 recombinant protein. Therefore, pgp3 protein could be a useful marker of chlamydial infections in animals, as well as in humans.     

Health status of free-living pigeons (Columba livia domestica) in the city of Ljubljana

In the year 2000 an epidemiological research was undertaken on the health status of free-living pigeons in the city of Ljubljana, Slovenia. A total of 139 pigeons were captured and examined for the most common bacterial, viral, and parasitic diseases. Serum samples, oropharyngeal and cloacal swabs as well as samples of droppings and feathers were taken from the captured birds. Antibodies to paramyxovirus type 1 were found in 84.2% of the sera examined, and 23.7% of birds were serologically positive to Chlamydophila psittaci. Antibodies to avian influenza virus were not detected. Salmonella spp. were isolated from 5.7% of the cloacal swabs. Trichomonas gallinae was clinically suspected and then microscopically confirmed using oropharyngeal swabs in 7.9% of examined birds. Eimeria spp. was identified in 71.9%, Capillaria sp. in 26.6% and Ascaridia columbae in 4.3% of droppings samples examined. Of the ectoparasites, Columbicola columbae and Campanulotes bidentatus compar were found.     

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.     

Epidemiology of Chlamydia psittaci Infection in Racing Pigeons and Pigeon Fanciers in Beijing,China

Over 3 million racing pigeons (Columba livia) are registered in Beijing City Center for gambling purposes. During 2008–2010, we evaluated the occurrence and prevalence of Chlamydia psittaci in racing pigeons as well as the possible zoonotic transmission to pigeon fanciers in six districts of Beijing where pigeon races are particularly popular. C. psittaci‐specific serum antibody titres were obtained from 370 pigeons and 79 fanciers using enzyme‐linked immunosorbent assay. In addition, 206 and 67 throat swabs were, respectively, collected from pigeons and fanciers and tested for the presence of chlamydial antigen using immunofluorescence. C. psittaci‐specific serum antibody was detected in 37 of 370 pigeons and 19 of 79 fanciers. Of 206 pigeon clinical specimens, 55 were positive for C. psittaci antigen, while 16 of 67 swabs from the pigeon fanciers were positive. Based on ompA sequence analysis, the genotype of several avian and human isolates was genotype B. Thus, both high‐titre C. psittaci‐specific antibody and C. psittaci‐specific antigen were found with relatively high frequency in the pigeon flocks as well as in the pigeon fanciers. Our study suggests that C. psittaci infection is prevalent among the racing pigeon population in Beijing. Moreover, detection of serum antibodies and antigen in pigeon fanciers suggests that exposure and possible zoonotic transmission of C. psittaci from racing pigeons to humans does occur. In view of the life‐threatening respiratory illness C. psittaci may cause in humans, regulatory public health measures, to prevent further spread of the pathogen in avian populations and possible transmission to exposed humans, are urgently needed.     

Compendium of measures to control Chlamydophila psittaci (formerly Chlamydia psittaci) infection among humans (psittacosis) and pet birds, 2005

Psittacosis, also known as parrot fever and ornithosis, is a bacterial infection of humans that can cause severe pneumonia and other serious health problems. It is caused by Chlamydophila psittaci, formerly known as Chlamydia psittaci. From 1988 through 2003, 935 human cases of psittacosis were reported to the CDC and most resulted from exposure to infected pet birds, usually cockatiels, parakeets, parrots, and macaws. In birds, C. psittaci infection is referred to as avian chlamydiosis. Infected birds shed the bacteria through feces and nasal discharges, and humans become infected from exposure to these materials. This compendium provides information about psittacosis and avian chlamydiosis to public health officials, physicians, veterinarians, the pet bird industry, and others concerned with controlling these diseases and protecting public health. The recommendations in this compendium provide standardized procedures for controlling avian chlamydiosis in birds, a vital step to protecting human health. This document will be reviewed and revised as necessary.     

Chlamydiosis in captive raptors

Chlamydia psittaci was isolated from four red-tailed hawks (Buteo jamaicensis) that died suddenly and from seven birds that survived at a raptor rehabilitation center in California in 1983. One hundred captive raptors representing 14 species in five families were subsequently tested serologically and by direct cloacal culture. C. psittaci was isolated from seven clinically normal birds. Forty-four percent of the raptors were considered positive using an enzyme-linked immunosorbent assay (ELISA), and 19% were suspects. The ELISA was repeated on 54 raptors in 1986. Forty-one percent of the birds were considered positive, and 35% were suspect, indicating that C. psittaci is endemic in the population.     

Bacteriological survey of feces from feral pigeons in Japan

Some public areas in Japan such as parks and gardens can be highly contaminated with pigeon feces. We examined levels of four bacterial contaminations in fecal samples from feral pigeons in 7 prefectures. We isolated Salmonella Typhimurium and S. Cerro from 17 (3.9%) of 436 samples, as well as Mycobacterium spp. including M. avium-intracellulare complex from 29 (19.0%) of 153 samples. The polymerase chain reaction detected Chlamydia psittaci and C. pecorum in 106 (22.9%) of 463 samples, but E. coli O-157 was not isolated from any of the samples. Our results indicate that pigeon feces are a source of several zoonotic agents for birds, animals and humans.     

Prevalence of Campylobacter jejuni in selected domestic and wild birds in Louisiana

Prevalence of Campylobacter jejuni was determined in a selected population of domestic and free-living birds submitted for necropsy to the Louisiana State Veterinary Medical Diagnostic Laboratory. The 445 cases examined included 13 orders of birds and yielded C. jejuni in 45 cases, representing an isolation rate of 10.1%. Prevalence was highest in Galliformes (25.2%), followed by Anseriformes (12.9%) and Columbiformes (8.3%). Only one isolation was made out of 179 Psittaciformes examined. Penner serotypes 1, 2, 4, and 16 were most commonly identified among the C. jejuni isolates. This study emphasizes the importance of Galliformes as reservoirs of C. jejuni. The commonality of these serotypes with isolates derived from humans suggests the zoonotic potential of Galliformes in relation to human campylobacteriosis. The isolation rate of 12.9% in Anseriformes implicates free-living and migratory waterfowl as carriers of C. jejuni. Results confirm that Psittaciformes represent a low risk of C. jejuni infection in humans.     

Methods of detection of Chlamydia psittaci in domesticated and wild birds

OBJECTIVE: To study the occurrence of Chlamydia psittaci in domesticated and wild birds and compare the sensitivity of molecular detection with cell culture isolation. DESIGN: Study of cell culture isolation and PCR detection of C psittaci in avian samples. PROCEDURE: Samples were obtained from 485 birds. Domesticated birds were selected at random from pet shops, private aviaries and zoos, while wild birds were captured locally, sampled, and immediately released. Swabs were collected from choanal slit, conjunctiva and cloaca of each bird and pooled. Samples were divided into equal portions for use in PCR dot-blot and cell culture detection. PCR and dot-blot detection was based on the ompB gene. RESULTS: Prevalence of infection varied markedly between flocks of captive birds. It was highest where there were frequent changes in the flock members or where there were many birds confined in small areas. C psittaci was not detected in wild birds or water birds. The sensitivity of cell culture compared to PCR dot-blot detection was 68%. All samples positive by cell culture were also positive by PCR. CONCLUSIONS: PCR-dot blot detection of C psittaci in birds appears to be more sensitive than cell culture isolation in this study. C psittaci infection of birds may occur in clinically normal captive birds.     

Antigenic analysis of avian Chlamydia psittaci using monoclonal antibodies to the major outer membrane protein.

Monoclonal antibodies to the major outer membrane protein (MOMP) of Chlamydia psittaci derived from a parrot were established for antigenic analysis of avian C. psittaci. With 17 monoclonal antibodies to MOMP, 17 reactivity patterns were identified on 112 strains of C. psittaci, C. pneumoniae and C. trachomatis, which were isolated from birds, mammals and humans in Japan, U.S.A., Canada and Taiwan, from 1938 to 1987. Immunological reactivity of budgerigar-derived strains to the monoclonal antibodies was different from that of pigeon-derived strains. Imported bird-derived strains were distinguishable from domestic bird-derived strains by the reactivity to the monoclonal antibodies. A close relationship between the subtypes and geographic origins was indicated on budgerigar-derived strains. On the contrary, various reactivity patterns were shown in pigeon-derived strains isolated in a narrow area. The monoclonal antibodies established in the present work may be useful probes for ecological study of avian C. psittaci.     

Health evaluation of free-ranging and captive blue-fronted Amazon parrots (Amazona aestiva) in the Gran chaco, Bolivia.

Bolivia has a total of 47 species of Psittacidae, seven of which have been identified in our study site, the semiarid Gran Chaco of the Isoso. One species, the blue-fronted parrot (Amazona aestiva), is frequently captured by local Isose?o Guaraní Indians for exploitation on the national and international market. These birds are often temporarily housed in small villages under unhygienic conditions with poultry and other domestic species. On occasion, these parrots escape back to the wild. Additionally, many of these birds are kept as pets or are used to lure wild. parrots within slingshot range for subsequent capture. In this study, we evaluated the health status, including the level of exposure to selected infectious agents, in the wild-caught captive birds and free-ranging birds. Physical examinations were performed, and blood was collected, from 54 live birds (20 captive and 34 free-ranging). Feces were collected from 15 birds (seven captive and eight free-ranging). Necropsies were also performed on four recently dead wild-caught birds. On serologic testing, no birds were found to have antibodies to avian influenza virus, Chlamydophila psittaci, infectious bronchitis virus, infectious bursal disease virus, infectious laryngotracheitis virus, Marek's disease virus, paramyxovirus-1, paramyxovirus-2, paramyxovirus-3, polyomavirus, eastern equine encephalitis virus, western equine encephalitis virus, or Venezuelan equine encephalitis virus. Positive antibody titers were found for psittacine herpesvirus (8/44, 18.2%), Aspergillus spp. (3/51, 5.9%), and Salmonella pullorum (33/49, 67.3%). All three of the birds that tested antibody positive for Aspergillus spp. were captive, whereas six of the eight and 15 of the 33 birds that tested positive for psittacine herpesvirus and S. pullorum, respectively, were wild.     

Detection of Chlamydophila psittaci by using SYBR green real-time PCR

Chlamydophila psittaci is the causative agent of human psittacosis and avian chlamydiosis. This zoonotic pathogen is frequently transmitted from infected birds to humans. Therefore proper and rapid detection of C. psittaci in birds is important to control this disease. We developed a method for detecting C. psittaci by using SYBR Green Real-time PCR based on targeting the cysteine-rich protein gene (envB) of C. psittaci. This one step procedure was highly sensitive and rapid for detection and quantification of C. psittaci from fecal samples. This assay was also able to detect other zoonotic Chlamydophila species such as C. abortus and C. felis. The assay is well suited for use as a routine detection method in veterinary medicine.     

Screening for several potential pathogens in feral pigeons (Columba livia) in Madrid

Background

Pathogens with the zoonotic potential to infect humans, such as Campylobacter jejuni, Campylobacter coli and Chlamydophila psittaci, can be found in feral pigeons (Columba livia). Given the high density of these birds in the public parks and gardens of most cities, they may pose a direct threat to public health.

Methods

A total of 118 pigeons were captured in three samplings carried out in 2006-2007 in public parks and gardens in Madrid, Spain. Standard haematological and morphological analyses were carried out on the pigeons. PCR was used to screen for the presence of Campylobacter jejuni, C. coli and Chlamydophila psittaci. Positive samples were confirmed by DNA sequencing.

Results

The analyses demonstrated a high prevalence of Chlamydophila psittaci (52.6%) and Campylobacter jejuni (69.1%) among the birds captured. In contrast, Campylobacter coli was rarely detected (1.1%).

Conclusions

Pigeons in Madrid can carry Chlamydophila psittaci and Campylobacter jejuni. They may be asymptomatic or subclinical carriers of both pathogens.