Seropositividad de Chlamydia psittaci en trabajadores expuestos a aves y revisión de la literatura: evidencia de circulación en Antioquia / Chlamydia psittaci seropositivity in workers exposed to birds and review of the literature: Evidence of circulation in Antioquia

Introducción. La psitacosis es una enfermedad zoonótica causada por Chlamydia psittaci. Esta bacteria es catalogada como un agente con potencial bioterrorista y ha causado múltiples brotes en trabajadores con exposición laboral a aves en diferentes lugares del mundo. En Colombia, no se hace seguimiento epidemiológico de la infección y existe una gran brecha en el conocimiento. Objetivos. Determinar la frecuencia de anticuerpos contra C. psittaci en trabajadores con exposición laboral a aves y sus factores asociados. Además, revisar la literatura en relación con los estudios sobre el tema realizados en Colombia. Materiales y métodos. Se llevó a cabo un estudio descriptivo, transversal, con intención analítica, en trabajadores en contacto con aves y se revisó la literatura científica relacionada en Colombia. Se detectaron anticuerpos IgM e IgG contra C. psittaci en suero por microinmunofluorescencia. La descripción de las características sociodemográficas y de exposición se hizo con frecuencias y medidas de resumen. Se exploraron factores asociados por análisis bivariados y multivariados. La revisión de la literatura científica y gris se hizo con búsqueda estructurada. Resultados. Se analizaron 54 trabajadores en contacto con aves y se encontró una prevalencia de anticuerpos del 31,5 %. El ejercer funciones de sacrificio y faenado de las aves sin ser médico veterinario fue un factor de riesgo para la presencia de anticuerpos. Solo se encontraron cuatro estudios previos sobre C. psittaci hechos en Colombia. Conclusiones. Este estudio constituye la primera evidencia de la circulación de C. psittaci en trabajadores en contacto con aves en Antioquia y el segundo reporte en el país. Estos hallazgos aportan desde la salud pública a la estrategia One Health.

Introduction. Psittacosis is a zoonotic disease caused by Chlamydia psittaci, a bacterium classified as an agent with bioterrorist potential. It has caused multiple outbreaks in exposed poultry workers around the world. Colombia has no epidemiological follow-up of the infection and a big knowledge gap. Objectives. To determine the antibodies' frequency against C. psittaci in workers with occupational exposure to birds and to review the literature on studies conducted in Colombia. Materials and methods. We conducted a cross-sectional descriptive study with analytical intent on workers in contact with birds and reviewed the related literature in Colombia. IgM and IgG serum antibodies against C. psittaci were detected by microimmunofluorescence. The sociodemographic and exposure characteristics were expressed as frequencies and summary measures. Associated factors were explored by bivariate and multivariate analysis. The scientific and gray literature review was done with a structured search. Results. We analyzed 54 workers in contact with birds. Antibody prevalence was 31.5%. Slaughtering and evisceration by non-veterinarians was a risk factor for antibody presence. There are only four previous studies on C. psittaci in Colombia. Conclusions. Here, we present the first evidence of C. psittaci circulation among workers exposed to birds in Antioquia and the second report in the country. These findings contribute to the "One Health" public health strategy.

Seropositividad a Chlamydophila psittaci en cotorras argentinas (Myiopsitta monachus) invasoras de la ciudad de Santiago de Chile

Introducción: Chlamydophila psittaci es una bacteria zoonótica e intracelular estricta, que provoca la psitacosis humana y su principal hospedero son las aves psitácidas. La cotorra argentina es un ave psitácida nativa de Sudamérica y actualmente considerada una especie invasora en 19 países, incluyendo Chile. Objetivo: Determinar positividad contra C. psittaci en muestras de suero y torulados de cotorras argentinas de vida libre capturadas en la Región Metropolitana de Chile. Métodos: Se analizaron 95 muestras de suero de pichones e individuos adultos de cotorras argentinas, a través de una prueba de ELISA indirecto utilizando un kit comercial. Posteriormente, se analizaron 40 tórulas nasotraqueales y cloacales de individuos adultos a través de una RPC en tiempo real específica para C. psittaci. Resultados: Se detectaron anticuerpos en muestras de suero de cinco individuos adultos de cotorras argentinas (n = 68), mientras que ninguno de los pichones analizados fue seropositivo (n = 27). Todas las muestras analizadas a través de RPC en tiempo real fueron negativas. Conclusión: Estos resultados demuestran por primera vez en Chile la exposición a C. psittaci en cotorras argentinas de vida libre, lo cual puede representar un riesgo importante para la transmisión de este patógeno a poblaciones humanas y animales.

Background: Chlamydophila psittaci is a zoonotic obligate intracellular bacterium that causes the human psittacosis, and its main host are psittacine birds. The monk parakeet is a psittacine bird native to South America, currently being considered an invasive species in 19 countries, including Chile. Aim: To determine positivity to C. psittaci in serum samples and swabs from free-ranging monk parakeets captured in the Metropolitan Region of Chile. Methods: Ninety-five serum samples from nestling chicks and adult monk parakeets were tested using an indirect ELISA test kit. Cloacal and nasotracheal swabs from 40 adult parakeets were further analyzed by C. psittaci-specific real-time PCR. Results: We found antibody titers in sera of five adult monk parakeets (n = 68) while none of the nestlings were seropositive (n = 27). All samples tested with real-time PCR were negative. Conclusions: Our results demónstrate for the first time in Chile the exposure to C. psittaci in free-ranging monk parakeets which may represent a significant risk of pathogen transmission to human and animal populations.

Type III secretory protein SINC of Chlamydia psittaci promotes host cell autophagy by activating the MAPK/ERK signaling pathway / 南方医科大学学报

OBJECTIVE@#To investigate the effects of SINC, a secreted protein of Chlamydia psittaci, on autophagy of host cells and the role of MAPK/ERK signaling pathway in mediating SINC-induced autophagy.@*METHODS@#RAW 264.7 cells treated with recombinant SINC were examined for changes in expression levels of LC3-II, Beclin-1, phosphorylated and total ERK1/2 using Western blotting. The expression level of LC3 in the treated cells was detected using immunofluorescence analysis, and the formation of autophagosomes and autolysosomes was observed with transmission electron microscopy (TEM). The effect of pretreatment with U0126 (a specific ERK inhibitor) on the expression levels of LC3-II and Beclin-1 in RAW 264.7 cells exposed to different concentrations of SINC was examined using Western blotting, and LC3 puncta in the cells was detected with immunofluorescence analysis.@*RESULTS@#The expression levels of LC3-II and Beclin-1 were the highest in RAW 264.7 cells treated with 2 μg/mL SINC for 12h. Immunofluorescence analysis showed exposure to SINC significantly increased the number of cells containing LC3 puncta, where the presence of autophagosomes and autolysosomes was detected. Exposure to 2 μg/mL SINC for 15 min resulted in the most significant increase of the ratios of p-ERK1/2/ERK1/2 in RAW 264.7 cells. Pretreatment of the cells with U0126 prior to SINC exposure significantly decreased the ratio of p-ERK1/2/ERK1/2, lowered the expression levels of LC3-II and Beclin-1, and decreased LC3 aggregation in the cells.@*CONCLUSIONS@#SINC exposure can induce autophagy in RAW 264.7 cells by activating the MAPK/ERK signaling pathway.

Molecular survey on zoonotic tick-borne bacteria and chlamydiae in feral pigeons (Columba livia domestica)

Objective: To determine the presence of zoonotic tick-borne bacteria in feral pigeons (Columba livia domestica) from urban areas. Methods: Spleen samples from 84 feral pigeons, found dead with traumatic injuries in urban areas, were examined by PCR to detect DNA of Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Rickettsia spp., and Chlamydophila spp. Results: Twenty (23.8%) pigeons were infected by tick-borne agents, in particular 2 (2.38%) animals resulted positive for Bartonella spp., 5 (5.95%) for C. burnetii, 5 (5.95%) for Rickettsia spp., 13 (15.47%) for B. burgdorferi sensu lato. All birds scored negative for A. phagocytophilum. Moreover, 17 (20.23%) pigeons were positive for Chlamydophila spp. and among them 10 (11.9%) for Chlamydophila psittaci. Mixed infections by two or three agents were detected in 8 (9.52%) animals. Conclusions: Feral pigeons living in urban and periurban areas are a hazard for the human health as source of several pathogens. The obtained results confirm pigeons as reservoirs of chlamydial agents and suggest that they may be involved in the epidemiology of zoonotic tick-borne infections too.

Molecular survey on zoonotic tick-borne bacteria and chlamydiae in feral pigeons (Columba livia domestica)

OBJECTIVE@#To determine the presence of zoonotic tick-borne bacteria in feral pigeons (Columba livia domestica) from urban areas.@*METHODS@#Spleen samples from 84 feral pigeons, found dead with traumatic injuries in urban areas, were examined by PCR to detect DNA of Anaplasma phagocytophilum, Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Rickettsia spp., and Chlamydophila spp.@*RESULTS@#Twenty (23.8%) pigeons were infected by tick-borne agents, in particular 2 (2.38%) animals resulted positive for Bartonella spp., 5 (5.95%) for C. burnetii, 5 (5.95%) for Rickettsia spp., 13 (15.47%) for B. burgdorferi sensu lato. All birds scored negative for A. phagocytophilum. Moreover, 17 (20.23%) pigeons were positive for Chlamydophila spp. and among them 10 (11.9%) for Chlamydophila psittaci. Mixed infections by two or three agents were detected in 8 (9.52%) animals.@*CONCLUSIONS@#Feral pigeons living in urban and periurban areas are a hazard for the human health as source of several pathogens. The obtained results confirm pigeons as reservoirs of chlamydial agents and suggest that they may be involved in the epidemiology of zoonotic tick-borne infections too.

Ocorrência de Chlamydophila psittaci em pombos (Columba livia) na cidade de Salvador, Bahia / Occurrence of the pigeon Chlamydophila psittaci in pigeons (Columba livia) within the city of Salvador, Bahia

A existência de populações numerosas de pombos (Columba livia) em centros urbanos, em quase todo o mundo, tem se tornado um risco à saúde pública em vista das zoonoses que podem transmitir. A infecção por Chlamydophila psittaci foi avaliada em pombos que frequentam áreas públicas, como praças, ruas e pontos turísticos na cidade de Salvador, Bahia, por meio da PCR em amostras de fezes frescas, suabes cloacais e orofaríngeos. O estudo revelou uma frequência de infecção por C. psittaci de 11,7% (16/137) dos pombos, e oito dos 10 locais pesquisados apresentavam aves infectadas. A detecção de C. psittaci em amostras de cloaca e orofaringe foi maior (15,8% - 3/19) que em amostras de fezes frescas (11% - 13/118). Os resultados demonstram a ocorrência de infecção por C. psittaci em pombos que habitam as áreas públicas da cidade de Salvador e apontam para a necessária elaboração de medidas de controle e monitoramento das populações de pombos urbanos, bem como de ações voltadas à conscientização da sociedade sobre os riscos à saúde pública.

The existence of numerous pigeon (Columba livia) populations in urban centers, in most of the world, has become a public-health risk given the zoonoses they can transmit. Infection with Chlamydophila psittaci was evaluated in pigeons that frequent public areas, such as squares, streets and tourist sites in the city of Salvador, through PCR from fresh feces samples, cloacal swabs and oropharyngeals. The study revealed a frequency of infection with C. psittaci of 11.7% (16/137) in pigeons, and infected birds were found in eight of the ten locales researched. The detection of C. psittaci in samples of cloaca and oropharyngs was greater (15.8% - 3/19) than in the samples of fresh feces (11% - 13/118). The results show the occurrence of infection with C. psittaciem in pigeons that live in public areas in the city of Salvador and points to the necessity of elaborating control and monitoring measures for the populations of urban pigeons as well as developing actions that will raise society´s awareness of these risks to public health.

Detection and characterization of chlamydophila psittaci in asymptomatic feral pigeons (Columba livia domestica) in central Thailand

Objective: To detect and characterize Chlamydophila psittaci (C. psittaci) in asymptomatic feral pigeons in central Thailand. Methods: A total 814 swabs from the trachea and cloacae of 407 non-clinical feral pigeons in central Thailand were collected and tested for the presence of C. psittaci. Results: A 10.8% of feral pigeons in the sample group were positive as determined by nested PCR primer specific to C. psittaci. The outer membrane protein A (ompA) gene of positive samples exhibited amino acid identity of C. psittaci ranging from 71 to 100% and were grouped in genotype B. Exceptionally, BF1676-56 isolate was closely related to Chlamydia avium with 99% identification of the 16S ribosomal (r) RNA gene. Conclusions: This is the first report on C. psittaci isolated from asymptomatic feral pigeons in Thailand, which provides knowledge for the disease status in pigeon populations in Thailand.

Detection and characterization ofChlamydophila psittaci in asymptomatic feral pigeons (Columba livia domestica) in central Thailand

Objective:To detect and characterizeChlamydophila psittaci(C. psittaci) in asymptomatic feral pigeons in centralThailand.Methods:A total814 swabs from the trachea and cloacae of407 non-clinical feral pigeons in centralThailand were collected and tested for the presence ofC. psittaci.Results:A10.8% of feral pigeons in the sample group were positive as determined by nestedPCR primer specific toC. psittaci.The outer membrane proteinA(ompA) gene of positive samples exhibited amino acid identity ofC. psittaci ranging from71 to100% and were grouped in genotypeB.Exceptionally,BF1676-56 isolate was closely related toChlamydia avium with 99% identification of the16S ribosomal(r)RNA gene.Conclusions:This is the first report onC. psittaci isolated from asymptomatic feral pigeons inThailand, which provides knowledge for the disease status in pigeon populations inThailand.

Detection and characterization of Chlamydophila psittaci in asymptomatic feral pigeons (Columba livia domestica) in central Thailand

OBJECTIVE@#To detect and characterize Chlamydophila psittaci (C. psittaci) in asymptomatic feral pigeons in central Thailand.@*METHODS@#A total 814 swabs from the trachea and cloacae of 407 non-clinical feral pigeons in central Thailand were collected and tested for the presence of C. psittaci.@*RESULTS@#A 10.8% of feral pigeons in the sample group were positive as determined by nested PCR primer specific to C. psittaci. The outer membrane protein A (ompA) gene of positive samples exhibited amino acid identity of C. psittaci ranging from 71 to 100% and were grouped in genotype B. Exceptionally, BF1676-56 isolate was closely related to Chlamydia avium with 99% identification of the 16S ribosomal (r) RNA gene.@*CONCLUSIONS@#This is the first report on C. psittaci isolated from asymptomatic feral pigeons in Thailand, which provides knowledge for the disease status in pigeon populations in Thailand.

Diagnóstico imuno-histoquímico e caracterização anatomopatológica de clamidiose em psitacídeos / Immunohistochemical diagnosis and pathological characterization of chlamydiosis in psittacine

A clamidiose é causada por Chlamydophila psittaci e representa uma das principais zoonoses de origem aviária. Realizou-se um estudo retrospectivo em psitacídeos do período de 1995 a 2012 e exame imuno-histoquímico (IHQ) anti-Chlamydia. Foram avaliados 111 casos, dos quais 12 foram a óbito devido à clamidiose. As aves eram provenientes de apreensão ou cativeiro (zoológicos, criatórios, centros de triagem e domicílios). À necropsia observou-se fígado aumentado (4/12) com áreas branco-amareladas (3/12), baço aumentado (2/12) e rompido (1/12), saco pericárdico com deposição de fibrina (1/12), polisserosite fibrinosa (1/12) e em três casos não havia lesões. Na avaliação histopatológica evidenciou-se hepatite necrótica mononuclear (7/12), hepatite mononuclear (3/12), hiperplasia de ductos biliares (8/12), esplenite necrótica histiocitária (9/12), hemossiderose em fígado (9/12) e baço (9/12), aerossaculite mononuclear (4/12), pericardite fibrino-heterofílica (2/12), necrose (1/12) e rarefação (1/12) linfoide de bursa de Fabricius, pneumonia fibrinosa (1/12), nefrite mononuclear (1/12) e granulomas renais (1/12). Observaram-se inclusões basofílicas intracitoplasmáticas (corpos elementares) em fígado (2/12), baço e rins (1/12). Evidenciou-se imunomarcação anti-Chlamydia em fígado (11/12), baço (7/9), pulmões (3/9), rins (2/8), intestinos (2/3), sacos aéreos (1/4) e bursa de Fabricius (1/2). A IHQ poderá ser utilizada como forma de diagnóstico definitivo post mortem de clamidiose em psitacídeos no Brasil...

Chlamydiosis is caused by Chlamydophila psittaci and is one of the most important avian zoonosis. A retrospective study in psittacines was performed from 1995 to 2012 with immunohistochemistry (IHC) anti-Chlamydia. Hundred eleven cases were evaluated and twelve birds died due to chlamydiosis. The birds were obtained from illegal commerce traffic or captive conditions (zoos, breeding birds, wildlife rehabilitation center and pets). Grossly, there were hepatomegaly (4/12) with yellowish-white areas (3/12), splenomegaly (2/12), splenic rupture (1/12), fibrin deposition in pericardial sac (1/12), fibrinous polyserositis (1/12), and in three cases lesion was not found. Histopathological evaluation revealed mononuclear necrotizing hepatitis (7/12), mononuclear hepatitis (3/12), biliary duct hyperplasia (8/12), histiocytic necrotizing splenitis (9/12), hemosiderosis in liver (9/12) and spleen (9/12), mononuclear aerosaculitis (4/12), fibrin heterophilic pericarditis (2/12), lymphoid necrosis (1/12) and depletion of bursa Fabricius (1/12), fibrinous pneumonia (1/12), mononuclear nephritis (1/12), and renal granulomas (1/12). Basophilic intracytoplasmic inclusions (elementary bodies) were observed in liver (2/12), spleen and kidney (1/12). Positive immunostaining for Chlamydia could be detected in liver (11/12), spleen (7/9), lung (3/9), kidney (2/8), intestines (2/3), air sacs (1/4) and bursa of Fabricius (1/2). It was concluded that IHC can be used as postmortem definitive diagnosis of chlamydiosis in psittacines...

Immunogenicity analysis of a recombinant protein CPSIT_p7 and its expression in HeLa cells during persistent Chlamydophila psittaci infection / 中华微生物学和免疫学杂志

Objective To construct a prokaryotic expression plasmid for CPSIT_p7 gene from Chlamydophila psittaci ( Cps) 6BC strain and to evaluate immunogenicity of the recombinant protein His-CPSIT_p7 and detect its dynamic expression at mRNA and protein levels in HeLa cells during persistent Cps infection.Methods The fusion protein His-CPSIT_p7 was expressed in E.coli BL21 and purified by Ni-NTA affinity chromatography .BALB/c mice were immunized with the recombinant protein to prepare polyclonal antibody for evaluation of the immunogenicity of His-CPSIT_p7 by ELISA.Penicillin sodium was used to establish a model of Cps persistence infection .RT-PCR and Western blot assay were performed to de-tect the expression of CPSIT_p7 at mRNA and protein levels during Cps persistent infection .Results The fusion protein His-CPSIT_p7 was successfully expressed with the use of constructed recombinant expression plasmid pET30 a-CPSIT_p7 and purified .ELISA result showed that the specific antibody titer against CPSIT_p7 reached 1 ∶1 000 000 on the 40th days after immunization .The expression of CPSIT_p7 at mRNA and protein levels were increased in a time-dependent manner in Cps-infected HeLa cells .The peak of mRNA level was reached at the time point of 36 hours after infection , followed by a time-dependent decrease during Cps acute infection .However , the expression of CPSIT_p7 at mRNA and protein levels were not decreased until 60 hours after infection during Cps persistent infection .Conclusion His-CPSIT_p7 protein was suc-cessfully expressed in the prokaryotic expression system and purified , showing an advantage of good immuno-genicity.Highly expressed CPSIT_p7 at mRNA and protein levels were detected during Cps persistent infection.

Localization and temporal expression of CPSIT_0271 in Chlamydia psittaci-infected cells and its stimula-tory effects on the expression of proinflammatory cytokines by THP-1 cells / 中华微生物学和免疫学杂志

Objective To study the intracellular localization and temporal expression of CPSIT_0271 in Chlamydia psittaci-infected cells; and to investigate the effects of recombinant GST-CPSIT_0271 protein on the expression of proinflammatory cytokines including IL-6, IL-1βand TNF-αby THP-1 cells.Methods The gene encoding CPSIT_0271 of Chlamydia psittaci was expressed as fusion protein ( GST-CPSIT_0271 ) in E.coli.The polyclonal antibody was prepared by immunizing BALB /c mice with the purified recombinant pro-tein.Antibody titer was determined by ELISA .Indirect immunofluorescence assay ( IFA) was performed to lo-cate the endogenous CPSIT_0271 protein in C.psittaci-infected cells .The expression characteristics of CPSIT_0271 protein were detected by Western blot in C.psittaci-infected HeLa cells at different time points .The levels of IL-6, IL-1βand TNF-αwere analyzed by ELISA after stimulating THP-1 cells with different concentrations of CPSIT_0271 protein.Results CPSIT_0271 protein was found to express in the chlamydia inclusion of C.psittaci-infected HeLa cells .The expression of CPSIT_0271 protein was detected firstly at 36 h and increased at 48 h after C.psittaci infection.The titer of anti-CPSIT_0271 specific antibody in GST-CPSIT_0271 immu-nized mice reached to 1 ∶16 000.GST-CPSIT_0271 protein increased the levels of IL-6, IL-1βand TNF-αin THP-1 cells in a dose-dependent manner in the range of 2 to 5 μg/ml.The levels of TNF-αand IL-1βreached their peaks at 24 h, and IL-6 level peaked at 48 h upon the stimulation by 5 μg/ml of GST-CPSIT_0271 pro-tein.Conclusion CPSIT_0271 expressed in inclusion bodies of Chlamydia psittaci in the infected cells , sug-gesting it might be a late expression gene .GST-CPSIT_0271 protein shows good immunogenicity and enhances the expressions of IL-6, IL-1βand TNF-αin THP-1 cells.

Clinico-pathological Features of Chlamydophila psittaci Infection in Parrots and Genetic Characterization of the Isolates / 대한수의학회지

Avian chlamydiosis is caused by Chlamydophila psittaci and considered as one of an important zoonotic disease throughout the world. Among more than 400 avian species including poultry and pet birds susceptible to the disease, psittacine birds were known to be mostly susceptible hosts. In Korea, no outbreak of the disease and genetic analysis of the agent in poultry and pet birds have been reported. With histopathological findings and genetic identification of a causative agent, avian chlamydiosis was identified in parrots submitted from the same pet bird farm in 2006 and 2009 for the diagnosis. Based on genetic sequences and phylogenetic analysis of ompA gene, the two isolates of Chlamydophila psittaci showed 100% of genetic similarity and belonged to genotype A, suggesting that the same agent might be continuously circulated in the farm. This result indicates that serological survey of the disease in pet bird farms and impact of the disease on significance in public health may be further studied.

Detección de anticuerpos IgG contra Chlamydophila psittaci en aves psitácidas en cautiverio: Maracay, Venezuela / Detection of IgG antibodies against Chlamydophila psittaci in captive psittacine birds: Maracay, Venezuela

La clamidiosis es una enfermedad cosmopolita muy conocida en aves de la familia Psitacidae, cuyo agente etiológico es Chlamydophila psittaci. Puede causar desde una leve neumonía hasta la muerte del animal; además es posible su transmisión a otros animales y al humano a través de la inhalación del microorganismo, produciendo cuadros severos de neumonía y otras patologías. Las poblaciones con mayor riesgo de adquirir esta zoonosis son veterinarios, criadores de aves y toda persona en contacto con aves infectadas. En Venezuela no existen registros epidemiológicos de esta enfermedad en psitácidos ni en humanos. El objetivo de este trabajo fue detectar la presencia de anticuerpos contra Chlamydophila psittaci en aves psitácidas del parque Zoológico Las Delicias y el Zoocriadero Canaima por medio de la técnica de ELISA en fase sólida. Para ello se analizaron 24 muestras de suero de aves encontrándose 17 positivas, lo que se traduce en una prevalencia de anticuerpos IgG contra Cp. psittaci de 71%. Con estos resultados se pretende llamar la atención de las autoridades sanitarias respectivas sobre el impacto e importancia de la enfermedad en Venezuela.

Chlamydiosis is a well known cosmopolitan disease in birds belonging to the Psittacidae family whose etiologic agent is Chlamydophila psittaci. It can produce from mild pneumonia to the death of the animal; its transmission to other animals and to humans is also possible through the inhalation of the microorganism, producing severe pneumonia episodes, as well as other pathologies. The populations with the highest risk of acquiring this zoonosis are those of veterinaries, bird keepers and all persons in contact with infected birds. In Venezuela there are no epidemiologic registries of this disease in psittacine birds or in humans. The purpose of this study was to detect the presence of antibodies against Chlamydophila psittaci in psittacine birds from the Las Delicias zoo and the Canaima Zoogrower through a solid phase ELISA technique. Twenty-four serum samples taken from birds were analyzed and 17 were found to be positive, which results in a 71% prevalence of IgG antibodies against C. psittaci. With these results we intend to call the attention of the respective public health authorities on the impact and importance of this disease in Venezuela.

Infecção por C. psittaci: uma revisão com ênfase em psitacídeos / C. psittaci infection: a review with emphasis in psittacines

A clamidiose ou ornitose é uma doença infecciosa, causada pela bactéria Chlamydophila psittaci, que acomete aves e mamíferos. Trata-se de uma das principais zoonoses de origem aviária. A transmissão ocorre principalmente por inalação de secreções contaminadas. Os sinais clínicos mais comuns incluem alterações no sistema gastrointestinal, respiratório e ocular, porém é possível encontrar aves infectadas sem sinais aparentes, dificultando a identificação da doença. O diagnóstico definitivo em aves vivas pode ser difícil, devido às características da infecção pela bactéria. Há duas principais abordagens para o diagnóstico, a primeira envolve a detecção direta da bactéria e a segunda implica a detecção de anticorpos anti-Chlamydophila sp. O tratamento é longo e envolve o uso de tetraciclinas, quinolonas ou macrolídeos, durante 21-45 dias, dependendo da espécie e do fármaco de escolha. Atualmente, o Brasil não dispõe de medidas padronizadas que visam a guiar o clínico na identificação, manejo e tratamento para a doença. Tais medidas tornam-se necessárias, bem como a pesquisa de novos métodos diagnósticos e auxiliares para a doença.

Chlamydiosis or ornitosis is an infectious disease which affects birds and mammals caused by the bacteria Chlamydophila psittaci. It is one of the most important avian zoonosis. The transmission occurs through inhalation of infected secretions. The most common clinical signs include problems in the gastrointestinal, respiratory and ocular tracts. However, it is possible to find infected birds with no clinical signs, which hinders the diagnosis. Definitive diagnosis in live birds can be difficult, because of the bacteria's infection characteristic. There are two main approaches to the diagnosis, the first one involves the direct detection of the bacteria, the second one involves the detection of antibodies anti-Chlamydophila sp. The treatment is long and includes the use of tetraciclines, quinolones and macrolides, during 21-45 days, depending of the specie or drug of choice. Currently, Brazil has no standardized procedures to guide clinicians in the identification, management, and treatment of the disease. Such measures become necessary, as well as research on new diagnostic methods.

Primera evidencia de circulación de Chlamydophila psittaci en Colombia: posible riesgo de salud pública / First evidence of Chlamydophila psittaci circulation in Colombia: a possible public health risk

Objetivo Establecer la seroprevalencia de Chlamydophila psittaci en aves del género Amazona spp y en trabajadores de algunos zoológicos y CAV (centros de atención y valoración de fauna silvestre). Metodología Se analizaron 138 sueros de aves del género Amazona spp, 24 sueros de otras especies de aves y 39 sueros humanos por ELISA indirecta. Se utilizó el antígeno RMOMP (major outer membrane protein of Chlamydophila psittaci). Para el conjugado de aves se utilizo una anti-IgG de turkey-chicken marcado con biotina para el conjugado humano se utilizo una anti-IgG marcada con peroxidasa. Los sueros fueron diluidos 1:100. Resultados De los 138 sueros de aves del género Amazona spp 118 (85 por ciento) resultaron positivos. La seroprevalencia por región fue la siguiente: CAV Torre cuatro de Caldas 36 (90 por ciento), Zoológico de Barranquilla 14 (87 por ciento), CAV Montería 28 (85 por ciento), Zoológico de Cali 21 (84 por ciento) y CAV Victoria del Oriente Caldense 19 (79 por ciento). En humanos la seroprevalencia total fue del 78 por ciento (30/39) la distribución fue la siguiente: CAV Montería 9 trabajadores (100 por ciento), Zoológico de Barranquilla 9 (90 por ciento), CAV Torre 4 de Caldas 4 trabajadores (80 por ciento), CAV Victoria del Oriente Caldense 3 (75 por ciento) y 5 (45 por ciento) trabajadores del Zoológico de Cali. Conclusiones La alta seroprevalencia de Chlamydophila psittaci en aves (86,4 por ciento) y humanos (78 por ciento) representa la primera evidencia de la circulación de este microorganismo en Colombia, la presencia del agente etiologico de la ornitosis podría representar un riesgo de salud publica.

Objective To establish the seroprevalence of Chlamydophila psittaci in birds of Amazona spp genus and workers from some zoos and CAVs (centers for attention and evaluation of wildlife) of Colombia. Methods We analyzed 138 sera from birds of the genus Amazona spp, 24 sera from other species of birds and 39 human sera by indirect ELISA. RMOMP was used as antigen (major outer membrane protein of Chlamydophila psittaci). For the conjugate of birds it was used an anti-turkey-chicken IgG labeled with biotin, for the human conjugate we used an anti-IgG labeled with peroxidase. Sera were diluted 1:100. Results Of the 138 sera from birds of the genus Amazona spp 118 (85 percent) were seropositive. Regional seroprevalence was as follow: Caldas CAV Torre cuatro 36 (90 percent), CAVâs Monteria 28 (85 percent), Barranquillaâs Zoo 14 (87 percent), Caliâs Zoo 21 (84 percent) and from the CAV Victoria del Oriente Caldense 19 (79 percent) sera were seropositive. Regarding seroprevalence in humans, 30 of the 39 (78 percent) were seropositive, regional seroprevalence was as follow: Montería 9 (100 percent) workers, Barranquillaâs Zoo 9 (90 percent), CAV Caldas Tower four 4 (80 percent), Caliâs Zoo 5 (45 percent) and CAV Caldense Victoria del oriente 3 (75 percent) were seropositive. Conclusions The high seroprevalence of Chlamydophila psittaci in birds (86,4 percent) and humans (78 percent) showed the first evidence of circulation of this microorganism in Colombia; the circulation of the etiological agent of psittacosis may represent a public health risk.

Study on the diagnosis application using recombinant protein of immunodominant region gene of chlamydial protease-like activity factor from Chlamydophila psittaci / 中华微生物学和免疫学杂志

Objective To clone and express the immunodominant domain of the chlamydial proteaselike activity factor(CPAF) from Chlamydophila psittaci(Cps) and evaluated the diagnosing value of the recombinant protein in Cps infection.Methods The immunodominant region epitope of CPAF (CPAFm,A196-A450)from Cps was chosen according to bioinformatics analysis and references.The specific primer was designed and the gene was amplified by PCR and then ligated into a pGEX6p-2 vector.Recombinant protein was induced to express by IPTG and analyzed by SDS-PAGE and Western blot.Indirect EL1SA method of serological diagnosis was established with the reorganization protein as coating antigen.One hundred and eighty sera samples from ducks with respiratory tract infection symptoms were detected with the established indirect ELISA and a commercial ELISA-kit to assess the value of the recombinant protein in serodiagnosis.The results were further identified with Western blot.Results Prokaryotic expression vector pGEX6p-2/CPAFm was constructed and a 54x103 fusion protein was attained.The indirect ELISA method was established with reorganization protein for envelope antigen.Using the indirect ELISA to detect Cps lgG positive and negative reference sera,the sensitivity and specificity were both 100％ (20/20).And the recombinant protein has no cross reaction with either Chlamydophila pneumoniae or Chlamydophila trachomatis.The concordance rate between the indirect ELISA and Western blot to 180 ducks sera samples was 100％,while the concordance rate of the commercial ELISA kit was 77.5％-95.0％.Conclusion The prepared recombinant protein of the CPAF immunodominant region epitope gene from Gps can highly benefit on developing new indirect ELISA as methods to detect specific anti-Cps antibodies.

Evidence of zoonotic Chlamydophila psittaci transmission in a population at risk in Zulia state, Venezuela / Evidencia de transmisión zoonótica de Chlamydophila psittacci en una población de riesgo del Estado Zulia, Venezuela

Objective This study was aimed at investigating the frequency of infection by Cp. psittaci and determining its genotype in individuals at potential risk of exposure to the bacteria. Methodology The study involved 170 individuals: a risk group (n= 96) and a low-risk control group (n=74). Cp. psittaci was detected and genotyped by single-tube nested PCR and ompA gene sequencing. Results Eight (8.3 %) positive cases were detected in the risk group and 1 (1.4 %) in the control group (p<0.04). Cp. psittaci was found in 16.7 % of pigeons' fecal samples. Cp. psittaci infection with was more frequent in symptomatic (17.7 %) than asymptomatic (6.3 %) individuals in the risk group. Analysing the genomes isolated from human and bird specimens revealed the presence of genotype B. Conclusion The presence of Cp. psittaci genotype B in the population being evaluated could have been attributed to zoonotic transmission from pigeons to humans, an underestimated potential public health problem in Venezuela requiring the health authorities' involvement.

Objetivo El objetivo de este estudio fue investigar la frecuencia de infecciones por Cp. psittaci y determinar su genotipo en individuos con potencial riesgo de exposición a la bacteria. Metodología Se incluyeron 170 individuos, un grupo de riesgo (n=96) y un grupo control (n=74). La detección y genotipificación de Cp. psittaci se llevó a cabo por PCR anidada y secuenciación del gen ompA. Resultados Se detectaron ocho (8,3 %) casos positivos en el grupo de riesgo y 1 (1,35 %) en el grupo control (p<0,04). Cp. psittaci fue detectada en 16,7 % muestras fecales de palomas. En el grupo de riesgo, la frecuencia de infección por Cp. psittaci fue 17,7 % en individuos sintomáticos y 6,3% en asintomáticos. El análisis de los genomas aislados de muestras humanas y aves, revelaron la presencia del genotipo B. Conclusión La presencia de Cp. psittaci genotipo B en la población evaluada podría ser atribuida a transmisión zoonótica de palomas a humanos, un potencial problema de salud pública en nuestra región que requiere la intervención de autoridades sanitarias.

A Case of Keratoconjunctivitis Caused by Chlamydophila Psittaci

PURPOSE: Only a few cases of keratoconjunctivitis caused by Chlamydophila psittaci have been reported worldwide, and no case reported in Korea. We report an atypical case of keratoconjunctivitis caused by Chlamydophila psittaci. CASE SUMMARY: A 34-year-old male patient who had raised a parrot at home presented with three weeks of conjunctival injection and a week of ocular pain in his left eye. There were papillae on the left upper and lower tarsal conjunctiva and punctuate epithelial erosion of the entire cornea. He also complained of dizziness, fever, and dyspnea. Upon chest X-ray, consolidation on the right middle lobe was apparent. The Chlamydophila IgM antibody test was positive, and the pneumonia improved quickly. Nevertheless, signs of keratoconjunctivitis persisted despite 3-week treatment with oral doxycycline. As a result, the patient received an additional 10-day treatment with oral azithromycin. Four weeks after the first visit, symptoms were improving gradually, and, after six weeks, no signs of keratoconjunctivitis remained except minimal erosion. CONCLUSIONS: When patients show keratoconjunctivitis after contact with a bird, prolonged ketatoconjunctivitis by Chlamydophila psittaci should be considered.

Comparison on two pairs of primer used in fluorescent quantitative assay to detect genes encoding the major outer membrane protein and the polymorphic membrane protein of Chlamydophila psittaci / 中国人兽共患病学报

Two pairs of primer targeting to the genes encoding the major outer membrane protein (MOMP) and the polymorphic membrane protein (PMP) of Chlamydophila psittaci (Cps) designed and used in the fluorescent quantitative PCR assay in the detection of this microorganism were compared in the their sensitivity and specificity.It was demonstrated that both pairs of primer could be used successfully to detect the presence of Cps of the epidemic strains isolated in China.The specificity of PMP primer used was higher than that of MOMP primer. In case of high target concentration in samples, the sensitivity of the former was also superior to that of the latter, but the amplification efficiency of the former was lower than that of the latter. The MOMP primer seemed to be more suitable to detect Cps by using the real-time quantitative assay.As demonstrated by the real-time quantitative PCR assay, great difference in the genomic copy numbers of the PMP gene existed in the epidemic strains of Cps isolated in China.