Development and comparative evaluation of droplet digital PCR and quantitative PCR for the detection and quantification of Chlamydia psittaci.

Chlamydia psittaci is a zoonotic pathogen mainly transmitted by psittacine birds and poultry. The low shedding rate of the pathogen in the apparently healthy birds and human clinical cases may result in false-negative results. In the present study, a droplet digital PCR (ddPCR) assay was developed and compared with optimized quantitative PCR (qPCR) for the detection of C. psittaci from the clinical samples. The ddPCR assay was found to be comparatively more sensitive than the qPCR, wherein the limit of detection (LOD) of ddPCR was upto 2.4 copies of the DNA template, whereas, the qPCR could detect upto 38 copies of the DNA template in the reaction mixture. Overall, the developed ddPCR assay was found to be robust, specific, and could reliably quantify up to 17.8 copies of the DNA template. Finally, the applicability of the developed ddPCR assay was tested by screening the field samples (n = 124), comprising lung tissues from dead poultry and feral birds; pooled faecal samples from the free-living birds, commercial and backyard poultry farms; pharyngeal and cloacal swabs collected from the duck farms. Of these, a total of seven samples were found to be positive by the ddPCR, whereas, three samples could be detected as positive using the qPCR. The developed ddPCR could serve as a reliable screening tool, particularly in those clinical samples wherein the shedding of C. psittaci is substantially very low.

Real-time fluorometric and end-point colorimetric isothermal assays for detection of equine pathogens C. psittaci and equine herpes virus 1: validation, comparison and application at the point of care.

BACKGROUND: C. psittaci has recently emerged as an equine abortigenic pathogen causing significant losses to the Australian Thoroughbred industry, while Equine herpesvirus-1 (EHV-1) is a well-recognized abortigenic agent. Diagnosis of these agents is based on molecular assays in diagnostic laboratories. In this study, we validated C. psittaci and newly developed EHV-1 Loop Mediated Isothermal Amplification (LAMP) assays performed in a real-time fluorometer (rtLAMP) against the reference diagnostic assays. We also evaluated isothermal amplification using commercially available colorimetric mix (cLAMP), and SYBR Green DNA binding dye (sgLAMP) for "naked eye" end-point detection when testing 'real-world' clinical samples. Finally, we applied the C. psittaci LAMP assays in two pilot Point-of-Care (POC) studies in an equine hospital. RESULTS: The analytical sensitivity of C. psittaci and EHV-1 rt-, and colorimetric LAMPs was determined as one and 10 genome equivalents per reaction, respectively. Compared to reference diagnostic qPCR assays, the C. psittaci rtLAMP showed sensitivity of 100%, specificity of 97.5, and 98.86% agreement, while EHV-1 rtLAMP showed 86.96% sensitivity, 100% specificity, and 91.43% agreement. When testing rapidly processed clinical samples, all three C. psittaci rt-, c-, sg-LAMP assays were highly congruent with each other, with Kappa values of 0. 906 for sgLAMP and 0. 821 for cLAMP when compared to rtLAMP. EHV-1 testing also revealed high congruence between the assays, with Kappa values of 0.784 for cLAMP and 0.638 for sgLAMP when compared to rtLAMP. The congruence between LAMP assays and the C. psittaci or EHV-1 qPCR assays was high, with agreements ranging from 94.12 to 100% for C. psittaci, and 88.24 to 94.12% for EHV-1, respectively. At the POC, the C. psittaci rt- and c-LAMP assays using rapidly processed swabs were performed by technicians with no prior molecular experience, and the overall congruence between the POC C. psittaci LAMPs and the qPCR assays ranged between 90.91-100%. CONCLUSIONS: This study describes reliable POC options for the detection of the equine pathogens: C. psittaci and EHV-1. Testing 'real-world' samples in equine clinical setting, represents a proof-of-concept that POC isothermal diagnostics can be applied to rapid disease screening in the equine industry.

Investigation of chlamydophilosis from naturally infected cats.

BACKGROUND: Chlamydophila felis, formerly known as Chlamydia psittaci var. felis, is frequently associated with ocular, respiratory, and occasionally reproduction tract infections. Even though the infection is sometimes asymptomatic, it potentially results in a latent immunosuppressive infection. OBJECTIVE: This study aimed to identify occurrences of feline chlamydophilosis, rarely reported in cats in Indonesia. METHODS: The observation was conducted in three cats with clinical signs of Cp. felis infection, particularly relapsing conjunctivitis. The cats' histories were recorded based on owners' information. Conjunctival swabs were sampled for cytology examination and molecular assay detection. A phylogenetic tree was generated using MEGA-X software to reveal group clustering. A post-mortem examination was performed on the cat that died during an examination. RESULTS: Cp. felis was detected in both cytological examination and polymerase chain reaction assay. The phylogenetic tree demonstrated that the Cp. felis isolated in this study clustered with several other isolates from the other countries. Cp. felis can be isolated from cats with different clinical manifestations and levels of severity. The chronic fatal infection demonstrated interstitial broncho-pneumonia under histopathological examination. CONCLUSIONS: Molecular assay of Cp. felis is always recommended to obtain a definitive diagnosis of feline chlamydophilosis since the disease can have various clinical manifestations. Even though it may be subclinical and is often not fatal, an infected cat may be a carrier that could spread the pathogen in the surrounding environment. Serious disease management is suggested to avoid high costs associated with regularly relapsing disease.

Detection of Chlamydia psittaci in both blood and bronchoalveolar lavage fluid using metagenomic next-generation sequencing: A case report.

RATIONALE: Chlamydia psittaci (C psittaci) is a gram-negative obligate intracellular parasite, with birds as main hosts. The main route of infection in humans is inhalation of aerosols from contaminated animal excreta through the respiratory tract. The main manifestation of C psittaci infection is pneumonia. Patients suffering from severe infection are prone to sepsis and multiple organ failure. We report a case of simultaneous detection of C psittaci in blood and bronchoalveolar lavage fluid using metagenomic next-generation sequencing (mNGS) technology. PATIENT CONCERNS: The 71-year-old male patient was a farmer with a long history of raising poultry and initial symptoms of fever and muscle pain accompanied by limb weakness and paroxysmal cough. DIAGNOSES: The patient was diagnosed with sepsis, severe pneumonia, and multiple organ failure. INTERVENTIONS: Anti-infective therapy with doxycycline and meropenem was applied. OUTCOMES: The patient's body temperature and infection indicators improved and the chest X-ray findings showed the amelioration of lesions after 18 days of treatment. The patient was discharged without treatment on hospital day 19 due to financial constraints and subsequently died after 7 days. LESSONS: mNGS is an excellent diagnostic tool when specific pathogens are undetected by traditional assays.

A case of chlamydia psittaci caused severe pneumonia and meningitis diagnosed by metagenome next-generation sequencing and clinical analysis: a case report and literature review.

BACKGROUND: Psittacosis, which is also known as parrot fever, is Chlamydia psittaci (C. psittaci) caused infectious disease. The clinical manifestations vary from asymptomatic infection to severe atypical pneumonia or even fatal meningitis. Early recognition of psittacosis is difficult because of its nonspecific clinical manifestations. Culture and gene probe techniques for C. psittaci are not available for routine clinical use, which makes the diagnosis difficult too. Although psittacosis has increasingly been recognized and reported in recent years, cure of severe pneumonia complicated with meningitis, with etiologic diagnosis aided by the use of metagenomic next-generation sequencing (mNGS), is still uncommon. So, it is necessary to report and review such potentially fatal case. CASE PRESENTATION: This report describes a 54-year-old woman with C. psittaci caused severe atypical pneumonia and meningitis. She presented with symptoms of fever, dry cough and dyspnea, accompanied by prominent headache. Her condition deteriorated rapidly to respiratory failure and lethargy under the treatment of empirical antibacterial agents, and was treated with invasive mechanical ventilation soon. She denied contact with birds, poultry or horses, but unbiased mNGS of both the bronchoalveolar lavage fluid (BALF) and the cerebrospinal fluid (CSF) identified sequence reads corresponding to C. psittaci infection, and there was no sequence read corresponding to other probable pathogens. Combined use of targeted antimicrobial agents of tetracyclines, macrolides and fluoroquinolones was carried out, and the patient's condition improved and she was discharged home 28 days later. Her status returned close to premorbid condition on day 60 of follow-up. CONCLUSIONS: When clinicians come across a patient with atypical pneumonia accompanied by symptoms of meningitis, psittacosis should be taken into consideration. mNGS is a promising detection method in such condition and is recommended.

Epidemiology of Chlamydia psittaci infections in pregnant Thoroughbred mares and foals.

Late-term foal loss due to the traditional avian pathogen Chlamydia psittaci recently emerged as a threat to the Australian Thoroughbred industry. A longitudinal study of 14 stud farms was undertaken to better understand C. psittaci infection in pregnant mares and their foals by evaluating C. psittaci prevalence, equine herpesvirus-1 (EHV-1) co-infection, avian reservoirs, and potential risk factors. Mucosal swabs taken from 228 healthy pregnant mares and their foals were tested for C. psittaci and EHV-1 using species-specific qPCR assays. No foal loss was recorded due to either pathogen, and no mare tested positive to either C. psittaci or EHV-1. However, healthy newborn foals tested positive to both pathogens, at low levels, with 13.2% (n = 30/228) and 14.5% (n = 33/228) prevalence for C. psittaci and EHV-1, respectively. Co-infection occurred in 1.3% (n = 3/228) of foals. In avian environmental faecal samples collected from the same studs, C. psittaci was detected at 5.3% (n = 5/94). Multiple logistic regression modelling found that foals born in winter were more likely to be infected with C. psittaci (adjusted odds ratio = 15.83; P < 0.001; Confidence Interval 5.12-48.49). Being a maiden mare, absence of prophylactic vaginal suture, interventions in the last trimester and residing on a farm with prior history of C. psittaci abortion posed no higher risk to infection in the newborn. Analysis of all reported C. psittaci abortion cases (Hunter Valley, 2016-2019) revealed a dominant C. psittaci sequence type (denoted ST24) and a significant correlation with frost events (Spearmans' rho = 0.44; P = 0.002).

Metagenomic next-generation sequencing in the family outbreak of psittacosis: the first reported family outbreak of psittacosis in China under COVID-19.

Chlamydia psittaci infection in humans, also known as psittacosis, is usually believed to be an uncommon disease which mainly presents as community-acquired pneumonia (CAP). It is usually sporadic, but outbreaks of infection may occasionally occur. In outbreaks, diagnosis and investigations were usually hampered by the non-specificity of laboratory testing methods to identify C. psittaci. In this study, we use metagenomic next-generation sequencing (mNGS) in the diagnosis of a family outbreak of psittacosis under COVID-19. Three members of an extended family of 6 persons developed psittacosis with pneumonia and hepatic involvement with common symptoms of fever and weakness. Two newly purchased pet parrots, which had died successively, were probably the primary source of infection. Imagings show lung consolidations and infiltrates, which are difficult to be differentiated from CAP caused by other common pathogens. mNGS rapidly identified the infecting agent as C. psittaci within 48 h. The results of this work suggest that there are not characteristic clinical manifestations and imagings of psittacosis pneumonia which can differentiate from CAP caused by other pathogens. The use of mNGS can improve accuracy and reduce the delay in the diagnosis of psittacosis especially during the outbreak, which can shorten the course of the disease control. Family outbreak under COVID-19 may be related to the familial aggregation due to the epidemic. To our knowledge, this is the first reported family outbreak of psittacosis in China, and the first reported psittacosis outbreak identified by the method of mNGS in the world.

Use of Real-Time PCR for Chlamydia psittaci Detection in Human Specimens During an Outbreak of Psittacosis - Georgia and Virginia, 2018.

Psittacosis is typically a mild febrile respiratory illness caused by infection with the bacterium Chlamydia psittaci and usually transmitted to humans by infected birds (1). On average, 11 psittacosis cases per year were reported in the United States during 2000-2017. During August-October 2018, the largest U.S. psittacosis outbreak in 30 years (82 cases identified*) occurred in two poultry slaughter plants, one each in Virginia and Georgia, that shared source farms (2). CDC used C. psittaci real-time polymerase chain reaction (PCR) to test 54 human specimens from this outbreak. This was the largest number of human specimens from a single outbreak ever tested for C. psittaci using real-time PCR, which is faster and more sensitive than commercially available serologic tests. This represented a rare opportunity to assess the utility of multiple specimen types for real-time PCR detection of C. psittaci. C. psittaci was detected more frequently in lower respiratory specimens (59% [10 of 17]) and stool (four of five) than in upper respiratory specimens (7% [two of 28]). Among six patients with sputum and nasopharyngeal swabs tested, C. psittaci was detected only in sputum in five patients. Cycle threshold (Ct) values suggested bacterial load was higher in lower respiratory specimens than in nasopharyngeal swabs. These findings support prioritizing lower respiratory specimens for real-time PCR detection of C. psittaci. Stool specimens might also have utility for diagnosis of psittacosis.

Development and evaluation of a novel quenching probe PCR (GENECUBE) assay for rapidly detecting and distinguishing between Chlamydia pneumoniae and Chlamydia psittaci.

Early detection of the family Chlamydiaceae as pathogens is essential worldwide for the rapid and sufficient management of atypical pneumonia. GENECUBE (TOYOBO) is a novel fully automated gene analyzer capable of amplifying and detecting target DNAs within 50 min. In this study, we developed a new PCR assay with a specific quenching probe (PCR-QC assay) for rapidly distinguishing between Chlamydia pneumoniae (CPN) and Chlamydia psittaci (CPS). The PCR-QC assay enabled us to precisely and simultaneously detect the 2 different types of DNA fragments even in a mixed sample by identifying unique melting temperatures. Next, we examined a total of 300 frozen samples from patients with respiratory tract infection using the PCR-QC assay and the cell culture method as the gold standard. Kappa index for agreement between the PCR-QC assay and the culture method was 0.43 (95% confidential interval (CI): 0.08-0.78). The sensitivity and specificity of the PCR-QC assay were 36.3% (4/11; 95% CI: 10.9-69.2%)) and 99.0% (286/289; 95% CI: 97.0-99.8%), respectively. The samples positive for CPN (n = 13) or CPS (n = 1) by either method were also examined by a conventional PCR TaqMan assay, which produced the same results as those from the PCR-QC assay. Furthermore, the PCR-QC assay using GENECUBE shortened the full detection time for CPN or CPS (within 50 min vs. more than 2 to 3 h) compared with conventional PCR TaqMan assays. Therefore, the new PCR-QC assay system equipped with GENECUBE is useful for rapidly detecting CPN or CPS pathogens in clinical laboratory, and may improve the management of atypical pneumonia.

A recombinase polymerase amplification-based assay for rapid detection of Chlamydia psittaci.

Chlamydia psittaci is a zoonotic agent of systemic wasting disease in birds and atypical pneumonia in mammalians including humans, constituting a public health risk. A rapid diagnostic assay would be beneficial in screening C. psittaci in the field. In this study, we developed a probe-based recombinase polymerase amplification (RPA) assay for the rapid detection of C. psittaci. The specific primer pairs and probe targeting the conserved region of the outer membrane protein A gene were designed and applied to the real-time real-time RPA assay. The test can be performed at 39°C for 20 min using a portable device, with sensitivities approaching 100 copies of DNA molecules per reaction, with no cross-reaction with other pathogens. The clinical performance of the RPA assay was evaluated in an outbreak of C. psittaci and has high accuracy levels in field applications. The epidemic C. psittaci strains were classed into 2 genotypes: A and C. Collectively, this study offers a promising approach in screening for C. psittaci both in a laboratory setting and in field settings, and RPA can be used as an effective clinical test to monitor outbreaks in domestic fowl populations.

Fatal Chlamydia psittaci infection in a domestic kitten.

Chlamydia psittaci has not been reported to cause disease in domestic cats, to our knowledge. In contrast, C. felis infection is common in domestic cats and typically results in conjunctivitis, upper respiratory tract infection, and less frequently pneumonia. Herein, we report the pathologic findings and diagnostic features of a fatal case of psittacosis in a 7-wk-old domestic kitten. The animal was 1 of a litter of 5 that, together with the queen, were yielded to a pet rescue center in Wyoming. Over a period of ~3 wk, the kittens and queen became sick, thin, and icteric prior to death, despite antimicrobial treatments. Postmortem evaluation of a kitten revealed necrosuppurative hepatitis with Gimenez stain-positive intracellular bacteria, nonsuppurative pneumonia, and mild leptomeningitis. The diagnosis of psittacosis was made by 16S rRNA PCR using multiple primer sets and sequencing from liver. Psittacosis should be considered a differential diagnosis in domestic cats with intracellular bacterial hepatitis and interstitial pneumonia.

A 25-year retrospective study of Chlamydia psittaci in association with equine reproductive loss in Australia.

Introduction. Chlamydia psittaci is primarily a pathogen of birds but can also cause disease in other species. Equine reproductive loss caused by C. psittaci has recently been identified in Australia where cases of human disease were also reported in individuals exposed to foetal membranes from an ill neonatal foal in New South Wales.Hypothesis/Gap Statement. The prevalence of C. psittaci in association with equine reproductive over time and in different regions of Australia is not known.Aim. This study was conducted to detect C. psittaci in equine abortion cases in Australia using archived samples spanning 25 years.Methodology. We tested for C. psittaci in 600 equine abortion cases reported in Australia between 1994 to 2019 using a Chlamydiaceae real-time quantitative PCR assay targeting the 16S rRNA gene followed by high-resolution melt curve analysis. Genotyping and phylogenetic analysis was performed on positive samples.Results. The overall prevalence of C. psittaci in material from equine abortion cases was 6.5 %. C. psittaci-positive cases were detected in most years that were represented in this study and occurred in Victoria (prevalence of 7.6 %), New South Wales (prevalence of 3.9 %) and South Australia (prevalence of 15.4 %). Genotyping and phylogenetic analysis showed that the C. psittaci detected in the equine abortion cases clustered with the parrot-associated 6BC clade (genotype A/ST24), indicating that infection of horses may be due to spillover from native Australian parrots.Conclusion. This work suggests that C. psittaci has been a significant agent of equine abortion in Australia for several decades and underscores the importance of taking appropriate protective measures to avoid infection when handling equine aborted material.

Chlamydiosis in a Gouldian Finch (Erythrura gouldiae).

Avian chlamydiosis is an infection caused by obligate intracellular and Gram-negative bacteria belonging to the family Chlamydiaceae and has been reported in more than 450 avian species distributed in 30 orders. In particular, a high prevalence of infection has been demonstrated in wild passerine populations, including both asymptomatic and clinically ill individuals, suggesting a role of these avian species as important carriers. In May 2018, avian chlamydiosis was diagnosed in a 1-year-old male Gouldian finch (Erythrura gouldiae) at the Turlock Branch of the California Animal Health and Food Safety Laboratory System. The bird belonged to an outdoor aviary with mixed avian species, including Gouldian finches, doves (Geopelia cuneata and Spilopelia chinensis), and psittacines (Aratinga, Psittacula, Pyrrhura, and Trichoglossus sp.). Severe respiratory distress and mortality were noted among the finches. Gross and histopathologic lesions were concentrated in the liver and spleen, with a mild involvement of the upper respiratory tract. Chlamydia spp. were detected in the spleen and kidney by real-time PCR and were further confirmed by immunohistochemistry. Subsequently, Chlamydia psittaci was isolated from the liver and spleen and characterized as a CP3-like strain (genotype B). In addition, viral particles compatible with circovirus were identified in the liver by direct electron microscopy. To the authors' knowledge, this is the first report of avian chlamydiosis with hepatic viral particles consistent with circovirus infection in a Gouldian finch.

Reporte de caso- Clamidiosis en un pinzón diamante de Gould (Chloebia gouldiae). La clamidiosis aviar es una infección causada por bacterias intracelulares y Gramnegativas obligadas que pertenecen a la familia Chlamydiaceae y se ha reportado en más de 450 especies de aves distribuidas en 30 órdenes. En particular, se ha demostrado una alta prevalencia de infección en poblaciones de paseriformes silvestres, incluyendo individuos asintomáticos y clínicamente enfermos, lo que sugiere un papel de estas especies aviares como portadores importantes. En mayo del año 2018, se diagnosticó clamidiosis aviar en un pinzón diamante de Gould (Chloebia gouldiae) de un año de edad remitido a la sede en Turlock del Sistema de Laboratorios de Salud Animal y Seguridad Alimentaria del Estado de California. El ave pertenecía a un aviario al aire libre con especies mixtas de aves, incluyendo los diamantes de Gould, palomas (Geopelia cuneata y Spilopelia chinensis) y psitacinas (Aratinga, Psittacula, Pyrrhura y Trichoglossus sp.). Se observaron problemas respiratorios severos y mortalidad entre los pinzones. Las lesiones macroscópicas e histopatológicas se concentraron en el hígado y el bazo, con problemas leves en el tracto respiratorio superior. Se detectó Chlamydia spp. en el bazo por PCR en tiempo real y fueron confirmados por inmunohistoquímica. Posteriormente, se aisló Chlamydia psittaci del hígado y el bazo y se caracterizó como una cepa de tipo CP3 (genotipo B). Además, se identificaron partículas virales compatibles con circovirus en el hígado mediante microscopía electrónica directa. Según el conocimiento de los autores, este es el primer informe de clamidiosis aviar con partículas virales hepáticas compatibles con infección por circovirus en un pinzón diamante de Gould.

Human psittacosis in Japan: notification trends and differences in infection source and age distribution by gender, 2007 to 2016.

PURPOSE: Psittacosis is a bacterial zoonosis caused by Chlamydia (Chlamydophila) psittaci that infects birds. Although potentially fatal, infections can be reduced by controlling the source of infection. We therefore described the epidemiology of psittacosis, focusing on the infection source. METHODS: We descriptively analyzed psittacosis cases reported through national surveillance in Japan from 2007 to 2016. We also analyzed Chlamydia psittaci prevalence among captive psittaciformes during the same period. RESULTS: One hundred eleven cases were reported, and the annual number and notification rate of psittacosis declined. While 58% were male and the median age was 61 years, the median age differed by gender (males: 63 years, females: 53 years), with more female cases in those aged <50 years. In addition, the most common infection source differed by gender (men: columbiformes; women: psittaciformes). The decline in notifications was associated with a decline in psittaciformes-associated cases, with a concomitant decline in female cases. The prevalence of C. psittaci among captive psittaciformes also decreased over the period. CONCLUSIONS: We found important differences in the epidemiology of psittacosis by gender, and the recent decrease in notifications correlated with decreasing C. psittaci prevalence in birds. Risk communications for psittacosis should consider the current epidemiology regarding gender, age, and infection source.

Chlamydia psittaci in fulmars on the Faroe Islands: a causative link to South American psittacines eight decades after a severe epidemic.

A psittacosis epidemic linked to fulmar hunting occurred on the Faroe Islands in the 1930s. This study investigates a plausible explanation to the 20% human mortality in this outbreak. Phylogenetic analysis showed that Chlamydia psittaci isolated from fulmars were closely related to the highly virulent 6BC strains from psittacines and are compatible with an acquisition by fulmars of an ancestor of the 6BC clade in the 1930s. This supports the hypothesis that the outbreak on the Faroe Islands started after naïve fulmars acquired C. psittaci from infected dead parrots thrown overboard when shipped to Europe in the 1930s.

The application of metagenomic next-generation sequencing in diagnosing Chlamydia psittaci pneumonia: a report of five cases.

BACKGROUND: Chlamydia psittaci pneumonia is a zoonotic infectious disease caused by Chlamydia psittaci. Diagnostic tools, including culture, serologic test and PCR-based methods, are available but prone to false negative results. CASE PRESENTATION: This report included five cases of Chlamydia psittaci pneumonia. Symptoms and signs common to all 5 cases included fever, coughing, generalized muscle ache, and most notably, inflammatory infiltration of the lungs upon chest CT and X-ray. Metagenomic next-generation sequencing (mNGS) revealed the presence of Chlamydia psittaci in biopsy lung tissue in 3 cases and bronchoalveolar lavage fluid in the remaining 2 cases. Three patients responded to doxycycline plus moxifloxacin; two patients responded to moxifloxacin alone. CONCLUSIONS: mNGS could be used to diagnose Chlamydia psittaci pneumonia.

Recent history of psittacosis in Australia: expanding our understanding of the epidemiology of this important globally distributed zoonotic disease.

Psittacosis is a human systemic disease caused by infection with Chlamydia psittaci. Shortly after reports emerged of a global pandemic associated with contact with imported parrots, Australian researchers including Macfarlane Burnet and others demonstrated that C. psittaci was widespread in Australian parrots. Australian cases over the last two decades have revealed that environmental exposure and contact with infected horses are also risk factors in an increasingly complicated epidemiological picture for this zoonotic disease.

Gestational psittacosis: A case report and literature review.

Gestational psittacosis is a rare disease that is associated with significant maternal and fetal morbidity and mortality. Currently, there is no examination method which allows for a quick diagnosis. We report a case of gestational psittacosis that could not be diagnosed as psittacosis during treatment and resulted in maternal and fetal death despite intensive treatment. We also reviewed 23 cases of gestational psittacosis. Fetal and maternal mortality was 82.6% (19/23) and 8.7% (2/23), respectively. In pregnant women with high fever and flu-like symptoms, we should suspect Chlamydia psittaci infection if at least one of the following is present; contact with sheep, parrots, parakeets or goats; normal or moderately decreased leucocyte count, thrombocytopenia and hepatic and/or renal dysfunction; cough and/or lobe consolidation or infiltration on chest X-ray. Antibiotic therapy with macrolide prenatally, macrolide or tetracycline postnatally and termination of pregnancy should be considered.

Detection of Chlamydia psittaci and Chlamydia ibidis in the Endangered Crested Ibis (Nipponia nippon).

Chlamydia spp. are a group of obligate intracellular pathogens causing a number of diseases in animals and humans. Avian chlamydiosis (AC), caused by Chlamydia psittaci (C. psittaci) as well as new emerging C. avium, C. gallinacea and C. ibidis, have been described in nearly 500 avian species worldwidely. The Crested Ibis (Nipponia nippon) is a world endangered avian species with limited population and vulnerable for various infections. To get a better understanding of the prevalence of Chlamydia spp. in the endangered Crested Ibis, faecal samples were collected and analysed. The results confirmed that 20.20% (20/99) of the faecal samples were positive for Chlamydiaceae and were identified as C. ibidis with co-existence of C. psittaci in one of the 20 positive samples. In addition, ompA sequence of C. psittaci obtained in this study was classified into the provisional genotype Matt116, while that of C. ibidis showed high genetic diversity, sharing only 77% identity with C. ibidis reference strain 10-1398/6. We report for the first time the presence of C. ibidis and C. psittaci in the Crested Ibis, which may indicate a potential threat to the endangered birds and should be aware of the future protection practice.