First detection of Chlamydia psittaci from a wild native passerine bird in New Zealand

AIMS: To undertake disease surveillance for Chlamydia psittaci in native birds as part of a pilot study to examine pathogen diversity on Hauturu-o-Toi/Little Barrier Island. To retrospectively review the Massey University post-mortem database to determine previous cases of avian chlamydiosis in New Zealand.

METHODS:Mistnetting of forest birds was conducted across an elevational gradient on Hauturu-o-Toi/Little Barrier Island. Minitip culture swabs were used to collect cloacal samples from native birds. These swabs were screened for Chlamydia family DNA using two PCR methods. Positive results were sequenced. A retrospective review of the Massey University post-mortem database of all avian cases from 1990 to 2011 was conducted.

RESULTS:Ten native birds including four bellbirds (Anthornis melanura), three rifleman (Acanthisitta chloris), two hihi (Notiomyces cincta), and one whitehead (Mohoua albicilla) were sampled and one otherwise healthy female hihi was positive by both PCR screening methods for Chlamydophila. Sequencing confirmed 99–100% genetic similarity to C. psittaci. A retrospective review of the Massey University post-mortem database revealed no previous diagnoses of avian chlamydiosis in wild native New Zealand birds although it has been detected in captive parrots, and wild and captive exotic pigeons.

CONCLUSIONS:This is the first report of the detection of C. psittaci from a wild native bird in New Zealand. The bird was a Passeriforme from an endangered species that was captured free-living on Little Barrier Island. The incidence of avian chlamydiosis in native birds in New Zealand appears to be very low, based on the retrospective review of the post-mortem database.

CLINICAL RELEVANCE: It is unlikely that avian chlamydiosis is a significant problem for hihi population health. The detection of this organism has greater significance for other more susceptible species on Little Barrier Island and for human health, particularly for conservation workers involved in wildlife translocations. It further suggests that passerine birds may be a reservoir for C. psittaci in New Zealand ecosystems.     

Chlamydophila psittaci infections in humans during an outbreak of psittacosis from poultry in Germany

In 2005, an outbreak of severe respiratory disease in a mixed poultry flock that was infected with Chlamydophila (C.) psittaci led to dissemination of the infection to at least 100 small poultry farms in 11 districts of Central Germany. At the same time, a total of 24 persons in contact with poultry from one of the flocks reported flu-like symptoms to their physician, thus suggesting zoonotic transmission. Within 3 weeks, seven individuals had to be hospitalized, with three of them requiring intensive care. Analysis of ompA sequences from chlamydial isolates and directly from clinical samples revealed the presence of both genotype A and E/B of C. psittaci at the source of the outbreak and in contact flocks. Genotype A was also detected in the three severely ill patients. The findings of the present study demonstrate the high zoonotic potential of avian chlamydiae. To ensure speedy eradication of psittacosis in poultry flocks and effective treatment of infected humans, fast, sensitive and species-specific detection of the causative agent is essential, as well as close collaboration between regional public health services, attending physicians and the diagnostic laboratories involved.     

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.     

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.     

动物鹦鹉热衣原体疫苗研究现状

鹦鹉热衣原体为多种人畜共患病的病原体。衣原体免疫学机制研究表明其主要引发Th1细胞调节的细胞免疫和体液免疫。减毒活疫苗、灭活疫苗的使用在一定程度上控制了其发病率。基因工程疫苗,如重组亚单位苗、核酸疫苗、细茵载体苗等也都在深入研究中。同时,人们也在筛选特异性的抗原,构建各种疫苗载体及选择免疫佐剂和免疫途径等方面做了大量的工作。     

Seroprevalence of Antibodies to Chlamydophila psittaci in Zoo Workers in Brazil

To evaluate the prevalence of antibodies to Chlamydophila psittaci 364 serum samples were collected from veterinarians, biologists, animal scientists, veterinary students, animal keepers and others employees in 20 zoos, and from veterinary practitioners in 10 Brazilian states. Subjects ranged from 15 to 64 years of age, with 268 (74%) males and 96 (26%) females. Chlamydial antibodies were determined by the complement fixation test (CFT) and specific anti‐C. psittaci IgG antibodies were determined by the microimmunoflurescence (MIF) test. Complement fixation test showed 23.9% (87/364) and MIF test showed 4.7% (17/364) positive serum samples. Titres ranged from 16 to 256 in both assays, demonstrating evidence of recent or current infection. Although chlamydial antibodies were detected in workers of seventeen zoos, MIF test only detected specific C. psittaci antibodies in seven of them. Previous psittacosis infection was suspected in eight workers of two zoos, five of whom reported having pneumonia, while employed at the zoos. However, diagnosis was not established in any of these cases in the past. Results indicated the occurrence of infection and previous contact of Brazilian zoo workers with C. psittaci, as well as the zoonotic potential of psittacosis in this risk population. Other studies are necessary to evaluate the risk factors of infection in this population. This seroepidemiological survey confirmed the need to adopt preventive measures to control avian chlamydiosis and protect the health of zoo workers in the country.     

1株鹦鹉热衣原体的冻干、鉴定和分子分型研究

从国家兽医微生物菌(毒)种保藏中心领取1支鹦鹉热衣原体CVCC2410进行冻干和鉴定。将衣原体经卵黄囊接种SPF鸡胚进行复壮和传代,收获卵黄囊膜研磨后分装、冻干,并对冻干衣原体进行了无菌检验、真空度测定、染色特性和形态观察、特异性检验、16S rRNA鉴定、分子分型、衣原体含量测定。结果表明冻干衣原体CVCC2410 2203为C型鹦鹉热衣原体,含量为10^5.50ELD₅₀/0.2mL。本研究完善了该株衣原体的信息,为制定鹦鹉热衣原体的入库标准奠定基础。     

北京市及周边省份家禽鹦鹉热嗜性衣原体流行状况的调查

对北京市周边6省份怀疑感染鹦鹉热嗜性衣原体的鸡鸭血清样品374份、病料81份，分别使用IHA诊断试剂盒、ELISA试剂盒以及抗酸染色试剂和荧光抗体诊断试剂进行了检查，以评价北京市及其周边地区家禽鹦鹉热嗜性衣原体的流行性。结果，上述4种试剂检测出的阳性率依次为24．9％、77．9%、18．5%和38．2％；北京市10份SPF鸡血清的抗体全部为阳性；患病肉鸡、肉鸭气囊样品，蛋鸡输卵管样品的检出率较高。表明，北京市及其周边地区家禽已经感染了鹦鹉热嗜性衣原体，酶联免疫吸附法和荧光抗体染色法能分别提高抗体和抗原的检出率。     

Chlamydia psittaci: a suspected cause of reproductive loss in three Victorian horses

Chlamydia psittaci was detected by PCR in the lung and equine foetal membranes of two aborted equine foetuses and one weak foal from two different studs in Victoria, Australia. The abortions occurred in September 2019 in two mares sharing a paddock northeast of Melbourne. The weak foal was born in October 2019 in a similar geographical region and died soon after birth despite receiving veterinary care. The detection of C. psittaci DNA in the lung and equine foetal membranes of the aborted or weak foals and the absence of any other factors that are commonly associated with abortion or neonatal death suggest that this pathogen may be the cause of the reproductive loss. The detection of C. psittaci in these cases is consistent with the recent detection of C. psittaci in association with equine abortion in New South Wales. These cases in Victoria show that C. psittaci, and the zoonotic risk it poses, should be considered in association with equine reproductive loss in other areas of Australia.     

山东省部分地区家禽鹦鹉热衣原体感染情况调查

为了解山东地区家禽鹦鹉热衣原体(Cps)感染现状,本研究采用间接血凝法(IHA)对2013年~2014年采集自山东潍坊、淄博、济南、临沂、烟台等地区的1 020份鸡、鸭血清样品进行Cps抗体的检测,并对检测数据进行了统计分析;结果显示:IHA测得总阳性率为29.51%(301/1 020);鸡阳性率为25.26%(197/780);鸭阳性率为43.33%(104/240);各地区间阳性率存在一定差异。本调查结果表明,家禽Cps感染在山东地区具有较高的感染率。     

布鲁氏菌和鹦鹉热衣原体双重PCR检测方法的建立与应用

为建立同时检测布鲁氏菌和鹦鹉热衣原体的双重PCR方法,本研究据GenBank上已发表的具有属间特异性的布鲁氏菌bp26基因和鹦鹉热衣原体23S rRNA基因,利用 Primer Premier 5.0软件各设计1对特异性引物,扩增的目的片段长度分别为219和356 bp。通过优化反应条件,建立了能同时检测布鲁氏菌和鹦鹉热衣原体的双重PCR方法。该方法具有较好的特异性和可重复性,对2种基因单重PCR检测敏感性均达到3.1×10²拷贝/反应,双重检测的灵敏度为3.1×10³拷贝/反应。利用该双重PCR方法对流产牛抗凝全血、血清、流产胎儿及奶液共172份临床疑似布鲁氏菌感染的样品进行检测,检测到布鲁氏菌阳性样品53份,鹦鹉热衣原体阳性样品2份,以上这2种病原的阳性检出率分别为30.8%和1.2%,且检测到2种病原混合感染的阳性样品2份,阳性检出率为1.2%。临床应用结果表明,该方法可用来对布鲁氏菌和鹦鹉热衣原体进行同步、快速、灵敏的检测。     

Infection of the Bovine Female Genital Tract with Chlamydia psittaci as a Possible Cause of Infertility

Contents: Chlamydia psittaci was isolated in embryonated chicken eggs via the yolk sac route from ten (16,7%) vaginal andlor endometrial mucosal scrapings of 60 slaughter cows. Simultaneous fecal shedding of chlamydiae was found in four animals. Chlamydial infections of the genital tract were frequent (p < 0,01) when there were endometrial inflammatory lesions together with the failure to detect other bacterial pathogens in the uterus.     

Comparison of the omp I gene of Chlamydia psittaci between isolates in Victorian koalas and other animal species

Objective The objective of this study is to compare the strain of chlamydia causing genital infection in koalas from Victoria with isolates from other animal species.
Design Polymerase chain reaction and restriction enzyme analysis has been used to compare various Chlamydia psittaci isolates from a range of animals and disease syndromes. The isolates used in this study include isolates from three birds, three from aborted sheep, one from polyarthritis, one from bovine abortion, one from feline pneumonitis, three porcine isolates from faeces, polyarthritis and abortion, and three urogenital isolates from Victorian koalas.
Procedure Two polymerase chain reactions were performed, each amplifying a different region of the omp I gene. The first polymerase chain reaction amplified a 144 bp segment of the gene which was then digested with the restriction enzyme Eco R I. The second polymerase chain reaction amplified a larger 1070 bp region of the omp I gene which was digested with two restriction enzymes Alu I and Nde II.
Results and conclusions The results obtained have confirmed that variation in DNA sequence of various animal chlamydia isolates does occur. They have also shown that it is possible to classify isolates, based on their restriction enzyme profiles, into distinct groups.     

Isolation and identification of Chlamydia psittaci from pet birds

Culturing of Chlamydia psittaci from pet birds requires the inoculation of a susceptible living host system with suspensions of various tissues from dead birds or with tracheal and/or cloacal swabs and fresh feces from live birds. Cell cultures have been used as the host system. The most commonly used cell cultures for isolation of C psittaci from pet birds are McCoy and mouse L cells. The sensitivity and specificity of cell culture equals or surpasses embryonating chicken eggs and mice, and results can be obtained in less than 7 days. To obtain satisfactory results, the inoculum must be centrifuged onto the cell cultures at 37 C, and the cells must be treated with a metabolic inhibitor such as colchicine or cycloheximide. Chlamydia psitaci can be detected in infected cells by use of fluorescent antibody, Giemsa, or Gimenez staining.     

Detection of Chlamydophila psittaci from feral pigeons in environmental samples: problems with currently available techniques

Chlamydophila psittaci (Lillie, 1930) Everett et al., 1999, the pathogenic agent of human ornithosis, is widespread in feral pigeon populations and many cases of transmission from feral pigeons to humans have been reported. The aim of the present study was to detect C. psittaci in environmental samples to find out more about possible transmission routes and, therefore, to assess the zoonotic risk for humans. Fecal samples were collected from nest boxes in a feral pigeon loft. Additionally, samples were taken from the feather dust film covering the water surface of public fountains where pigeons regularly bathe. The samples were tested for the presence of chlamydial antigen using an antigen enzyme-linked immunosorbent assay to prove shedding of C. psittaci by feral pigeons. This test detects a genus specific lipopolysaccharide in the outer membrane of the chlamydial bacteria. Samples were tested using the IDEIA PCE Chlamydia Test kit (DakoCytomation) and positive results were verified with IDEIA Chlamydia Blocking Reagents (DakoCytomation). The IDEIA PCE Chlamydia Test yields a high proportion of positive results. However, when IDEIA Chlamydia Blocking was performed, most of the positive results turned out to be negative or could not be interpreted. We conclude that antigen-enzyme-linked immunosorbent assay tests are not suitable for detecting C. psittaci in environmental samples. Previous publications where no blocking test was used should be reconsidered critically.     

A cluster of two psittacosis cases among women farmers exposed to Chlamydia psittaci-infected domestic poultry in Zhejiang Province,China

A cluster of Chlamydia psittaci (C. psittaci) cases was reported in Zhejiang Province, China, 2019. This study evaluates the extent of the outbreak and determines the source of infection. Real-time PCR and sequencing of the ompA gene of C. psittaci were performed to identify the cases, the domesticated poultry and close contacts. The index patient was a 76-year-old woman with chronic vertigo, and Case 2 was a 64-year-old female farmer with herpes zoster. Both women bought psittaci-infected chickens or ducks from the same mobile street vendor and raised them for 10 days and 23 days before fever onset. There were no direct contact between the two women. C. psittaci test was positive for the two patients, one sick chicken, three healthy ducks and the vendor's chicken cage. Phylogenetic analysis showed that all seven C. psittaci positive samples carried identical ompA genotype A of C. psittaci. Of all of the patients' 148 close contacts, none tested positive for C. psittaci, or developed acute respiratory symptoms. Both patients were discharged after a 4-week hospital stay. In conclusion, the source of this cluster was the poultry infected with C. psittaci, which occasionally cause infections in farmers, but inter-human transmission seems unlikely.