Experimental enteric infection of gnotobiotic piglets with a Chlamydia psittaci strain of avian origin

The pathogenicity of a Chlamydia psittaci isolate of pigeon origin was assessed using a litter of gnotobiotic piglets. At 3 days of age, six piglets were inoculated intragastrically with egg-grown chlamydiae, the remaining six pigs were sham-inoculated. The animals were observed for clinical signs, and they were killed and necropsied sequentially between 4 and 15 days of age. Clinical manifestations consisted of slight softening of the faeces between 6 and 10 days post-inoculation (DPI). Immunohistochemistry revealed chlamydial replication predominantly in the small intestine, initially within villous enterocytes, after 4 DPI mostly in the lamina propria. Histopathology showed villous atrophy and increased numbers of inflammatory cells in the gut up to 6 DPI. Chlamydial stages of normal morphology were identified within enterocytes using transmission electron microscopy. An enzyme-linked immunosorbent assay (ELISA) run on faecal samples revealed shedding of chlamydial antigen from 3 until 11 DPI. Systemic dissemination of Chlamydia occurred to a limited extent according to polymerase chain reaction and immunohistochemistry results of several extraintestinal organs. Corresponding histopathological changes were minimal. Sera of all pigs were negative for anti-chlamydial antibodies using a complement fixation test. In conclusion, inoculation of this isolate in gnotobiotic piglets resulted in a productive enteric infection with mild lesions, weak systemic dissemination, and faecal shedding, indicating the pig as a potential host for avian chlamydiae.     

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。     

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.     

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。     

Use of ovotransferrin as an antimicrobial in turkeys naturally infected with Chlamydia psittaci, avian metapneumovirus and Ornithobacterium rhinotracheale

Respiratory pathogens are difficult to control in large-scale turkey production. This report describes a clinical trial of antimicrobial ovoTF aerosol on a large Belgian turkey farm. ovoTF was administered to reduce Chlamydia psittaci (C. psittaci) infections and to study the impact of this action on the occurrence of Ornithobacterium rhinotracheale (O. rhinotracheale) and avian metapneumovirus (aMPV) infections. Two subsequent broods were included; (i) a control brood receiving no ovoTF and (ii) an ovoTF brood receiving ovoTF aerosol (5mg/animal) at the age of 2 weeks, continuing daily for 12 days. Twenty-four one-day-old toms of the control and ovoTF brood were tagged and monitored for 15 weeks. The control brood experienced two periods of respiratory disease, the first (2-3 weeks of age) due to C. psittaci and the second (8-17 weeks of age) in the presence of C. psittaci, O. rhinotracheale and maybe aMPV. Extensive antibiotic treatment was needed in 2, 8 and 9 week-old toms. In the ovoTF brood, toms stayed healthy until the age of 9 weeks, whereafter respiratory disease occurred in the presence of C. psittaci, O rhinotracheale and aMPV. OvoTF administration: (i) reduced the amount of C. psittaci in the air as demonstrated by bioaerosol monitoring, (ii) prevented respiratory disease during the first half of the brood period, (iii) was associated with 46% reduction of mortality, and (iv) reduced the antibiotic cost. Our results justify additional clinical trials to explore the use of this innovative antimicrobial strategy for poultry.     

Characterization of DNA fragment from Chlamydia psittaci avian strain which shows high homology with hypB gene of Chlamydia.

A study was performed to characterize DNA fragment No. 17 of C. psittaci strain P-1041 which encoded 42 KD beta-galactosidase fusion protein with type-specific antigenicity. Sequence determination identified a partial open reading frame that spanned about 1,200b. p. nucleotides. Screening the literatures for the nucleotide and deduced amino acid sequences revealed extensive similarity between the DNA fragment of P-1041 and two chlamydial hypB genes. This DNA showed 91.5% homology with C. psittaci GPIC hypB gene in nucleotide sequence and 96.4% homology in deduced amino acid sequence. The hypB gene of C. trachomatis serovar A and the P-1041 DNA fragment showed 81.2% and 91.3% homology in nucleotide and amino acid sequences, respectively. Dot enzyme-linked immunosorbent assay, for the products of deleted DNA fragments defined the coding region for type-specific antigenic polypeptide. In addition, the P-1041 DNA fragment carried a sequence highly homologous (greater than 49%) with other bacterial and plant genes called chaperonin which responds to various stress in cells. From these results, the P-1041 DNA fragment was found to be a part of hypB gene and to encode the region critical for type-specific antigenicity.     

A preliminary survey of Chlamydia psittaci genotypes from native and introduced birds in New Zealand

AIM: To describe the Chlamydia psittaci genotypes in samples from native and introduced birds from New Zealand by analysis of the sequence variation of the ompA gene.

METHODS: DNA was extracted from samples collected from a non-random sample of birds; either swabs from live asymptomatic birds or birds with clinical signs, or formalin-fixed, paraffin-embedded (FFPE) samples from historical post-mortem cases. The presence of C. psittaci in all samples had been confirmed using a quantitative PCR assay. The C. psittaci ompA gene was amplified and sequenced from samples from 26 native and introduced infected birds comprising 12 different species. These sequences were compared to published available C. psittaci genotypes.

RESULTS: Genotypes A and C of C. psittaci were identified in the samples. Genotype A was identified in samples from nine birds, including various native and introduced species. Genotype C was identified in samples from 16 different waterfowl species, and a mixed infection of both genotypes was found in a kaka (Nestor meridionalis). In native birds, C. psittaci infection was confirmed in seven new host species.

CONCLUSIONS AND CLINICAL RELEVANCE: Two genotypes (A and C) of C. psittaci were found in samples from a wider range of both native and introduced species of birds in New Zealand than previously reported. Both genotypes have been globally associated with significant disease in birds and humans. These initial results suggest the host range of C. psittaci in New Zealand birds is under-reported. However, the prevalence of C. psittaci infection in New Zealand, and the associated impact on avian and public health, remains to be determined. There are biosecurity implications associated with the importation of birds to New Zealand if there is a limited diversity of C. psittaci genotypes present.     

Respiratory and pericardial lesions in turkeys infected with avian or mammalian strains of Chlamydia psittaci

Three groups of turkeys were inoculated with strains of C. psittaci (B577, VS1, TT3) from different restriction endonuclease groups. Turkeys were necropsied at 15 times through post-inoculation day 70. Birds infected with the TT3 strain were lethargic and had decreased body weight. After forced exercise, dyspnea was seen in VS1-infected turkeys. Pericarditis was the most severe lesion in TT3-infected birds. Airsacculitis and bronchopneumonia were the most severe lesions in VS1-infected turkeys. Lateral nasal adenitis was in both VS1- and TT3-infected birds. Only mild peribronchial pneumonia was in B577-infected turkeys. Chlamydial antigen, identified by light microscopy using an immunoperoxidase technique, was seen from post-inoculation days 9 through 50 in the lateral nasal gland and at earlier times in other tissue from VS1- and TT3-infected turkeys. No chlamydial antigen was detected in tissue from B577-infected birds. These studies showed that chlamydial strains from different restriction endonuclease groups are associated with distinct disease syndromes in turkeys.     

Direct immunofluorescence tests with counterstains for detection of Chlamydia psittaci in infected avian tissues.

Different methods of preparation and serological evaluation of rabbit globulins for use in fluorescent antibody conjugate and different methods of counterstaining with fluorescent antibody tests were evaluated for detection of Chlamydia psittaci in infected turkey tissues. The agar gel precipitin reaction was that chosen for testing and selecting antiserums to be used for fluorescein isothiocyanate conjugation. The fluorescent antibody staining was most pronounced with conjugate made from globulins precipitated with ammonium sulfate. A direct fluorescent antibody method with Evans blue counterstain correctly identified "coded" specimens of C. psittaci-infected and noninfected turkey air sacs. However, naphthalene black was superior to Evans blue as a counterstain when infected pericardial sacs were tested.     

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.     

Antibody response of the ovine lymph node to experimental infection with an ovine abortion strain of Chlamydia psittaci

After primary infection with Chlamydia psittaci in the draining area of the popliteal lymph node, viable organisms could be isolated from the efferent lymph only before the primary immune response developed. The lymph antibody response, as assayed by the complement fixation and immunofluorescence (IF) antibody tests, showed a rise in titre that peaked approximately 2 weeks after infection. Immunoblotting revealed that antibodies produced during this period were predominantly directed against the major outer membrane protein (MOMP). In secondary infection of convalescent sheep, an elevated IF antibody titre, already present in the lymph and blood, could not be boosted. Viable organisms could not be isolated from these sheep. Antibodies produced reacted to 12-14 bands in the immunoblot profile including the MOMP band. These potentially immunoprotective antigens, particularly MOMP, should be considered as useful candidates for an improved vaccine against ovine enzootic abortion, in further investigations.     

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.     

Characterization of the systemic disease and ocular signs induced by experimental infection with Chlamydia psittaci in cats

In addition to the commonly reported ocular signs, Chlamydia psittaci infection of kittens resulted in fever, lethargy, lameness and reduction in weight gain following ocular instillation of virulent organisms. The appearance of these systemic signs was late with respect to the appearance of ocular symptoms and occurred simultaneously with increasing levels of chlamydia-specific IgG. Measurement of acute phase reactants and IL-6 in plasma indicated that both became elevated concurrent with or slightly after the appearance of fever and remained elevated after the fever began to resolve. Preliminary data also indicated that infectious C. psittaci was present in the blood stream during this time period. The results of ocular instillation of three different levels of C. psittaci (10^3.8, 10^2.8 and 10^1.5 TCID₅₀) indicated that the frequency of infection and the severity of ocular signs were diminished in the group receiving the lowest dose. However, the magnitude of systemic disease was similar in all animals which exhibited clinical signs, irrespective of the dose administered. The immune response to infection included elementary body (EB)-specific lymphocyte proliferation as well as the development of EB-specific IgG and IgM antibodies. The predominant antibody response was to a 45 kDa protein, the major outer membrane protein (MOMP), lipopolysaccharide (LPS), a 58 kDa doublet and 32 and 16–19 kDa proteins.     

Emergency response following suspicion of an avian influenza outbreak

The management of a suspected index case of highly pathogenic avian influenza (AI) is crucial to the subsequent actions aimed at limiting the spread of infection and ultimately in prompt eradication of the virus. In this study, the legislative basis, basic concepts and actions behind a successful disease management programme are reviewed. These include actions at local veterinary headquarters and at farm level which must be coordinated centrally to ensure the flow of information essential to decision making. The success of any emergency intervention strategy is dependent upon the level of preparedness, including action plans to source manpower, equipment and on the degree of communication between relevant parties. Availability of information on the successes and failures in field outbreak management, will inevitably result in improved AI control worldwide.