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Detection of Chlamydia psittaci (ovis) antigen in tissue sections and McCoy cells using streptavidin-biotin and the IMAGEN staining method.
Two procedures for identifying chlamydial antigen in infected McCoy cells and tissue sections are described. Both the streptavidin-biotin method and the IMAGEN test clearly detected chlamydial antigen in ovine placental tissue sections and in infected McCoy cells before they could be detected by conventional stains. The streptavidin-biotin method is lengthy, but specific and sensitive, and the slides can be kept indefinitely for later examination. By contrast, the IMAGEN test is a single step procedure which requires fluorescent microscopy and slides can only be kept for up to 1 week before examination.
Chlamydia psittaci: prevailing problems in pathogenesis.
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Studies on the early phase of the pathogenesis of ovine enzootic abortion in the non-pregnant ewe.
The initial phase of infection of non-pregnant sheep by Chlamydia psittaci (ovis) was studied by inoculating naïve and previously exposed sheep by the oro-nasal and subcutaneous routes with the BS isolate of C. psittaci (ovis). Naive animals exhibited a marginal rise in temperature and seroconversion was detected by enzyme-linked immunosorbent assay (ELISA) as early as 9 days after inoculation. Chlamydaemia was detected using culture and an antigen detection ELISA. No faecal shedding of chlamydiae was detected in ewes kept up to 3 weeks after infection. Although chlamydial antigen was demonstrated in epithelial cells or lymphocytes in lungs, liver, spleen, kidney, abomasum, jejunum, tonsils and suprapharyngeal, mandibular, parotid and mesenteric lymph nodes of some of the naïve sheep using an ELISA and streptavidin-biotin and immunofluorescent staining techniques, the organism could not be cultured from these tissues. No chlamydial antigen was demonstrated in any of the tissues of the previously exposed sheep nor in uninoculated controls. It is concluded that previously infected sheep are capable of completely eliminating subsequent infection and that chlamydiae localize in a variety of tissues of infected naïve sheep, especially within epithelial cells and lymphocytes.
Epizootic bovine abortion in a dairy herd: characterization of a Chlamydia psittaci isolate and antibody response.
A chlamydial agent was recovered from the placental cotyledons of an aborting cow from a 100-cow dairy herd in Cumbria. Immunoblotting analysis of purified elementary bodies of the isolate revealed a reactivity pattern typical of serotype I Chlamydia psittaci strains. Nucleotide sequencing of the major outer membrane protein (MOMP) gene further confirmed the isolate, BA1, as a serotype I strain. The sequence was identical to that of the type strain of ovine enzootic abortion, B577. In both the antigenic and MOMP sequencing analyses BA1 was distinguishable from serotype II C. pecorum strains. A sequential series of sera obtained from the aborting cow, from which BA1 was recovered, was analysed by immunoblotting against the homologous isolate, and demonstrated reactivity to major chlamydial antigens over a 110-day period. Close contact between ruminant species on the farm suggested that the C. psittaci strain may have been transmitted to cattle from infected sheep.
Recombinant ovine interferon gamma inhibits the multiplication of Chlamydia psittaci in ovine cells.
The local production of interferon-gamma (IFN-gamma) in sheep in response to Chlamydia psittaci was measured by cannulation of the efferent lymph duct draining the site of challenge inoculation. Peak production of IFN-gamma (256 U/ml) was detected 24 h after challenge. Based on these physiological data, functional studies were carried out in vitro to determine the effect of recombinant ovine (rOv) IFN-gamma on the multiplication of C. psittaci in ovine fibroblasts. IFN-gamma inhibited the multiplication of C. psittaci in ovine cells over a range of concentrations (250 U/ml to 2.5 U/ml) in a dose-dependent manner. The inhibition of chlamydial multiplication was most pronounced when cells were treated with rOvIFN-gamma for 24 h before infection. The addition of exogenous L-tryptophan (500 micrograms/ml) to cultures within 48th of infection abrogated the anti-chlamydial effect of rOvIFN-gamma thus suggesting that tryptophan deprivation is an anti-chlamydial mechanism induced by rOvIFN-gamma in these ovine cells.
Immune responses of the ovine lymph node to Chlamydia psittaci. A cellular study of popliteal efferent lymph.
The popliteal efferent lymphatics were cannulated in sheep of two categories, seronegative or immune to Chlamydia psittaci. Following subcutaneous injection of live C. psittaci or control material into the draining area of the popliteal node, sequential samples of efferent lymph were collected and analysed. Both categories of sheep responded to C. psittaci with increased outputs of lymphocytes and blast cells. Numbers of blast cells rose both absolutely and as a proportion of the total. Plasmablasts increased in number only in seronegative sheep. Outputs of total T cells (CD5+), helper T cells (CD4+), cytotoxic/suppressor T cells (CD8+) and non-helper, non-suppressor T cells (T19) were maximal 4 and 7 days after challenge in immune and seronegative sheep, respectively. Proportionally, CD4+ T cells declined, CD8+ T cells increased and T19 cells were unaltered with time after infection. Chlamydial antigens could not be demonstrated in the cells of efferent lymph by an immunoperoxidase method. The results of this preliminary study show that both T and B cell responses are involved in immunity to C. psittaci.
Pathogenesis of Chlamydia psittaci infection in sheep: detection of the organism in a serial study of the lymph node.
Sixteen sheep were inoculated subcutaneously over the left prefemoral lymph node with an abortifacient strain of Chlamydia psittaci. Groups of four animals were killed after 3, 6, 12 and 18 days. Four of eight sheep which received a control inoculum were killed on day 6 and four on day 18. The left and right prefemoral lymph nodes were removed and weighed and portions taken from each for examination by the polymerase chain reaction (PCR), by culture, and by histopathological and immunohistochemical methods. The left prefemoral lymph nodes enlarged after the injection of C. psittaci, with the group mean weight on day 6 being the greatest and that on day 18 being normal. Examination by "nested" PCR showed samples from these nodes to be positive, except for one animal killed on day 3 and one on day 12. Live organisms, however, were not cultured from any of the samples collected. C. psittaci antigen was detected immunohistochemically in three of four nodes on day 3, in each of four on day 6, and in two of four on both days 12 and 18. Nodes from the contralateral side remained normal, as did those from unchallenged control sheep, and no antigen or DNA was detected in them.
Interferon-gamma mediates long-term persistent Chlamydia psittaci infection in vitro.
The long-term anti-chlamydial effects of recombinant ovine interferon gamma (rOvIFN-gamma) were studied in ovine ST-6 fibroblasts infected with the S26/3 strain of Chlamydia psittaci. Chlamydial multiplication was assessed by enzyme-linked immunosorbent assay analysis of supernate lipopolysaccharide, titration of inclusion-forming units in culture supernates, and enumeration of inclusion bodies in cultured cells at 7-day intervals. Concentrations of 250 and 1000 U/ml of rOvIFN-gamma resulted in a microbistatic inhibition of C. psittaci growth, which appeared to become microbicidal when rOvIFN-gamma was maintained in the cultures for 14 days or more. There were no signs of C. psittaci multiplication when cultures were maintained in 25 or 100 U/ml of rOvIFN-gamma. However, subsequent removal of rOvIFN-gamma from these cultures resulted in a re-emergence of viable, infectious chlamydiae, which eventually killed all the fibroblasts. This re-emergence was more rapid in cultures initially treated with 25 U/ml of rOvIFN-gamma than in those treated with 100 U/ml.
Infective endocarditis. A review of selected topics.
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Respiratory chlamydia: the management of an outbreak.
This investigation was undertaken in response to the occurrence of nine cases of respiratory chlamydial infection in 8 months within the district of Dudley. All nine cases of respiratory chlamydia were due to Chlamydia psittaci, not Chlamydia pneumoniae. Five cases had avian exposure, but no other aetiological factors were identified. Faecal specimens were obtained from only two of the implicated birds and were negative. Two local aviaries were identified as sources of implicated birds. The two aviaries were themselves linked. Bird faecal specimens were taken from the two implicated aviaries and were both positive for Chlamydia psittaci. Appropriate public health control measures were introduced in these aviaries. A press statement was released to identify and advise, by telephone, those who purchased birds from the aviaries.
SOME DISEASES COMMON TO ANIMALS AND MAN.
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Outbreak of chlamydia infection in rural Australian town.
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Ornithosis; isolation of the virus from a case.
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Ornithosis in two railway guards, an occupational hazard.
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