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Chlamydial serum IgG, IgA and local IgA antibodies in patients with genital-tract infections measured by solid-phase radioimmunoassay.

A solid-phase radioimmunoassay (RIA) for IgG and IgA class antibodies to Chlamydia trachomatis was developed with C. trachomatis serotype L2 as antigen. The assay was sensitive, reproducible and correlated well with an immunofluorescence test (r = 0.85). Serum IgG antibodies were detected in 79% of Chlamydia isolation-positive versus 43% of isolation-negative male patients with urethritis, and and serum IgA antibodies in 53% and 21%, respectively. Urethral IgA antibodies, measured from specimens taken for chlamydial isolation, could be detected in 94% and 38%, respectively. From 737 male urethral and 909 female cervical secretions screened for the presence of IgA antibodies, about half were isolation and IgA negative. Only 4% (6/151) of male and 5.4% (2/37) of female isolation-positive specimens were IgA negative. The determination of local IgA antibodies may be used as a screening test in chlamydial genital infections.

Antibodies, Bacterial↗

Seroepidemiology of feline chlamydiosis by microimmunofluorescence assay with multiple strains as antigens.

The prevalence of anti-chlamydia antibodies was examined in 232 cat sera collected in 1985 and from 1993 to 1995 from laboratories and veterinary hospitals located in 11 prefectures of Japan. The antibodies were determined by an indirect microimmunofluorescence test using six strains of feline Chlamydia: one strain each of avian- and guinea pig-derived C. psittaci and one strain each of C.pecorum, C.pneumoniae and C.trachomatis. Positive rates of IgG antibodies to chlamydiae were 34.4% in 1985 and 16.5-21.4% from 1993 to 1995. Positive rates of IgM antibodies to chlamydiae were 8.2% in 1985 and 6.6-14.3% from 1993 to 1995. Variations in antibody reactivity to the different feline strains were observed. The results suggest the wide prevalence of chlamydial infection in cats in Japan, and antigenic diversity in the feline strains of C.psittaci.

Animals↗

Polymerase chain reaction for the detection of Chlamydia psittaci in the feces of budgerigars.

The polymerase chain reaction (PCR) targeting the ompA gene of Chlamydia psittaci was evaluated for its ability to detect chlamydiae in fecal specimens of budgerigars as compared with isolation procedures using cell culture and embryonated egg inoculations. Several procedures for PCR template DNA preparation were compared so as to determine their detection levels for chlamydiae propagated in cell culture in the presence of fecal materials. Tween-20 and proteinase K treatments followed by centrifugation of the template DNA were found to be an appropriate procedure for DNA preparation for primary PCR. Subsequent nested PCR was shown to detect 4.8 IFU/ml or 84 particles/ml of chlamydiae. Chlamydiae in 50 fecal specimens from apparently healthy budgerigars were examined by nested PCR and several other methods. Nested PCR detected chlamydiae at a higher rate (12/50, 24%) than the isolation procedure in embryonated eggs (6/50, 12%). Primary PCR combined with the isolation procedure in cell culture gave a detection rate (5/50, 10%) similar to that of isolation from embryonated eggs. Detection rates by primary PCR (1/50, 2%) and in cell culture (0%) were inferior to the other procedures.

Animals↗

Experimental Chlamydia psittaci infection of Japanese quail.

Japanese quail were used for the infection model of avian chlamydiosis. One-day-old Japanese quail were highly susceptible to lethal infection by a Chlamydia psittaci strain of budgerigar origin upon inoculation via the air sac route with 10(4.1) FFU of the organism, showing an acute and lethal course with chlamydial propagation. In contrast, 7-day-old quail developed resistance to the infection as shown by the lack of lethal effect with the same dose. The resistance of 7-day-old birds was abolished by immunosuppressive treatment with cyclophosphamide. Upon inoculation with a sublethal dose of 10(2.1) FFU, latent infection was established in 1-day-old birds with a minimum number of the organism. The latent infection in the birds was converted to the lethal form by treatment with cyclophosphamide along with chlamydial propagation and suppression of antibody production.

Aging↗

Seroepidemiological investigation of feline chlamydiosis in cats and humans in Japan.

The prevalence of chlamydia antibodies in Japan was investigated in 215 cat sera, consisting of 88 sera of stray cats and 127 sera of pet cats, and 2,184 human sera, taken from 2,003 general persons and 181 small animal clinic veterinarians, by microimmunofluorescence (MIF) testing with Chlamydia psittaci Fe/Pn1 of feline origin and Prk/6BC of avian origin as antigens. The prevalence rates of anti-Fe/Pn1 antibodies were 45.5% in stray cats, 17.3% in pet cats, 1.7% in general persons and 8.8% in small animal clinic veterinarians. The prevalence rates of anti-Prk/6BC antibodies were 51.1% in stray cats, 15.0% in pet cats, 3.1% in general persons and 5.0% in small animal clinic veterinarians. These results suggested that feline chlamydia infection is widely spread in cats especially in stray cats in Japan, and suggested that feline chlamydiosis could be transmitted to people who are in close contact with infected cats.

Animals↗

Genetic diversity and epizootiology of Chlamydophila psittaci prevalent among the captive and feral avian species based on VD2 region of ompA gene.

To study genetic diversity and occurrence of Chlamydophila psittaci, a total of 1,147 samples from 11 avian orders including 53 genera and 113 species of feral and captive birds were examined using ompA gene based nested PCR. Three types of chlamydiae: C. psittaci (94.12%), C. abortus (4.41%) and unknown Chlamydophila sp. (1.47%) were identified among 68 (5.93%) positive samples (Psittaciformes-59, Ciconiiformes-8 and Passeriformes-1). Based on nucleotide sequence variations in the VD2 region of ompA gene, all 64 detected C. psittaci strains were grouped into 4 genetic clusters. Clusters I, II, III and IV were detected from 57.35%, 19.12%, 10.29% and 7.35% samples respectively. A single strain of unknown Chlamydophila sp. was found phylogenetically intermediate between Chlamydophila species infecting avian and mammalian hosts. Among Psittaciformes, 28 out of 81 tested species including 10 species previously unreported were found to be chlamydiae positive. Chlamydiosis was detected among 8.97% sick and 48.39% dead birds as well 4.43% clinically normal birds. Therefore, it was observed that though various genetically diverse chlamydiae may cause avian chlamydiosis, only a few C. psittaci strains are highly prevalent and frequently associated with clinical/subclinical infections.

Amino Acid Sequence↗

Seroprevalence of Chlamydia psittaci-specific antibodies in small stock in Namibia--epidemiological study with an enzyme-linked immunosorbent assay (ELISA).

An IgG (H+L)-ELISA was applied as a screening test for antibodies against Chlamydia (C.) psittaci in sera of goats and sheep in Namibia. In 576 (27.3%) of a total of 2,107 sera (299 = 25.2% of 1,185 caprine and 277 = 30.0% of 922 ovine sera) chlamydial antibodies could be detected. 86% of all farms tested revealed seropositive animals. Chlamydial infections were prevalent in all the geographical regions tested. The infection rates per State Veterinary District varied from 12.0% (Otjiwarongo) to 50.0% (Otavi) in goats and from 13.3% (Otjiwarongo) to 41.7% (Windhoek) in sheep. The regional distribution of chlamydial infections was not related to geographical or climatic factors. Sera from herds showing symptoms indicative for chlamydial infections showed significantly higher antibody rates (35% in goats and 39% in sheep) than sera from herds without health problems (18% in goats and 24% in sheep). Considering only sera from farms with clinical history of chlamydiosis, high seroprevalences were correlated to the symptoms abortion and keratoconjunctivitis. As in other countries, enzootic abortion seems to be the main manifestation of chlamydial infection in small ruminants in Namibia. C. psittaci might also play a considerable role in the etiology of infectious keratoconjunctivitis, whereas association with other clinical entities seems to be rare.

Animals↗

Diagnosis of avian chlamydiosis: specificity of the modified Giménez staining on smears and comparison of the sensitivity of isolation in eggs and three different cell cultures.

For the diagnosis of chlamydiosis in dead and live birds different methods were compared for their sensitivity and specificity. The specificity of the modified Giménez staining and the direct immunofluorescence (DIF) test for direct demonstration of Chlamydia psittaci in organ, cloacal and/or conjunctival smears was examined. The sensitivity of the isolation of Chlamydia psittaci in 6 days embryonated specific pathogen free (SPF) chicken eggs, Buffalo Green Monkey (BGM) cell line, McCoy cell line and Vero cell line was compared. On smears, the direct immunofluorescence test was more specific than the modified Giménez staining. The concordance between the results of both detection methods was 80%. The BGM cell culture was the most sensitive artificial host for isolation of Chlamydia psittaci, followed by the embryonated eggs, the Vero cell line and the McCoy cell line. The concordance between the results of isolation in BGM cell culture and eggs was 96.5%, while it was 86% between the results of isolation in BGM cell culture and Vero cell culture and only 65.5% between the results of isolation in BGM cell culture and McCoy cell culture. For dead bird species, chlamydiosis could be diagnosed more often using DIF on smears than with isolation. The concordance between the results of the DIF on smears and isolation followed by DIF was 91%.

Animals↗

Comparison of different serological methods for the determination of antibodies to Chlamydia psittaci in pigeon sera.

Several methods for detecting antibodies to Chlamydia psittaci using sera from pigeons were compared with regard to their sensitivity, specificity and efficacy. The reference method used was indirect immunofluorescence (IFI), which uses as antigen intracellular inclusions in McCoy cell monolayers. Indirect micro-immunofluorescence (MIF) using as antigen C. psittaci cultured on yolk sacs of embryonated eggs, direct and indirect complement fixation tests (CFT and ICFT, respectively), and enzyme-linked immunosorbent assay (ELISA) were compared to the reference method. Indirect micro-immunofluorescence proved to be the most efficient method, while ELISA was the most sensitive, though showing a very low specificity. No statistically significant difference was found in comparing the two complement fixation tests. The use of ICFT to check for the negative results obtained in CFT is questioned. Possible reasons for the different results with the methods used are discussed.

Animals↗

Endometritis in cattle experimentally induced by Chlamydia psittaci.

On the day of estrus, eight virgin heifers received intrauterine inoculations of yolk sac propagated Chlamydia psittaci strain BovEnd 11/88 isolated from the uterus of a slaughter cow. All heifers developed purulent vaginal discharge which persisted for 3 to 7 weeks. Chlamydiae or chlamydial antigen were detected in vaginal and uterine discharges of infected animals by culture or Capture ELISA, while other bacterial pathogens were not found. In sera of the chlamydia-infected heifers marked increases in antibody titres against the chlamydial genus-specific LPS-antigen were found by ELISA and complement fixation test. Six heifers were artificially inseminated in 5 successive cycles beginning at the first estrus following intrauterine inoculation. In two of the infected heifers spontaneous healing of endometritis occurred after 5 estrus cycles. Only these animals conceived after the 5th breeding, whereas in the remaining four animals a chlamydia-associated chronic endometritis was recognized as the cause of infertility in the 19th and 26th week p.i. at slaughter. Two control heifers which remained clinically normal after intrauterine exposure to sterile yolk sac-suspensions conceived at the 1st and 2nd service, respectively.

Animals↗

Polymerase chain reaction (PCR) detection of porcine Chlamydia trachomatis and ruminant Chlamydia psittaci serovar 1 DNA in formalin-fixed intestinal specimens from swine.

In previous studies chlamydiae were detected immunohistologically in the gut of 66 out of 311 pigs. The aim of the present investigation was the classification of these intestinal porcine chlamydiae. For the study, DNA extracted from 52 paraffin-embedded intestinal tissues was amplified in nested polymerase chain reactions (PCRs) with Chlamydia omp1 genus- and species-specific primers. Some of the amplification products were cloned and sequenced. In 45 cases DNA could be amplified with genus-specific primers. Species-specific PCR and sequencing showed that in 42 cases the chlamydial omp1 genotype was Chlamydia trachomatis. Sequenced DNA fragments were 95-99% identical with the porcine strain S45. In three further cases sequencing analysis provided DNA sequences which were 100% identical with Chlamydia psittaci B577 (serovar 1) omp1 genotype. So far as the authors are aware this is the first report on the occurrence of C. psittaci serovar 1 in pigs.

Animals↗

Relationship between abortions and seroprevalences to selected infectious agents in dairy cows.

The aim of this study was to determine whether the occurrence of abortion is related to the seroprevalence of abortion-causing infectious agents. In a cross-sectional study, cattle from dairy farms in Switzerland that were defined as having an abortion problem were divided into two groups: cows with a history of abortion within the previous 3 months (cases) and cows without a history of abortion (controls). A positive titre to Leptospira spp. was associated with an increased probability of being a case (OR = 1.74, 95% CI = 1.21-2.47). There were interactions between Coxiella burnetti titre and parity, and between Chlamydia psittaci and C. burnetti titre and breed. Multiparous cases after the second lactation with a positive titre to C. burnetti were less likely (OR = 0.42, 95% CI = 0.22-0.82) to be cases. Swiss Browns (Swiss Braunvieh and Brown Swiss) with a positive titre to C. psittaci and Swiss Browns with a positive titre to C. burnetti were more likely (OR = 1.63, 95% CI = 1.13-2.37 and OR = 1.79, 95% CI = 1.15-2.78, respectively) to be cases. Parity alone was not associated with the occurrence of abortion.

Abortion, Veterinary↗