458
Brief communications
of papovavirus infection can often be made on the basis of clinical signs; postmortem findings, including light and electron microscopic lesions; and serology. Acknowledgement. I thank Dr. R. B. Davis, Department of Avian Medicine, College of Veterinary Medicine, University of Georgia, Athens, for doing the serologic tests.
References 1. Bemier G, Morin M, Marsolais G: 1981, A generalized inclusion body disease in the budgerigar (Melopsittacus undulatus) caused by a papovavirus-like agent. Avian Dis 251083-1092. 2. Bemier G, Morin M, Marsolais G: 1984, Papovavirus induced feather abnormalities and skin lesions in the budgerigar: clinical and pathological findings. Can Vet J 25307-310. 3. Bozeman LH, Davis RB, Gaudry D, et al.: 1981, Characterization of a papovavirus isolated from fledgling budgerigars. Avian Dis 25:972-980. 4. Davis RB: 1983, Budgerigar fledgling disease (BFD). Proc 32nd West Poult Dis Conf, p. 104. 5. Davis, RB, Bozeman LH, Gaudry D, et al.: 1981, A viral disease of fledgling budgerigars. Avian Dis 25: 179-183. 6. Dykstra MJ, Bozeman LH: 1982, A light and electron microscopic examination of budgerigar fledgling disease virus in tissue and cell culture. Avian Path01 11: 11-28. 7. Forshaw D, Wylie SL, Pass DA: 1988, Infection with a virus resembling papovavirus in Gouldian finches (Erythrura gouldiae). Aust Vet J 65:26-28. 8. Gaskin JM: 1989, Psittacine viral diseases: a perspective. Zoo and Wildlife Med 20:249-264. 9. Gough JF: 1989, Outbreaks of budgerigar fledgling disease in three aviaries in Ontario. Can Vet J 30:672-674. 10. Graham DL, Calnek BW: 1987, Papovavirus infection in handfed parrots: virus isolation and pathology. Avian Dis 31:398410. 11. Hirai K, Nonaka H, Fukushi H, et al.: 1984, Isolation of a
papovavirus-like agent from young budgerigars with feather abnormalities. Jpn J Vet Sci 46:577-582. 12. Jacobson ER, Hines SA, Quesenberry K, et al.: 1984, Epomitic of papova-like virus-associated disease in a psittacine nursery. J Am Vet Med Assoc 185:1337-1341. 13. Johnston KM, Riddell C: 1986, Intranuclear inclusion bodies in finches. Can Vet J 27:432-434. 14. Krautwald ME, Kaleta EF: 1985, Relationship(s) of French moult and early virus induced mortality in nestling budgerigars. Proc 8th Int Congr World Vet Poult Assoc Jerusalem, p. 115. 15. Lafeber TJ: 1965, Bird clinic: the budgerigar. Anim Hosp 1: 122-132. 16. Lehn H, Miiller H: 1986, Cloning and characterization of budgerigar fledgling disease virus, an avian polyomavirus. Virology 151:362-370. 17. McDowell EM, Trump BF: 1976, Histologic fixatives suitable for diagnostic light and electron microscopy. Arch Pathol Lab Med 100:405-414. 18. Niagro FD, Ritchie BW, Latimer KS, et al.: 1990, Polymerase chain reaction detection of PBFD virus and BFD virus in suspect birds. Proc Assoc Avian Vet, pp. 25-37. 19. Pascucci S, Maestrini N, Misciattelli ME, Giovannetti L: 1983, Malattia da virus papova-simile nel pappagallino ondulato (Melopsittacus undulatus). Clin Vet (Milan) 106:38-41. 20. Pass DA: 1985, A papova-like virus infection of lovebirds (Agapornis sp.). Aust Vet J 62:3 18-3 19. 21. Pass DA, Prus SE, Riddell C: 1987, A papova-like virus infection of splendid parakeets (Neophema splendida). Avian Dis 31:680-684. 22. Randall CJ, Lees S, Inglis DM: 1987, Papovavirus-like infection in budgerigars (Melopsittacus undulatus). Avian Pathol 16: 623-633. 23. Sztojko V, Saghy E, Meder M, et al.: 1985, A hullamos papagaj (Melopsittacus undulatus) papovavirus okozta megbetegedesenek hazai megallapitasa. Magy Allatorv Lapja 40:59-63. 24. Wainwright PO, Lukert PD, Davis RB, Villegas P: 1987, Serological evaluation of some Psittaciformes for budgerigar fledgling disease virus. Avian Dis 31:673-676.
J Vet Diagn Invest 4:458-460 (1992)
A comparison of isolation and a commercial ELISA for the diagnosis of chlamydiosis in psittacine birds Thomas R. Haven, Kenneth W. Mills, Elizabeth S. Williams, Michael P. Driscoll, Richard S. Kingston The problem of identifying birds that are persistently or latently infected with Chlamydia and the desire for more rapid diagnosis of diseased animals has resulted in the development of many new test procedures.2,4,5,7,12,14 As advances in technology supply diagnostic laboratories with an increasFrom the Wyoming State Veterinary Laboratory, Department of Veterinary Sciences, University of Wyoming, 1174 Snowy Range Rd., Laramie, WY 82070 (Haven, Mills, Williams), Broadmoore East Veterinary Clinic, 4218 E. Pershing Blvd., Cheyenne, WY 82070 (Driscoll), and the Department of Animal and Nutritional Sciences, Veterinary Diagnostic Laboratory, Kendall Hall, University of New Hampshire, Durham, NH 03824 (Kingston). Received for publication January 30, 1992.
ing variety of tests, each laboratory must choose tests appropriate to their circumstances. Each method attempts to maximize sensitivity and specificity while minimizing cost, time, specialized equipment, and technical expertise required to perform the test. Several recent comparisons of the available techniques for diagnosing Chlamydial infections have not made the choice easy. A commercial enzyme-linked immunosorbent assay (ELISA)a was reported to be as sensitive as culture in diagnosing Chlamydia psittaci infections in birds submitted for necropsy.5 The same test kit was reported to be “ideally suited for office use” in cases of human conjunctivitis; although it was “not quite as sensitive as culture,” it was less costly, faster, and easier to perform.3 Culture or peroxidase-antiperoxidase (PAP) examination of tissues were
Downloaded from vdi.sagepub.com at NEW YORK UNIV LIBRARY on April 27, 2015
Brief communications Table 1.
459
Comparison of ELISA with isolation procedures at 3 laboratories* to confirm Chlamydial infections in psittacine birds.
reported as equally sensitive, and both were more sensitive than latex agglutination or PAP on cloacal swab specimens. Direct immunofluorescence of fecal smears had a relative sensitivity of only 62.5% as compared with cell culture.14 In late July 1989, an aviary in Wyoming with a continuing problem of C. psittaci infections requested testing to determine if any asymptomatic birds were shedding Chlamydia. This opportunity was used to compare the efficacy of the routine Chlamydial isolation protocol in use at the Wyoming State Veterinary Laboratory (WSVL) to that of a commercial ELISA. The 31 birds remaining on the premises consisted of parrots (10), lovebirds (6), cockatiels (5), conures (5), cockatoos (3), and budgerigars (2). Duplicate oral and cloacal swabsb were collected from each bird. One set of swabs was processed for isolation by placing the material collected into 1.5 ml of Bovarnick’s medium. 1 The second set was left in the protective sleeves. All samples were transported on ice to WSVL. Chlamydia isolation was performed according to accepted procedures2,4,8,10-l3 within 24 hours of specimen collection. At WSVL, 24-well platesc with 12-mm cover glassesd inserted into each well were used as the incubation chamber. Centrifugation at 2,000 x g for 60 minutes and the inclusion of the metabolic inhibitor colchicinee at 2 µg/ml in the maintenance medium were used to enhance the infection rate. McCoy cellsf were stained at 4 days postinfection by direct fluorescent antibody staining.g,9 The dry swabs were received on ice at the Veterinary Diagnostic Laboratory at the University of New Hampshire (NHVDL) within 48 hours of collection and frozen at -70 C until processed for ELISA testing. Processing for ELISA followed package instructions. Of the 62 samples, 5 (8%) were isolation positive from 3 different birds (2 lovebirds, 1 cockatiel); 2 birds were positive on both the oral and cloacal swabs, and 1 lovebird tested positive only from the oral swab. None tested positive by ELISA. Subsequently, 10 tissue culture supernatants were submitted to the National Veterinary Services Laboratory (NVSL) for Chlamydia isolation and to NHVDL for ELISA testing. The supernatants submitted were 4 positive and 6 negative (at WSVL) Chlamydia isolation cases from the WSVGNHVDL study. Two additional Chlamydia-positive super-
natants were sent to NHVDL for ELISA testing. These were isolates from birds that had died soon after being purchased from the aviary under study. Tissues from both animals were direct fluorescent antibody and Chlamydia isolation positive, and both had microscopic lesions of airsaculitis and chronic hepatitis consistent with a diagnosis of Chlamydia infection. The owner of 1 of these birds was diagnosed by his physician as having psittacosis. Four of the 10 samples from asymptomatic birds were isolation positive for Chlamydia at NVSL. Three of these were also isolation positive at WSVL. The positive case from WSVL that was negative at NVSL was an oral swab whose corresponding cloacal swab was positive at both institutions. However, 1 sample that was positive at NVSL was culture negative at WSVL. Chlamydia were isolated from 7 of the 12 tissue culture supernatants (58%) tested at WSVL and/or NVSL. The 2 samples in disagreement were both ELISA negative at NHVDL. Two of the 12 supernatants (16%) tested ELISA positive. One was positive at all 3 test sites; the other sample was not tested at NVSL but was culture positive at WSVL. The culture-positive sample associated with the owner that had psittacosis tested negative by ELISA. An overall agreement of 88% (Table 1) between Chlamydia isolation and ELISA was observed; but this figure may be misleading because of the low number of positive birds in the study. Isolation was clearly superior to ELISA in the diagnosis of Chlamydial infection in live birds. Because the ELISA utilized tests for the presence of antigen already present in a sample, whereas isolation will amplify the amount of antigen, isolation should be a more sensitive test, as was verified by this study. The relatively poor performance of the ELISA may be related to the type of swab used in this study. Different types of swabs, storage temperatures, and processing delay times affect isolation results6 and may also affect ELISA results. Both laboratories performing isolations were in agreement only 80% of the time, emphasizing that isolation is not a perfect test. ELISA has many advantages, compared with isolation, that make it a promising test for the future; however, attempting isolation for Chlamydia is justified by the number of cases in which a diagnosis was achieved by isolation in the absence of ELISA confirmation.
Downloaded from vdi.sagepub.com at NEW YORK UNIV LIBRARY on April 27, 2015
Brief communications
460
Acknowledgement. We thank the Diagnostic Virology Laboratory at the National Veterinary Services Laboratory, Ames, IA, for assistance provided.
Sources and manufacturers a. Kodak Surecello Chlamydia Test Kit, Clinical Products Division, Eastman Kodak, Rochester, NY. b. S/P Culturette, American Scientific Products, Division of Travenol Laboratories, McGraw Park, IL. c. Falcon 3047 Tissue culture plates, Curtin Matheson Scientific, Aurora, CO. d. Bellco Glass, Vineland, NJ. e. GIBCO Laboratories, Life Technologies, Grand Island, NY. f. CRL 1696 McCoy cells: American Type Culture Collection, Rockville, MD. g. National Veterinary Services Laboratory, Ames, IA.
References 1. Bovarnick MR, Miller JC, Snyder JC: 1950, The influence of certain salts, amino acids, sugars, and proteins on the stability of rickettsiae. J Bacterial 59:509-522. 2. Brown S, White J: 1982, Diagnosis of avian psittacosis using cell culture technique. Proc Annu Meet Am Assoc Vet Lab Diagn 25:151-158. 3. Hammerschlag MR, Gelling M, Roblin PM, et al.: 1990, Comparison of Kodak Surecell Chlamydia test kit with culture for the diagnosis of Chlamydial conjunctivitis in infants. J Clin Microbiol 28:1441-1442. 4. Kennedy GA: 1985, Laboratory diagnosis of Chlamydial diseases. Proc Annu Meet Am Assoc Vet Lab Diagn 28:421-436.
5. Kingston RS: 1989, Rapid detection of Chlamydia in pet birds and cats using the Kodak Surecell Chlamydia test kit. Proc Annu Meet Am Assoc Vet Lab Diagn 32:12. 6. Mahony JB, Chemesky MA: 1985, Effect of swab type and storage temperature on the isolation of Chlamydia trachomatis from clinical specimens. J Clin Microbiol 22:865-867. 7. Moore FM, McMillan MC, Petrak ML: 1991, Comparison of culture, peroxidase-antiperoxidase reaction, and latex agglutination methods for the diagnosis of chlamydiosis in pet birds. J Am Vet Med Assoc 199:71-73. 8. Moulder JW: 1991, Interaction of Chlamydiae and host cells in vitro. Microbiol Rev 55: 143-190. 9. Riggs JL: 1979, Immunofluorescent staining. In: Diagnostic procedures for viral, ricketsial, and Chlamydial infections, ed. Lennette EH, Schmidt NJ, 5th ed., pp. 141-170. American Public Health Association, Washington, DC. 10. Schachter JS: 1989, Reports from the symposium on avian chlamydiosis. J Am Vet Med Assoc 195: 1501-1576. 11. Schacter J, Dawson CR: 1979, Psittacosis-lymphogranuloma venerum agents. In: Diagnostic procedures for viral, ricketsial, and Chlamydial infections, ed. Lennette EH, Schmidt NJ, 5th ed., pp. 1021-1059. American Public Health Association, Washington, DC. 12. Spears P, Storz J: 1979, Chlamydia psittaci: growth characteristics and enumeration of serotypes 1 and 2 in cultured cells. J Infect Dis 140:959-967. 13. Storz J: 1971, Chlamydia and Chlamydial-induced diseases, 1st ed., pp. 79-88. Charles C Thomas, Springfield, IL. 14. Woods WW, Dotson JF, Castro AE: 1989, A rapid monoclonal immunofluorescence assay for Chlamydia psittaci in fecal smears from psittacine birds. J Vet Diagn Invest 1:150-153.
J Vet Diagn Invest 4:460-463 (1992)
Isolation of Chlamydia psittaci from pleural effusion in a dog Francisco Arizmendi, James E. Grimes, Roberta L. Relford Chlamydia psittaci is an obligate intracellular parasite that is responsible for producing various signs in domestic and wild mammals and birds. The organism can infect the respiratory, urogenital, and intestinal tracts and the central nervous system and is also responsible for feline conjuncti3,7,10,15,17,19 vitis and ovine and bovine polyarthritis. Diagnosis of C. psittaci infection is based on isolation of the organism, antigen detection and serologic testing, or preferably a combination of both. Isolation is accomplished by inoculation of chicken embryos, mice, guinea pigs, and cell cultures. 1,9,14,17 Serologic tests, such as complement fixation (CF) and microimmunoflourescence (IFA), and antigen detection tests, such as immunoperoxidase techniques and enzyme-linked immunosorbent assay (ELISA) are used to diagnose chlamydial infection in different species.2,5,7,8,11-13,16,18 From the Texas Veterinary Medical Diagnostic Laboratory, PO Drawer 3040, College Station, TX 77841-3040 (Arizmendi, Grimes), and the Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843 (Relford). Received for publication January 31, 1992.
In a natural Chlamydial infection in England,4 C. psittaci was isolated from feces of an adult dog that had eaten the carcasses of dead budgerigars from an aviary in which birds were diagnosed as positive for C. psittaci. That dog developed a CF titer of 128. In Germany, a Chlamydial agent was determined to be the cause of conjuctivitis in 1 dog,10 and antibodies against C. psittaci were detected by an ELISA in 20% of 1,127 serum samples from dogs. 18 In a seroepidemiological survey of C. psittaci in dogs and cats-in Japan, 34 of 359 dogs had CF titers ranging from 16 to 64.5 Chlamydiosis, as a naturally occurring infectious disease, has not been reported in the canine population of the United States. However, experimental evidence suggests that dogs are susceptible to C. psittaci. Clinical signs were induced in dogs inoculated parenterally with a Chlamydial agent of ovine polyarthritis. 11 The agent was reisolated from brain, liver, spleen, kidney, and lung tissue, the intestinal tract, and synovial fluid. The CF titer rose from < 4 to 256 at 9 days postinoculation. The antibody titer decreased to 16 by 1 year after the initial inoculation. Clinical responses, lesions, and frequency of Chlamydia reisolation from dogs inoculated with an ovine Chlamydia strain have also been studied.20 Focal
Downloaded from vdi.sagepub.com at NEW YORK UNIV LIBRARY on April 27, 2015
Brief communications
of papovavirus infection can often be made on the basis of clinical signs; postmortem findings, including light and electron microscopic lesions; and serology. Acknowledgement. I thank Dr. R. B. Davis, Department of Avian Medicine, College of Veterinary Medicine, University of Georgia, Athens, for doing the serologic tests.
References 1. Bemier G, Morin M, Marsolais G: 1981, A generalized inclusion body disease in the budgerigar (Melopsittacus undulatus) caused by a papovavirus-like agent. Avian Dis 251083-1092. 2. Bemier G, Morin M, Marsolais G: 1984, Papovavirus induced feather abnormalities and skin lesions in the budgerigar: clinical and pathological findings. Can Vet J 25307-310. 3. Bozeman LH, Davis RB, Gaudry D, et al.: 1981, Characterization of a papovavirus isolated from fledgling budgerigars. Avian Dis 25:972-980. 4. Davis RB: 1983, Budgerigar fledgling disease (BFD). Proc 32nd West Poult Dis Conf, p. 104. 5. Davis, RB, Bozeman LH, Gaudry D, et al.: 1981, A viral disease of fledgling budgerigars. Avian Dis 25: 179-183. 6. Dykstra MJ, Bozeman LH: 1982, A light and electron microscopic examination of budgerigar fledgling disease virus in tissue and cell culture. Avian Path01 11: 11-28. 7. Forshaw D, Wylie SL, Pass DA: 1988, Infection with a virus resembling papovavirus in Gouldian finches (Erythrura gouldiae). Aust Vet J 65:26-28. 8. Gaskin JM: 1989, Psittacine viral diseases: a perspective. Zoo and Wildlife Med 20:249-264. 9. Gough JF: 1989, Outbreaks of budgerigar fledgling disease in three aviaries in Ontario. Can Vet J 30:672-674. 10. Graham DL, Calnek BW: 1987, Papovavirus infection in handfed parrots: virus isolation and pathology. Avian Dis 31:398410. 11. Hirai K, Nonaka H, Fukushi H, et al.: 1984, Isolation of a
papovavirus-like agent from young budgerigars with feather abnormalities. Jpn J Vet Sci 46:577-582. 12. Jacobson ER, Hines SA, Quesenberry K, et al.: 1984, Epomitic of papova-like virus-associated disease in a psittacine nursery. J Am Vet Med Assoc 185:1337-1341. 13. Johnston KM, Riddell C: 1986, Intranuclear inclusion bodies in finches. Can Vet J 27:432-434. 14. Krautwald ME, Kaleta EF: 1985, Relationship(s) of French moult and early virus induced mortality in nestling budgerigars. Proc 8th Int Congr World Vet Poult Assoc Jerusalem, p. 115. 15. Lafeber TJ: 1965, Bird clinic: the budgerigar. Anim Hosp 1: 122-132. 16. Lehn H, Miiller H: 1986, Cloning and characterization of budgerigar fledgling disease virus, an avian polyomavirus. Virology 151:362-370. 17. McDowell EM, Trump BF: 1976, Histologic fixatives suitable for diagnostic light and electron microscopy. Arch Pathol Lab Med 100:405-414. 18. Niagro FD, Ritchie BW, Latimer KS, et al.: 1990, Polymerase chain reaction detection of PBFD virus and BFD virus in suspect birds. Proc Assoc Avian Vet, pp. 25-37. 19. Pascucci S, Maestrini N, Misciattelli ME, Giovannetti L: 1983, Malattia da virus papova-simile nel pappagallino ondulato (Melopsittacus undulatus). Clin Vet (Milan) 106:38-41. 20. Pass DA: 1985, A papova-like virus infection of lovebirds (Agapornis sp.). Aust Vet J 62:3 18-3 19. 21. Pass DA, Prus SE, Riddell C: 1987, A papova-like virus infection of splendid parakeets (Neophema splendida). Avian Dis 31:680-684. 22. Randall CJ, Lees S, Inglis DM: 1987, Papovavirus-like infection in budgerigars (Melopsittacus undulatus). Avian Pathol 16: 623-633. 23. Sztojko V, Saghy E, Meder M, et al.: 1985, A hullamos papagaj (Melopsittacus undulatus) papovavirus okozta megbetegedesenek hazai megallapitasa. Magy Allatorv Lapja 40:59-63. 24. Wainwright PO, Lukert PD, Davis RB, Villegas P: 1987, Serological evaluation of some Psittaciformes for budgerigar fledgling disease virus. Avian Dis 31:673-676.
J Vet Diagn Invest 4:458-460 (1992)
A comparison of isolation and a commercial ELISA for the diagnosis of chlamydiosis in psittacine birds Thomas R. Haven, Kenneth W. Mills, Elizabeth S. Williams, Michael P. Driscoll, Richard S. Kingston The problem of identifying birds that are persistently or latently infected with Chlamydia and the desire for more rapid diagnosis of diseased animals has resulted in the development of many new test procedures.2,4,5,7,12,14 As advances in technology supply diagnostic laboratories with an increasFrom the Wyoming State Veterinary Laboratory, Department of Veterinary Sciences, University of Wyoming, 1174 Snowy Range Rd., Laramie, WY 82070 (Haven, Mills, Williams), Broadmoore East Veterinary Clinic, 4218 E. Pershing Blvd., Cheyenne, WY 82070 (Driscoll), and the Department of Animal and Nutritional Sciences, Veterinary Diagnostic Laboratory, Kendall Hall, University of New Hampshire, Durham, NH 03824 (Kingston). Received for publication January 30, 1992.
ing variety of tests, each laboratory must choose tests appropriate to their circumstances. Each method attempts to maximize sensitivity and specificity while minimizing cost, time, specialized equipment, and technical expertise required to perform the test. Several recent comparisons of the available techniques for diagnosing Chlamydial infections have not made the choice easy. A commercial enzyme-linked immunosorbent assay (ELISA)a was reported to be as sensitive as culture in diagnosing Chlamydia psittaci infections in birds submitted for necropsy.5 The same test kit was reported to be “ideally suited for office use” in cases of human conjunctivitis; although it was “not quite as sensitive as culture,” it was less costly, faster, and easier to perform.3 Culture or peroxidase-antiperoxidase (PAP) examination of tissues were
Downloaded from vdi.sagepub.com at NEW YORK UNIV LIBRARY on April 27, 2015
Brief communications Table 1.
459
Comparison of ELISA with isolation procedures at 3 laboratories* to confirm Chlamydial infections in psittacine birds.
reported as equally sensitive, and both were more sensitive than latex agglutination or PAP on cloacal swab specimens. Direct immunofluorescence of fecal smears had a relative sensitivity of only 62.5% as compared with cell culture.14 In late July 1989, an aviary in Wyoming with a continuing problem of C. psittaci infections requested testing to determine if any asymptomatic birds were shedding Chlamydia. This opportunity was used to compare the efficacy of the routine Chlamydial isolation protocol in use at the Wyoming State Veterinary Laboratory (WSVL) to that of a commercial ELISA. The 31 birds remaining on the premises consisted of parrots (10), lovebirds (6), cockatiels (5), conures (5), cockatoos (3), and budgerigars (2). Duplicate oral and cloacal swabsb were collected from each bird. One set of swabs was processed for isolation by placing the material collected into 1.5 ml of Bovarnick’s medium. 1 The second set was left in the protective sleeves. All samples were transported on ice to WSVL. Chlamydia isolation was performed according to accepted procedures2,4,8,10-l3 within 24 hours of specimen collection. At WSVL, 24-well platesc with 12-mm cover glassesd inserted into each well were used as the incubation chamber. Centrifugation at 2,000 x g for 60 minutes and the inclusion of the metabolic inhibitor colchicinee at 2 µg/ml in the maintenance medium were used to enhance the infection rate. McCoy cellsf were stained at 4 days postinfection by direct fluorescent antibody staining.g,9 The dry swabs were received on ice at the Veterinary Diagnostic Laboratory at the University of New Hampshire (NHVDL) within 48 hours of collection and frozen at -70 C until processed for ELISA testing. Processing for ELISA followed package instructions. Of the 62 samples, 5 (8%) were isolation positive from 3 different birds (2 lovebirds, 1 cockatiel); 2 birds were positive on both the oral and cloacal swabs, and 1 lovebird tested positive only from the oral swab. None tested positive by ELISA. Subsequently, 10 tissue culture supernatants were submitted to the National Veterinary Services Laboratory (NVSL) for Chlamydia isolation and to NHVDL for ELISA testing. The supernatants submitted were 4 positive and 6 negative (at WSVL) Chlamydia isolation cases from the WSVGNHVDL study. Two additional Chlamydia-positive super-
natants were sent to NHVDL for ELISA testing. These were isolates from birds that had died soon after being purchased from the aviary under study. Tissues from both animals were direct fluorescent antibody and Chlamydia isolation positive, and both had microscopic lesions of airsaculitis and chronic hepatitis consistent with a diagnosis of Chlamydia infection. The owner of 1 of these birds was diagnosed by his physician as having psittacosis. Four of the 10 samples from asymptomatic birds were isolation positive for Chlamydia at NVSL. Three of these were also isolation positive at WSVL. The positive case from WSVL that was negative at NVSL was an oral swab whose corresponding cloacal swab was positive at both institutions. However, 1 sample that was positive at NVSL was culture negative at WSVL. Chlamydia were isolated from 7 of the 12 tissue culture supernatants (58%) tested at WSVL and/or NVSL. The 2 samples in disagreement were both ELISA negative at NHVDL. Two of the 12 supernatants (16%) tested ELISA positive. One was positive at all 3 test sites; the other sample was not tested at NVSL but was culture positive at WSVL. The culture-positive sample associated with the owner that had psittacosis tested negative by ELISA. An overall agreement of 88% (Table 1) between Chlamydia isolation and ELISA was observed; but this figure may be misleading because of the low number of positive birds in the study. Isolation was clearly superior to ELISA in the diagnosis of Chlamydial infection in live birds. Because the ELISA utilized tests for the presence of antigen already present in a sample, whereas isolation will amplify the amount of antigen, isolation should be a more sensitive test, as was verified by this study. The relatively poor performance of the ELISA may be related to the type of swab used in this study. Different types of swabs, storage temperatures, and processing delay times affect isolation results6 and may also affect ELISA results. Both laboratories performing isolations were in agreement only 80% of the time, emphasizing that isolation is not a perfect test. ELISA has many advantages, compared with isolation, that make it a promising test for the future; however, attempting isolation for Chlamydia is justified by the number of cases in which a diagnosis was achieved by isolation in the absence of ELISA confirmation.
Downloaded from vdi.sagepub.com at NEW YORK UNIV LIBRARY on April 27, 2015
Brief communications
460
Acknowledgement. We thank the Diagnostic Virology Laboratory at the National Veterinary Services Laboratory, Ames, IA, for assistance provided.
Sources and manufacturers a. Kodak Surecello Chlamydia Test Kit, Clinical Products Division, Eastman Kodak, Rochester, NY. b. S/P Culturette, American Scientific Products, Division of Travenol Laboratories, McGraw Park, IL. c. Falcon 3047 Tissue culture plates, Curtin Matheson Scientific, Aurora, CO. d. Bellco Glass, Vineland, NJ. e. GIBCO Laboratories, Life Technologies, Grand Island, NY. f. CRL 1696 McCoy cells: American Type Culture Collection, Rockville, MD. g. National Veterinary Services Laboratory, Ames, IA.
References 1. Bovarnick MR, Miller JC, Snyder JC: 1950, The influence of certain salts, amino acids, sugars, and proteins on the stability of rickettsiae. J Bacterial 59:509-522. 2. Brown S, White J: 1982, Diagnosis of avian psittacosis using cell culture technique. Proc Annu Meet Am Assoc Vet Lab Diagn 25:151-158. 3. Hammerschlag MR, Gelling M, Roblin PM, et al.: 1990, Comparison of Kodak Surecell Chlamydia test kit with culture for the diagnosis of Chlamydial conjunctivitis in infants. J Clin Microbiol 28:1441-1442. 4. Kennedy GA: 1985, Laboratory diagnosis of Chlamydial diseases. Proc Annu Meet Am Assoc Vet Lab Diagn 28:421-436.
5. Kingston RS: 1989, Rapid detection of Chlamydia in pet birds and cats using the Kodak Surecell Chlamydia test kit. Proc Annu Meet Am Assoc Vet Lab Diagn 32:12. 6. Mahony JB, Chemesky MA: 1985, Effect of swab type and storage temperature on the isolation of Chlamydia trachomatis from clinical specimens. J Clin Microbiol 22:865-867. 7. Moore FM, McMillan MC, Petrak ML: 1991, Comparison of culture, peroxidase-antiperoxidase reaction, and latex agglutination methods for the diagnosis of chlamydiosis in pet birds. J Am Vet Med Assoc 199:71-73. 8. Moulder JW: 1991, Interaction of Chlamydiae and host cells in vitro. Microbiol Rev 55: 143-190. 9. Riggs JL: 1979, Immunofluorescent staining. In: Diagnostic procedures for viral, ricketsial, and Chlamydial infections, ed. Lennette EH, Schmidt NJ, 5th ed., pp. 141-170. American Public Health Association, Washington, DC. 10. Schachter JS: 1989, Reports from the symposium on avian chlamydiosis. J Am Vet Med Assoc 195: 1501-1576. 11. Schacter J, Dawson CR: 1979, Psittacosis-lymphogranuloma venerum agents. In: Diagnostic procedures for viral, ricketsial, and Chlamydial infections, ed. Lennette EH, Schmidt NJ, 5th ed., pp. 1021-1059. American Public Health Association, Washington, DC. 12. Spears P, Storz J: 1979, Chlamydia psittaci: growth characteristics and enumeration of serotypes 1 and 2 in cultured cells. J Infect Dis 140:959-967. 13. Storz J: 1971, Chlamydia and Chlamydial-induced diseases, 1st ed., pp. 79-88. Charles C Thomas, Springfield, IL. 14. Woods WW, Dotson JF, Castro AE: 1989, A rapid monoclonal immunofluorescence assay for Chlamydia psittaci in fecal smears from psittacine birds. J Vet Diagn Invest 1:150-153.
J Vet Diagn Invest 4:460-463 (1992)
Isolation of Chlamydia psittaci from pleural effusion in a dog Francisco Arizmendi, James E. Grimes, Roberta L. Relford Chlamydia psittaci is an obligate intracellular parasite that is responsible for producing various signs in domestic and wild mammals and birds. The organism can infect the respiratory, urogenital, and intestinal tracts and the central nervous system and is also responsible for feline conjuncti3,7,10,15,17,19 vitis and ovine and bovine polyarthritis. Diagnosis of C. psittaci infection is based on isolation of the organism, antigen detection and serologic testing, or preferably a combination of both. Isolation is accomplished by inoculation of chicken embryos, mice, guinea pigs, and cell cultures. 1,9,14,17 Serologic tests, such as complement fixation (CF) and microimmunoflourescence (IFA), and antigen detection tests, such as immunoperoxidase techniques and enzyme-linked immunosorbent assay (ELISA) are used to diagnose chlamydial infection in different species.2,5,7,8,11-13,16,18 From the Texas Veterinary Medical Diagnostic Laboratory, PO Drawer 3040, College Station, TX 77841-3040 (Arizmendi, Grimes), and the Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843 (Relford). Received for publication January 31, 1992.
In a natural Chlamydial infection in England,4 C. psittaci was isolated from feces of an adult dog that had eaten the carcasses of dead budgerigars from an aviary in which birds were diagnosed as positive for C. psittaci. That dog developed a CF titer of 128. In Germany, a Chlamydial agent was determined to be the cause of conjuctivitis in 1 dog,10 and antibodies against C. psittaci were detected by an ELISA in 20% of 1,127 serum samples from dogs. 18 In a seroepidemiological survey of C. psittaci in dogs and cats-in Japan, 34 of 359 dogs had CF titers ranging from 16 to 64.5 Chlamydiosis, as a naturally occurring infectious disease, has not been reported in the canine population of the United States. However, experimental evidence suggests that dogs are susceptible to C. psittaci. Clinical signs were induced in dogs inoculated parenterally with a Chlamydial agent of ovine polyarthritis. 11 The agent was reisolated from brain, liver, spleen, kidney, and lung tissue, the intestinal tract, and synovial fluid. The CF titer rose from < 4 to 256 at 9 days postinoculation. The antibody titer decreased to 16 by 1 year after the initial inoculation. Clinical responses, lesions, and frequency of Chlamydia reisolation from dogs inoculated with an ovine Chlamydia strain have also been studied.20 Focal
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