Chlamydiosis in Pen-Raised Bobwhite Quail (Colinus Virginianus) and Chukar Partridge (Alectoris Chukar) with High Mortality Author(S): Douglas H

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Chlamydiosis in Pen-Raised Bobwhite Quail (Colinus virginianus) and Chukar Partridge (Alectoris chukar) with High Mortality Author(s): Douglas H. Erbeck and Stacey A. Nunn Source: Avian Diseases, Vol. 43, No. 4 (Oct. - Dec., 1999), pp. 798-803 Published by: American Association of Avian Pathologists, Inc. Stable URL: http://www.jstor.org/stable/1592752 . Accessed: 03/03/2011 11:19

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http://www.jstor.org AVIAN DISEASES43:798-803, 1999

Case Report-

Chlamydiosis in Pen-raised Bobwhite Quail (Colinus virginianus) and Chukar Partridge (Alectoris chukar) with High Mortality DouglasH. Erbeckand StaceyA. Nunn

MurrayState UniversityBreathitt Veterinary Center, 715 North Drive, P.O. Box 2000, Hopkinsville, KY 42241-2000 Received 23 February1999

SUMMARY. In a flock of 12,000 bobwhite quail (Colinusvirginianus) and 7200 chukar partridge (Alectorischukar), the owner had 100% morbidity and 40%-50% mortality in birds between the ages of 2 and 4 wk. Affected birds were stunted and anorexic and had yellow/green diarrhea.Two- and 4-wk-old birds submitted for necropsyall had slight nasal discharge.Histopathologic examinationrevealed mild (bobwhite) to severe (chukar)rhinitis. Immunohistochemistrywas positive for Chlamydiapsittaci in all birds. Chlamydiapsittaci organisms were demonstratedhistopathologically in hematoxylin and eosin and Gimenez- stained slides. Managementsanitation and treatmentwith chlortetracyclinestopped further excessive losses. The owners were also infected. Treatment by their local physician with tetracyclinealleviated symptoms.

RESUMEN. Reportede Caso--Clamidiosis con alta mortalidad en codornices (Colinus virginianus)y en perdices indias (Alectorischukar) criadasen corralesde piso. En una parvadade 12000 codornices blancas (Colinusvirginianus) y 7200 perdicesindias (Alectorischukar), se observ6 una morbilidaddel 100% y una mortalidaddel 40%-50% en aves entre dos y cuatrosemanas de edad. Las aves afectadasmostraban enanismo, inapetencia y tenian diarreade color amarilloverdosa. Las aves de dos a cuatro semanas enviadaspara necropsiapresentaban ligera descarga nasal. El examenhistopatol6gico revel6 una rinitissuave en las codornicesy severaen las perdicesindias. El examen inmunohistoquimicofue positivo para Chlamydiapsittaci en todas las aves. La Chlamydiapsittaci fue observadahistopatol6gi- camente por medio de la tinci6n de Gimenezy de la hematoxilinaeosina. Medidasde manejo sanitario y el tratamiento con clortetraciclinaevitaron mayores perdidas. Los propietarios tambien se infectarony el tratamientocon tetraciclinaalivi6 los sintomas. Key words: Chlamydiapsittaci, chlamydiosisin bobwhite quail, zoonotic chlamydiosis

Chlamydia psittaci is a bacteria that causes and eye discharges, depression, anorexia, respi- disease in birds and humans (1). The organism ratory distress, and yellow diarrhea (7,10). Psit- was originallyisolated from psittacinebirds and tacines, gulls, egrets, and turkeys generally har- the diseasewas called psittacosisor parrotfever bor the more virulent strains (10). Older birds (8). Later, C. psittaci was shown to infect tur- are often nonsymptomaticcarriers and shedders keys and other fowl, and the disease was called (1). Avian strains of C. psittaci can also infect ornithosis (5,6). The organism isolated from humans (1). Pathogenic strains of C. psittaci the different species of birds is currently con- causing clinical signs and mortality have not sidered the same, and the preferred term for the previously been reported in quail (Schwartz, disease in both birds and humans is chlamydio- pers. comm.). sis (9). This case report documents chlamydiosis in The disease is more severe in young birds a flock of bobwhite quail (Colinus virginianus) (10). Clinical signs can include lethargy, nasal and chukar partridge (Alectoris chukar) experi-

798 Chlamydiosisin bobwhiteand chukar 799

encing severe disease with mortality.The own- the birds reported having stuffy head, cough, ers and their adult children who worked with sneezing, mild sore throat, tightness of the the birds were subsequently diagnosed with chest, and headachesof severalweeks' duration. Chlamydiapsittaci infection. A second adult son who worked sporadically with the birds had similar symptoms;however, CASE REPORT his 7-yr-old daughter, who often interacted with the birds, and his pregnantwife, who had History. Four 2-wk-old and four 4-wk-old infrequent contact with the birds, did not dis- bobwhitequail (virginianus)and six 4-wk-old play any symptoms. An adult daughter who chukar partridgewere submittedto Murray never went near the birds, but recalledprevious StateUniversity Breathitt Veterinary Center in contact with a normal-appearingparrot, had June 1998. The birdswere from a game bird the most severe respiratorysigns. When the farmhousing 12,000 bobwhitequail and 7200 owners learnedthat the birds had chlamydiosis, chukar partridgethat suddenly experienced they contacted their local and re- and in physician 100% morbidity 40%-50% mortality quested that the family be tested for the disease. birdsbetween the of 2 and4 wk. The farm ages Serologic results for chlamydia antibodies, im- incubatedand hatchedboth of birds. species munoglobulin G (positive, ?1.10) (LabCorp, The quail were hatchedfrom eggs purchased Louisville, KY), indicated the husband (1.92), from an egg suppliercertified clean for Salmo- his wife (1.33), both adult sons (2.06, 2.13), nella and The chukar Mycoplasma. partridge and adult daughter(3.00) tested positive. Titers were hatched from eggs of 300 breeder birds from the and her 7- maintained in facilities on the farm. pregnant daughter-in-law separate yr-old daughterwere negative (<0.91). Every 2 wk, 1500 quail eggs and 900 partridge in the 2- set Necropsy. Antemortem findings eggs were and incubated. Eggs were incu- wk-old included with bated and the were quail stunting ruffled together, species separated feathers, serous to nasal dis- for serosanguineous hatching. and with swollen Both and chicks were brood- charge, conjunctivitis eyelids partridge quail The 4-wk-old had feath- ed and started in a 32-ft 75-ft (Fig. 1). quail ruffled together by ers with featherloss over the dorsalhead house with six Chicks were brooded spotty partitions. and One chick had at one end of the house where the fresh air body. slight serosanguineous, at the nostrils. One chu- inlets were located. At 2-wk intervals, chicks crusty discharge were moved to the next fartherfrom kar was stunted and two had nasal discharge, partition one of which also had swollen the fresh air At 6 wk, both were eyelids. supply. species Postmortem in the 2-wk old moved to large ground flight pens. Chicks were findings quail included red, mucous fed crushed quail starter pellets, 28% protein, hyperemic conjunctival membranes; sinuses; and that contained virginiamycin(Pfizer, Lee's Sum- congested, hyperemic white urate and fibrinous streaked abdominal mit, MO), 20 g/ton. The first two hatches showed no clinical air sacs. The 4-wk-old quail had congested and nasal serous to signs of infection. During the next two hatches, hyperemic turbinates, clear fibrinous abdominal air and small birds displayed no signs nor symptoms of dis- cloudy sacs, in the femoral muscles. ease until about 2 wk of age. Anorexia,yellow/ streakinghemorrhages two of the 6-wk-old chukar vis- green diarrhea,respiratory discharge, and stunt- Only displayed ible lesions. ing would occur between 2 and 6 wk. Virtually Congested and hyperemicconjunc- 100% of the birds in the brood/starthouse old- tival membranesand nasal turbinateswere ob- er than 2 wk of age displayed clinical signs. served. Moderate numbers of Eimeria sp. oo- Mortality was between 40% and 50%. Birds cysts were present in pooled fecal samples from showing diarrheawere more severely affected. each group of birds. Cryptosporidialorganisms Four or 5 days after transferto flight pens, clin- were not observed. ical signs and mortality abated. There were no Toxicology. Analysis of the crushed starter sequella; those birds that survived would con- feed and pellets fed to the older birds indicated tinue to normal adulthood. that mycotoxinswere within safe limits. By thin Zoonosis. The owners, husband and wife, layer chromatography,aflatoxin B 1, ochratoxin and one adult son who regularlyworked with A, zearalenone, T-2, and Diacetoxyscirpenol 800 D. H. Erbeckand S. A. Nunn

Fig. 1. Two-week-oldbobwhite quail with crusty serosanguineousnasal dischargeand swollen eyelids. were less than 10 ppb and vomitoxin was less ed on precleaned glass slides, and stained with than 1 ppb. hematoxylin and eosin. Microbiology. A genus-specific nested All three groups of birds had histopathologic polymerase chain reaction procedure yielded lesions of the nasal turbinates. One of the 2- negative results for the presence of Mycoplasma wk-old bobwhite quail had slightly hyperplastic genus in lung, trachea, and choanal swabs. turbinate epithelium. In the epithelial tissue Lung mycotic cultures were negative for Asper- were multifocal, mild to heavy accumulations gillus sp. Pseudomonasaeruginosa was isolated of mononuclear leukocytes. Occasional hetero- from lung and nasal swabs of each group and phils were within these areas. In the 4-wk-old from liver from the chukar group. Other path- bobwhite, in two of four nasal turbinates, small ogenic bacteria were not isolated from nasal accumulations of mononuclear leukocytes ap- swabs, trachea, lung, or air sacs. peared in the submucosal lamina. In the 6-wk- Transmission electron microscopy of nasal si- old chukar partridge, the turbinate tissues were nus, trachea, and lung was negative for virus edematous. The epithelial surfaces and sub- particles. Testing methanol-fixed lung impres- mucosal tissues were infiltrated with a fibrinous sion smears for the presence of Chlamydia was exudate containing mixed inflammatory cells. carried out by the fluorescent antibody tech- Small multifocal areas of necrosis with mixed nique. Fluorescein isothiocyanate-conjugated inflammatory cell exudate were present in both monoclonal antibody (Meridian Diagnostics, epithelial and submucosal areas. Inc., Cincinnati, OH) specific for all known Lung tissues were unremarkable in the 2-wk- serotypes of C. psittaci, as well as Chlamydia old quail, whereas one of four lung sections in trachomatis, was used in the procedure. Results the 4-wk-old quail had occasional areas of were weakly positive and reported as equivocal. mononuclear leukocyte infiltration into the Histopathology. Selected tissues, fixed in capillary air spaces. The lungs of the chukar 10% neutral buffered formalin overnight and partridge were unremarkable. Air sac sections processed by conventional methods, were em- from the chukar displayed greatly hyperplastic bedded in paraffin, sectioned at 4 jxm, mount- tissue infiltrated with a fibrinofibrous material. Chlamydiosisin bobwhiteand chukar 801

by the avidin-biotinperoxidase method (Vector Laboratories,Burlingame, CA). Substratewas 3-amino-9-ethyl-carbazole chromagen (Bioge- nex Laboratories, San Ramon, CA). Sections were counterstained lightly with hematoxylin (Ventana Medical Systems, Inc.). Lung macro- phages from each group of birds were positive for C. psittaci. Treatment and results. When lossesfirst occurred,birds were treatedfor 2 wk with ty- losin (Boehringer,St. Joseph,MO), 20 g/ton of feed. Mortalityremained at about50%. Af- ter laboratorydiagnosis, the birdswere treated with chlortetracycline(Durvet, Inc., Blue Springs,MO), 300 g/ton of feed, for chlamydiosis and monensin (Bovatec,Hoffman-La- Roche,Inc., Nutley,NJ), 80 g/ton of feed, for coccidiosis.Thorough cleansing and disinfection of the brooder/grow-outpartitions and water/feedlines with 30% sodium hypochlorite Fig. 2. Lung, capillaryair space macrophagewith (VertexChemical IL) werecon- bodies. Corp., Dupo, multiple Chlamydiae psittaci elementary ducted. Lossesfrom later hatchesdropped to x400. less than 10%. The humanswere treatedwith tetracycline After immunohistochemistrystudies were hydrochloride(Mylan Pharmaceuticals,Inc., positivefor C psittaci,retrospective histopath- Morgantown,WV), 250 mg twicedaily for 15 ologicexamination revealed C. psittaci reticulate days.The husbandand the adultchildren had bodiesand more numerouselementary bodies dramaticreduction of symptomswithin 5 days. (Fig. 2) in a few lung macrophagesin at least The wife reportedonly mild reductionof re- one birdin eachage group and in both species. spiratoryproblems. Two weeks after comple- The Gimenezstain identifiedcharacteristic tion of treatment,the husbandreported return elementarybodies in lung tissues. Chlamydia of respiratorysymptoms. Tetracycline treatment psittaciorganisms stained bright red and back- was resumedand symptomscleared within 3 groundmaterial stained light green. days.The wife'srespiratory symptoms remained In intestinalsections of duodenum,mild to at a reducedlevel. moderateinfiltration of enterocyteswith coccidia organismswas presentin each group of DISCUSSION birds. Sectionsof liver,trachea, kidney, heart and At initialsubmission, the clinicalhistory of skeletalmuscle were unremarkable. thesebirds included watery eyes, conjunctivitis, Immunohistochemistry. Lung tissues nasal discharge,and sinusitis,along with his- werestained with a polyclonalantibody specific topathologicobservations of moderateto severe for C. psittaci (Chemicon, Temecula, CA). Tis- rhinitiswith mild lung lesionsthat were highly sue sections were treated with hydrogen per- suggestiveof Mycoplasmainfection (1,4). How- oxide and digested with a proteolytic enzyme. ever,negative polymerase chain reactionDNA After sections were thoroughly rinsed in water probe tests for Mycoplasmagenus ruled out and Triton X Tris-buffered saline, the slides theirpossible role in theseinfections. Thereaf- were transferred to an automated capillary gap ter,immunohistochemistry revealed positive re- system (Ventana Medical Systems, Inc., Tucson, actions for C psittaci in the lungs, and special AZ). Sections were incubated with a serum-free stains for Chlamydiawere ordered. When his- protein-blocking agent (DAKO Corporation, topathologic examination demonstrated Chla- Carpentenia, CA) and incubated with the pri- mydia organisms, the owners were informed of mary antibody. Bound antibodies were detected the diagnosis. Treatment of the birds was im- 802 D. H. Erbeckand S. A. Nunn mediatelyinitiated. Recovery of the originaltis- fection. Chlamydia are very susceptible to sue revealedit to be unsuitablefor isolationat- chemicals that affect their lipid content or the tempts. integrity of their cell walls (1). Even when Once treatment was started, subsequent at- Chlamydia are in tissue debris, their infectivity tempts to isolate and demonstrate Chlamydia is destroyed within minutes by all common dis- organisms were negative, even in birds continu- infectants except cresol and lime (1). Multipli- ing to displaytypical signs. cation of all strains of Chlamydia is strongly Owner discussion with personnel from the inhibited by appropriate concentrations of tet- quailbreeder flocks revealed no unusualdisease racyclines (1). Treatment of Chlamydia with problems that would indicate chlamydiosis. chlortetracycline together with management Subsequent testing of hens from the chukar sanitation to inhibit Pseudomonaslikely contrib- breedersfailed to revealevidence of C psittaci. uted to success in reducing mortality. Coccidi- Although the eggs could have harbored C. psit- osis treatment with monensin reduced stress. taci organisms from external fecal contamina- The origin of the C. psittaci infection was tion, we believe this was unlikely until after the never established. On the affected farm, a hu- first group of chicks was severely affected. man-to-bird zoonotic infection was suspected. Although chlamydiosis can be a serious dis- The adult daughter, who worked in an estab- ease in chukar partridge (12), it has not been lishment where a client regularly brought a pet reported as causing excessive mortality in bob- parrot, was the first to display upper respiratory white quail (Schwartz, pers. comm.). The close symptoms. Others in the family then contract- proximity of each species during the early grow- ed "flulike" symptoms. The epornitic in the ing period suggests that both quail and par- birds was believed to occur after the humans tridge were infected with a pathogenic strain of became ill. Unfortunately, circumstances did C. psittaci that contributed to excess mortality. not allow testing of the parrot. Parrot-to-hu- The clinical signs of chlamydiosis vary great- man-to-chukar transmission is speculative but ly in severity and depend on the age of the bird remains the suspected primary route of trans- and strain of the organism (1). Many avian spe- mission. cies harbor Chlamydia organisms without show- Reservoirs of Chlamydia have been reported ing clinical signs (1). These birds act as carriers to include pigeons, blackbirds, grackles, house and can spread the disease. During periods of sparrows, and killdeer (1). All of these avian stress, carrier birds can break out with chlamy- species could be found in the vicinity of the diosis (11,13). Chlamydiosis is usually systemic; affected farm. Possibly a wild avian species thus, infected birds may be further stressed by transmitted the organism to chukar and bob- Chlamydia infection, and these stressed birds white on the farm. If the C psittaci infection would be susceptible to secondary bacterial in- came from birds, the chukar partridge were fections, including infection with Pseudomonas likely infected first (13). Vertical transmission organisms. Coinfection of C. psittaci and P ae- has been suggested to occur from infected eggs ruginosa likely contributed to the high mortality (5,6,14). Two of six chukar submitted for ex- of the quail and partridge chicks. Coccidia in- amination displayed mild nasal and conjuncti- fection could have contributed to chick stress. val signs. Commingling of the partridge and The mild to moderate histopathologic lesions quail likely resulted in the infection of the bob- suggested that this variable may also have been white chicks. a coinfection factor. This case report was interesting in that it was In chickens and turkeys, Pseudomonas pro- the first reported incident of C. psittaci epor- duces respiratory infections (2). Because P ae- nitic with increased mortality in bobwhite ruginosa is an opportunist in weakened tissues quail. The report also was a reminder of the (3), its isolation from the lungs of the birds was zoonotic potential of chlamydiosis in game not surprising. Pseudomonas aeruginosa was later birds and their handlers. isolated from water lines in the starter facility. Vigorous cleaning and disinfection of water REFERENCES lines and cleaning and sanitation of the brooder and starter building were undertaken and prob- 1. Andersen, A. A., J. E. Grimes, and P. B. ably accounted for less secondary bacterial in- Wyrick. Chlamydiosis (psittacosis, ornithosis). In: Chlamydiosisin bobwhiteand chukar 803

Diseases of poultry, 10th ed. B. W. Calnek, ed. Iowa 8. Morange,A. De la. Psittacoseouinfection spe- State UniversityPress, Ames, IA. pp. 333-349. 1997. ciale determineepar des parruches.M.Sc. Thesis. Ac- 2. Barnes,H. J. Other bacterialdiseases. In: Dis- ademia de Paris, Paris, France. 1895. eases of poultry, 10th ed. B. W. Calnek, ed. Iowa 9. Page, L. A., and J. E. Grimes. Avian chlamy- State UniversityPress, Ames, IA. pp. 289-294. 1997. diosis (ornithosis).In: Diseasesof poultry,8th ed. M. 3. Carter,G. R., M. M. Chengappa,and A. W. S. Hofstad, H. J. Barnes,B. W. Calnek, W. M. Reid, Roberts.Essentials of veterinarymicrobiology, 5th ed. and H. W. Yoder,Jr., eds. Iowa StateUniversity Press, Williams and Wilkins, Baltimore,MD. 1995. Ames, IA. pp. 283-308. 1984. 4. Ley, D. H., and H. W. Yoder. Mycoplasma 10. Schwartz, L. D. Chlamydiosis. In: Grower's gallisepticuminfection. In: Diseases of poultry, 10th referenceon gamebirdhealth. Avicon, Inc., Okimos, ed. B. W. Calnek, ed. Iowa State University Press, MI. pp. 50-51. 1995. Ames, IA. pp. 194-207. 1997. 11. Shewen, P. E. Chlamydial infection in ani- 5. Meyer, K. E Phagocytosis and immunity in mals: a review.Can. Vet. J. 21:2-11. 1980. psittacosis. Schweiz. Med. Wochenschr.71:436-438. 12. Uemori, T., K. Asada, I. Kato, and R. Hara- 1941. sawa. Sensitive detection of Mycoplasmain cell cul- 6. Meyer, K. E, and B. Eddie. Ecology of avian tures by using two step polymerase chain reaction psittacosis, particularlyin parakeets.In: Progressin system. Appl. Microbiol. 15:181-186. 1992. psittacosis research and control, R. Beaudette, ed. 13. Woodard, A. E. Raising chukar partridges. Rutgers University Press, New Brunswick, NY. pp. CooperativeExtension, Universityof California,Da- 52-56. 1958. vis, CA. Leaflet21321. 1982. 7. Mohan, R. Epidemiologicand laboratoryob- 14. Wyrick, P. B., S. Richmond, and B. Chir. Bi- servations of Chlamydia psittaci infection in pet ology of Chlamydiae.Reports from the Symposium birds. J. Am. Vet. Med. Assoc. 184:1372-1374. on Avian Chlamydiosis.J. Am. Vet. Med. Assoc. 195: 1984. 1507-1512. 1989.