Turkey coryza: toxin production by Bordetella avium

Twelve strains of Bordetella avium representing isolates from turkeys in the United States, the Federal Republic of Germany, and the Republic of South Africa were tested for toxin production. Sterile filtered sonicates from 9 of 12 strains contained a toxin that was lethal for 7-to-10-day-old poults. Mice were also susceptible to the lethal effects of the toxin. No differences in susceptibility to the toxin were found between Beltsville small white and broad-breasted white poults. The toxin was solubilized by sonication and inactivated by heating at 56 C for 30 min. Treatment with formalin or proteolytic enzymes inactivated the toxin, indicating that it is probably a protein. The evidence suggests that the toxin is involved in the pathogenesis of turkey coryza.     

The effects of heat-labile Bordetella avium toxin on turkey poults

Intraperitoneal exposure of turkey poults to various concentrations of Bordetella avium sonicate containing heat-labile toxin indicated a high degree of toxicity: poults died after exposure to sonicate containing as little as 6.32 micrograms of protein. The toxic effects were dose-related: poults that received sonicate containing 158 micrograms of protein died in 4 to 6 1/2 hours, those that received 31.6 and 6.32 micrograms of protein died in 25 to 30 hours, and those that received 1.2 micrograms survived the 6-day course of the study and were apparently unaffected. Histologic examination of poults that received lethal doses of the toxin revealed degeneration and necrosis of parenchymal cells of the liver and pancreas as well as hyperemia, hemorrhage, and necrosis of the mucosa of the small intestine. No lesions were observed in poults that received the sublethal dose of toxin or in unexposed poults. Repeated intranasal exposure of poults to the sonicate did not produce clinical signs of disease or gross lesions.     

Turkey coryza: selected tests for detection and neutralization of Bordetella avium heat-labile toxin

Bordetella avium heat-labile toxin (HLT) was lethal for poults, mice, and embryonating chicken eggs. It produced hemorrhagic lesions in turkey and guinea pig skin. Antiserum made in turkeys neutralized the lethality of the toxin and its ability to produce hemorrhagic skin lesions. Further, antiserum against HLT of an Ohio strain neutralized lethality of HLT of strains from Iowa, North Carolina, and West Germany. The antiserum did not neutralize lethality of HLT from B. bronchiseptica. Bordetella avium HLT was not ciliostatic for turkey tracheal-ring cultures and did not stimulate adenyl cyclase activity using mouse adrenal cell cultures.     

Effects of Bordetella avium toxin on turkey tracheal organ cultures as measured with a tetrazolium-reduction assay

Turkey tracheal organ cultures (TOCs) were exposed to one of the following Bordetella avium fractions or controls: live B. avium, formalin-killed B. avium, B. avium sonicate, heat-inactivated sonicate, culture supernatant, heat-inactivated culture supernatant, phosphate-buffered saline, or brain-heart infusion broth. After the TOCs were incubated for 2 hours with the bacterial fractions, the cellular metabolism of each TOC was evaluated using a tetrazolium chloride reduction assay, and cellular morphology was determined by light microscopy. Additionally, bacterial fractions and controls were injected into turkeys to test lethality. Although the bacterial sonicate was lethal for turkeys, neither the sonicate nor any other B. avium fraction significantly affected the metabolism or morphology of turkey TOCs.     

Brain monoamine concentrations in turkey poults infected with Bordetella avium

Six hours post-hatch, large white turkey poults were inoculated intranasally with 5 X 10(7) organisms of the "W" isolate of Bordetella avium. Three hours after inoculation and subsequently on days 7, 10, 14, 21, and 28 postinoculation, 30 infected and 30 uninfected control poults were injected intramuscularly with alpha methylparatyrosine (AMPT, 250 mg/kg), and an additional 30 of each group received a saline vehicle. Three hours after AMPT injection, whole brains from treated and untreated poults from both infected and control groups were removed, weighed, and frozen until assayed for norepinephrine (NE), dopamine (DA), and serotonin (5-HT). B. avium-infected poults had a significant reduction in steady-state NE and Da and a greater depletion of NE and DA after AMPT treatment compared with control poults. Infected poults had significantly reduced whole brain 5-HT concentrations, which persisted through 21 days postinoculation. Altered brain NE, DA, and 5-HT concentrations suggest that the B. avium-infected poults may be less able to cope with additional stressors.     

Turkey coryza: lack of correlation between plasmids and pathogenicity of Bordetella avium

Plasmids were removed from pathogenic Bordetella avium using a variety of treatments. The plasmid-cure rates depended on the treatment and isolate. Pathogenicity of B. avium in turkey poults was not altered by removal of plasmids.     

Protection of turkey poults from Bordetella avium infection and disease by pili and bacterins

The role of pili in protection against Bordetella avium infection in turkey poults was studied. An isolate that produced the largest number of pili under growth conditions developed in our laboratory was used for preparation of pili and bacterin and for challenge. The pili were isolated, purified, examined by electron microscopy, and tested for purity by gel electrophoresis. Poults were vaccinated with oil-adjuvant pili, formaldehyde- or merthiolate-inactivated bacterins, or a commercial bacterin. Poults were vaccinated once or twice subcutaneously at different ages and challenged intranasally with a pathogenic B. avium isolate 5 days following the last vaccination. A few vaccinated birds had very mild clinical signs. B. avium was isolated from the sinuses of a few vaccinated birds, and growth was scanty. The mean colony counts from tracheal sections was significantly higher (P less than 0.1) in unvaccinated challenged poults than in vaccinated challenged poults. It is postulated that B. avium pili are important immunogens in turkey poults.     

Clinical outbreak of Bordetella avium infection in two turkey breeder flocks

An acute upper respiratory disease was observed in two broad-breasted white (BBW) turkey primary breeder flocks. Associated clinical signs included sneezing, depression, and a deep dry cough originating from large conducting airways. Morbidity reached approximately 15-20% of the hens in an affected house. None of the turkeys died, and total feed consumption was not affected. A minimal effect upon egg production was noticed. Sera from an acutely affected flock exhibited a marked rise in titer to Bordetella avium compared with preinfection sera samples. In Case 1, B. avium was isolated in pure culture from affected birds. In Case 2, B. avium was diagnosed by serological results and clinical signs; bacteriological examination was not attempted. The findings presented here are consistent with an acute clinical outbreak of B. avium-induced turkey rhinotracheitis (turkey coryza) in BBW turkey breeder hens.     

Effect of Bordetella avium infection on electrocardiograms in turkey poults.

Electrocardiograms (ECGs) were recorded from turkey poults infected with the W isolate of Bordetella avium. Strip chart data (amplitudes and intervals) were measured at 7, 14, 21, and 28 days postinoculation and compared with data from uninoculated controls. B. avium infection altered P-, RS-, and T-wave amplitudes, as well as P-R and S-T intervals throughout the 4-week experimental period. Heart rates were similar over the 4-week period in control poults, while rates in the infected birds were variable. Alterations in ECGs associated with B. avium infection appeared to be the result of disturbances in the cardiovascular system and may be associated with the pathogenicity of the organism in turkey coryza.     

Prevalence,virulence attributes,and antibiogram of Bordetella avium isolated from turkeys in Egypt

Tropical Animal Health and Production - Turkey coryza is a major respiratory disease caused by Bordetella avium (B. avium). It occurs in all ages of turkeys and is characterized by high morbidity...     

Effects of Bordetella avium on lymphoid tissue monoamine concentrations in turkey poults

Six hours post-hatch, large white turkey poults were inoculated intranasally with 5 x 10(7) cells of the W isolate of Bordetella avium. Three hours after inoculation and subsequently on days 7, 10, 14, 21, and 28 postinoculation, poults from infected and control groups were killed by cervical dislocation. Thymus, spleen, and bursa of Fabricius were removed, weighed, and frozen until assayed for norepinephrine (NE), dopamine (DA), and serotonin (5HT). B. avium infection caused a reduced concentration of NE, DA, and 5HT in the spleen, thymus, and bursa of Fabricius. The reduced concentrations of these monoamines in lymphoid tissues of diseased poults may be a normal response during the course of a disease or during the mounting of an immune response.     

Influence of Bordetella avium infection on association of Escherichia coli with turkey trachea

Four-week-old Bordetella avium-infected and B avium-free turkeys were inoculated intratracheally with a suspension of fimbriated or nonfimbriated Escherichia coli. Numbers of E coli associated with tracheal sections were determined at postinoculation hour (PIH) 1 or 6. Significantly (P less than 0.05) greater numbers of E coli were isolated from the tracheas of B avium-infected turkeys compared with numbers in B avium-free turkeys. In B avium-free turkeys, tracheal associated E coli were 90% less at PIH 6 compared with that at PIH 1. However, in B avium-infected turkeys, numbers of E coli were not affected by postinoculation time. Seemingly, B avium-infected turkeys had reduced capacity to clear E coli from the trachea.     

Adherence of Bordetella avium to turkey tracheal mucosa: effects of culture conditions

Adherence of Bordetella avium to the tracheal mucosa of turkeys was evaluated, using bacteria grown under different culture conditions. Several solid and liquid media were used at incubation times of 12, 24, 36, or 48 hours with incubation temperatures of 18, 26.5, or 35 C. Adherence of B avium was greatest when the bacteria were grown on solid media at 35 C. Use of Bordet-Gengou or brain-heart infusion agar was associated with significantly greater (P less than 0.05) adherence compared with adherence of bacteria grown on other media. Adherence was greatest, using cultures in the stationary phase of growth; however, with some media, adherence diminished when incubation was extended beyond 36 hours. Adherence of B avium was reduced but not completely prevented when cultures were incubated at 18 C.     

Isolation of Bordetella avium from poultry

Four Australian isolates of Gram-negative nonfermentative bacteria obtained from poultry were compared with reference strains of Bordetella avium, Bordetella bronchiseptica and Alcaligenes faecalis. The Australian isolates were identified as Bordetella avium. A routine procedure for the identification of this recently recognised poultry pathogen is described.     

Contribution to the taxonomy of the turkey coryza agent: cellular fatty acid analysis of the bacterium

Gas-liquid chromatography was used to analyze bacterial cellular fatty acids to elucidate the relatedness of the turkey coryza (TC) bacterium to Alcaligenes spp., Bordetella spp., and other gram-negative bacteria. The results indicated that the TC bacterium is not closely related to Alcaligenes faecalis or any of the reference strains of Alcaligenes and Bordetella studied. Most urease-positive bacterial isolates obtained from the upper respiratory tract of turkeys were identified as Bordetella bronchiseptica. It is suggested that Bordetella avium is a suitable designation for the TC bacterium formally called Bordetella-"like" and A. faecalis type I. It is also suggested that the nonpathogenic bacterium previously identified as type II A. faecalis be designated B. avium-like until further taxonomic studies are available. Furthermore, it is proposed that the term turkey coryza be used to refer to the disease induced by this bacterium.     

Lack of protection against Bordetella avium in turkey poults exposed to B. avium-like bacteria

Six laboratory experiments were designed to determine whether poults infected with the nonpathogenic Bordetella avium-like (BAL) bacteria would develop immunity to B. avium (BA), the causative agent of turkey coryza. The BAL bacteria were isolated from poults given that organism, but few colonies were observed by 3 weeks postexposure. No serum-agglutinating antibody to the BAL bacterium was detected in poults exposed to that organism. Poults exposed to BAL bacteria either once or twice at different ages were not protected from infection or disease following experimental challenge between 1 and 7 weeks of age with pathogenic BA.     

An enzyme-linked immunosorbent assay for detection of Bordetella avium infection in turkey flocks: sensitivity, specificity, and reproducibility.

An enzyme-linked immunosorbent assay (ELISA) for detection of Bordetella avium infection in turkey poults was developed. One-week-old poults challenged intratracheally with 10(12) colony-forming units of B. avium had detectable titers (greater than or equal to 11), with an average of 13.6% positive samples when the birds were 6 to 11 weeks old. The method was sensitive enough to detect maternal antibodies to B. avium in poults up to 3 weeks of age. The same poults challenged at 1 week of age had 100% tracheal infection up to 3 weeks of age, which dropped to 0% by 6 weeks. The method resulted in no false-positive samples (titer = 0) from birds not infected with B. avium and tested weekly between 4 and 11 weeks of age. Antibodies in turkey flocks infected with Newcastle disease virus, hemorrhagic enteritis virus, and Mycoplasma meleagridis, and birds infected with Escherichia coli had no apparent cross-reactivity to the B. avium antigens used in the ELISA. The percentages of B. avium-positive serum samples collected from different turkey flocks did not significantly differ (P greater than 0.05) when samples were tested by the developed ELISA at different times, an indication of the reproducibility of the method.     

Tracheal cilia response to exogenous niacin in drinking water of turkey poults infected with Bordetella avium.

Niacin was added daily to the drinking water of control and Bordetella avium-infected turkey poults at a dosage of 0, 70, and 280 mg/liter over a 2-week experimental period. Fourteen days postinoculation, tracheal sections were examined by histological and morphometrical analysis of cilia, as well as agar plate isolation for bacteria. Infected poults exhibited a 96-97% loss of cilia along the tracheal epithelial border, compared with only a 4-5% loss in controls. Infected poults receiving niacin in the drinking water exhibited only a 61.0% and 76.0% loss of cilia at doses of 70 mg/liter and 280 mg/liter, respectively. The results indicate that niacin treatment may influence the pathogenicity of B. avium infection.     

Effect of fasting on plasma thyroid and adrenal hormone levels in turkey poults infected with Bordetella avium.

Plasma triiodothyronine (T3), thyroxine (T4), and corticosterone (CS) were measured in fasted and nonfasted control and Bordetella avium-infected poults. The stress of B. avium infection increased plasma CS, and fasting for 24 hours caused a further significant increase in CS levels. Plasma T3 was not affected by the infection, but fasting caused a significant reduction in both control and infected poults. Plasma T4 of fasted poults was increased in both control and infected groups, but infection attenuated the plasma T4 response. Total iodothyronines were increased in the control-fasted poults more than in infected-fasted poults, indicating a reduced responsiveness to stress by the thyroids of infected poults. Changes in plasma thyroid hormones and CS partially explain the decreased body weight gain and decreased body temperature after exposure to B. avium.     

Further characterization of the agent causing coryza in turkeys

A total of 128 isolates of Alcaligenes faecalis, from the respiratory tract of turkeys and chickens, were identified and divided into two types designated type I and type II. Type I isolates were pathogenic in poults, hemagglutinated guinea pig red blood cells (RBCs), and did not grow on minimal essential medium (MEM) agar, and most did not grow in 6.5% NaCl broth. Type II isolates were nonpathogenic and nonhemagglutinating and grew on MEM agar, and most grew in 6.5% NaCl broth. Hemagglutination of guinea pig RBCs was a reliable characteristic for distinguishing type I from type II isolates, and it correlated with pathogenicity. In serological studies using 62 type I and 21 type II isolates, cross-reactions were observed when type I but not type II antigens were used to test antisera in the microagglutination test. Eleven bacterial isolates, different from type I and type II isolates, were urease-positive. Although frequently isolated from turkeys with coryza, these isolates were nonpathogenic and were always found in association with type I A. faecalis. Urease-positive isolates and type I and type II A. faecalis isolates were stable following 50 in vitro passages. Bordetella avium sp. nov. (the nomenclature suggested in Europe for A. faecalis) was pathogenic in poults. The colonial morphology, biochemical characteristics, and hemagglutinating activity of B. avium sp. nov. were the same as those of type I A. faecalis isolates. Based on the results of these studies, it was concluded that type I A. faecalis is the etiologic agent of turkey coryza.