Genetic variation in resistance of turkeys to experimental infection with Newcastle disease virus.

Seven hundred eighty male and female turkeys representing four genetic lines were challenged in four experiments with the Texas GB strain of Newcastle disease virus (NDV). The lines of turkeys included two randombred control lines (RBC1 and RBC2), a subline (E) of RBC1 selected for increased egg production, and a subline (F) of RBC2 selected for increased 16-week body weight. Mortality in turkeys of subline F (32.5%) was significantly higher than that in turkeys of line RBC2 (15.8%), subline E (17.5%), and line RBC1 (18.4%). At the end of each experiment, surviving birds were tested for antibody to NDV using the enzyme-linked immunosorbent assay (ELISA) and hemagglutination-inhibition (HI) test. Turkeys of subline E and line RBC1 had significantly lower ELISA antibody titers than those of subline F and line RBC2. Subline F had the highest HI antibody titers, followed in decreasing order by lines RBC2 and RBC1 and subline E. No apparent correlation was found between antibody response and mortality after NDV challenge.     

Molecular epidemiology of Pasteurella multocida in turkeys.

Pasteurella multocida isolated from turkeys during an outbreak of fowl cholera was characterized by serotype and heterogeneity of genes encoding rRNA (ribotype) to investigate the epidemiology of the organism. Isolates were collected between October 1985 and July 1986. The M9 or Clemson University fowl cholera vaccine-like strain was detected in 17% of the flocks with fowl cholera. One particular strain, isolated only from breeder flocks, was recovered from 7 of the 10 breeder flocks examined in this study. Intracompany transmission appeared to be common, implying a failure in biosecurity. Circumstantial evidence indicated that in the field; the incubation period of P multocida in a turkey flock may be between 2 to 7 weeks. Wildlife did not appear to be an important reservoir of P multocida for turkeys during this study period. Ribotyping results tended to discount several of the possible interflock transmissions, as suggested by examination of serotyping results alone; however, serotyping in combination with ribotyping proved helpful in understanding the epidemiology of P multocida in turkeys.     

Pulmonary response to intratracheal challenge with Pasteurella haemolytica and Pasteurella multocida.

Calves were inoculated intratracheally with 5 X 10(7), 5 X 10(8), or 5 X 10(9) colony forming units of either 18-hour stationary phase cultures or 4-hour log phase cultures of Pasteurella haemolytica. The log phase culture at all concentrations produced more severe clinical signs, hematological changes and pulmonary lesions at postmortem examination than did the corresponding stationary phase culture. More severe effects were seen with the larger doses especially with the log phase culture. Fibrinous bronchopneumonia with focal or multifocal necrosis was consistently produced by both the stationary and log phase cultures. To determine if this lesion was peculiar to P. haemolytica or whether it could be produced generally by rapidly growing Gram negative organisms, a 4-hour log phase culture of Pasteurella multocida was prepared in an identical manner to that used for the culture of P. haemolytica and given to calves intratracheally at the high bacterial dose (5 X 10(9]. The P. haemolytica produced more severe clinical, hematological and morphological changes than did the P. multocida. The lesions observed with P. multocida differed morphologically from those of P. haemolytica; there was a suppurative exudative component and minimal to no necrosis with P. multocida. It appears that an important pathogenic principle is produced by the rapidly growing P. haemolytica that causes it to produce a more severe clinical disease and more necrotizing pulmonary lesions than P. multocida.     

Acute airsacculitis in turkeys inoculated with Pasteurella multocida

Thirty female turkeys, inoculated into the caudal thoracic air sacs with Pasteurella multocida were examined from 0 to 6 hours post-inoculation (PI). The air sac reacted rapidly and intensely with exudation of heterophils. Circulating leukocyte and thrombocyte numbers remained normal except for an absolute lymphopenia by 6 hours PI. P. multocida was initially isolated from blood at 3 hours PI. Total cell counts increased markedly in air sac lavage fluids by 1.5 hours PI and continued to increase until 6 hours PI. Heterophils predominated in lavage fluids (greater than 94%), with macrophages comprising the remaining cells. Microscopically occasional heterophils were present within air sac blood vessels and perivascularly by 0.5 hour PI. They became more numerous by 1.5 and 3 hours PI when transepithelial migration into the air sac lumen was seen. By 6 hours PI, there was diffuse, severe swelling of air sac epithelium and mesothelium, and bacteria were located in air sac interstitium. Ultrastructurally, endothelial and air sac epithelial cells were swollen and vacuolated Interdigitating processes of air sac epithelial cells were separated. These results indicate that air sacs can be the portal of entry for P. multocida into the systemic circulation, probably via damaged air sac epithelium.     

Serum resistance as an indicator of virulence of Pasteurella multocida for turkeys

Wildlife isolates of Pasteurella multocida, whose virulence for turkeys had previously been determined by intravenous inoculation, were characterized regarding their ability to survive incubation in fresh non-immune turkey serum. The relative virulence of the isolates was significantly associated with their ability to resist the bactericidal power of the serum as determined by standard plate counts following incubation. Organisms with a high survival value were more virulent; those with a low survival value were less virulent. A statistical model was specified and was successfully used to predict relative virulence of the P. multocida isolates. This method of assaying serum resistance was rapid, repeatable, and practical and could be performed with minimal laboratory equipment. Also studied was the serum resistance of seven serotype 3, 4 isolates obtained from the lungs of M9-vaccinated turkeys from seven flocks experiencing increased mortality due to fowl cholera. These isolates were shown to be identical to the M9 vaccine by restriction endonuclease analysis of chromosomal DNA. Six of the seven isolates had higher serum survival values than the original M9 vaccine.     

Comparative susceptibility of caponized and uncaponized tom turkeys to Pasteurella multocida.

When susceptibility to virulent Pasteurella multocida was compared, there was no significant (P greater than 0.05) difference between caponized and uncaponized tom turkeys. Neither was there any significant (P greater than 0.05) difference between the surviving caponized and uncaponized toms in the development of serum anti-P. multocida antibody. However, at 28 weeks of age, the average live body weight of the caponized toms was significantly (P less than 0.05) lower than that of the uncaponized toms. Turkeys were caponized when 9 weeks old, and different groups were exposed to P. multocida when 13, 18, 23, and 28 weeks old.     

Vaccination of turkeys with cell-free culture filtrate of Pasteurella multocida

Turkeys given cell-free culture filtrate (CCF) of Pasteurella multocida strain R44/6 orally, via air sacs, or subcutaneously mixed 1:1 with incomplete Freund's adjuvant (IFA) at 6 and 9.5 weeks of age were compared with negative controls given bacteriologic medium and positive controls vaccinated with a commercial bacterin. At 13 weeks of age, serum antibody titers to P. multocida were detectable only in turkeys given CCF in IFA (low titers) and positive control turkeys (high titers), at which time turkeys were challenged orally with either the homologous strain or strain P-1059. Protection against challenge with strain R44/6 was provided by the commercial bacterin, CCF in IFA, and CCF given via air sacs. When turkeys were challenged with strain P-1059, protection was superior in turkeys given CCF via air sacs, intermediate in turkeys given commercial bacterin or CCF in IFA, and absent in negative control turkeys and turkeys given CCF orally. These results indicate CCF is an effective immunogen when administered via the lower respiratory tract for protecting turkeys against pasteurellosis.     

Genetic variation in response of turkeys to experimental infection with Bordetella avium

One hundred ninety-six male and female turkeys representing two genetic lines were experimentally infected with Bordetella avium. The lines of turkeys included a randombred control line (RBC2) and a subline (F) of RBC2 selected for increased 16-wk body weight. No difference was found between lines RBC2 and F in the number of days to onset of clinical signs, and no mortality due to B. avium infection was observed in either line. Interestingly, however, a significant depression (12%) occurred in body weight of F line poults infected with B. avium, but no significant depression occurred in body weight of RBC2 poults.     

Resistance plasmids of Pasteurella multocida isolated from turkeys

From 1940 through 1978, fifty-eight strains of Pasteurella multocida (serotype 3) were isolated from turkeys throughout the United States and were examined for R-plasmids. Forty-one of the isolates contained plasmid DNA, of which 7 isolates were found to encode resistance to tetracycline, streptomycin, and sulfonamides, or to streptomycin and sulfonamides. The R-plasmids were 2 to 10 megadaltons, nonconjugal, and contained a moles percent guanine plus cytosine ratio in the range of 57 to 61. The R-plasmids did not belong to any of the 19 incompatibility groups evaluated, including Inc Q. Digestion with restriction endonuclease indicated that 2 of the plasmids from P multocida isolated in 1960 and 1962 were identical, whereas 4 of the 5 plasmids obtained from P multocida isolated after 1966 were identical, with the 5th plasmid closely related to the other 4. The results indicated that R-plasmids were not widely dispersed among P multocida (serotype 3) isolated from turkeys in the United States. The nontransmissible nature of these plasmids was probably the major reason for their lack of dissemination.     

Pasteurella multocida recovered from live turkeys: prevalence and virulence in turkeys

Swabs of the oropharynges of 801 live turkeys (621 meat birds and 180 breeders), collected from 15 flocks that had experienced an outbreak of fowl cholera and from 12 non-outbreak flocks, were screened for the presence of Pasteurella multocida. Turkeys from outbreak flocks were sampled within 2 to 9 weeks of the outbreak. Forty-nine isolates of P. multocida were recovered from turkeys in 11 of the outbreak flocks, and none were recovered from turkeys in non-outbreak flocks. Isolation rates varied from 0 to 72% of turkeys sampled in a flock. Nineteen isolates were tested for virulence by injecting them intravenously into turkeys, and 14 were lethal. Results demonstrated that for purposes of disease control, meat birds in fowl-cholera-outbreak flocks should be considered carriers of potentially virulent P. multocida for the life of the flock.     

Prevalence and characteristics of Pasteurella multocida in commercial turkeys

The oropharyngeal regions of 680 meat turkeys and 55 breeder turkeys from nine outbreak farms, three history-outbreak farms, and 19 nonoutbreak farms in Ohio, Indiana, and Pennsylvania were cultured to determine the prevalence of Pasteurella multocida in turkeys. Pasteurella multocida was recovered from 32 out of 105 turkeys belonging to outbreak farms. Pasteurella multocida was not recovered from either history-outbreak or nonoutbreak farms. Characterization via capsular and somatic serotyping, biotyping, restriction endonuclease analysis, and antimicrobial susceptibility testing was performed on all recovered P. multocida isolates. Pasteurella multocida serotype A:1 and somatic serotype 1 with an un-typable capsular serogroup (UT:1) were the most common serogroups found. All isolates belonged to biotype P. multocida ssp. multocida. EcoRI, HpaII, and HindIII restriction enzyme digestions identified three, five, and five restriction fragment length polymorphism profiles, respectively. A majority of the isolates were susceptible to amikacin, ampicillin, ceftiofur, cephalothin, enrofloxacin, florfenicol, gentamicin, neomycin, novobiocin, oxacillin with 2% NaCl, sarafloxacin, tilmicosin, and trimethoprim with sulphadiazine and resistant to clindamicin, penicillin, tiamulin, and tylosin.     

Dermal necrosis caused by Pasteurella multocida infection in turkeys

Severe dermal necrosis caused by Pasteurella multocida Serotype 1 was diagnosed in three dressed turkey carcasses and two live turkeys from a commercial flock. The dressed carcasses were among several condemned at a processing plant. The isolate, P. multocida Serotype 1, produced progressive dermal necrosis when experimentally inoculated into injured skin of turkeys. The organism was reisolated from the dermal lesions. The turkey houses were found to be infested by mice; the skin injury and infection with P. multocida probably originated from mouse bites.     

Immune defence mechanism against blood-borne Pasteurella multocida in turkeys

Humoral and cellular immune defence factors involved in controlling blood-borne Pasteurella multocida were investigated in turkeys by the passive transfer of immune serum or by the treatment with macrophage-activating agents. The treated and untreated birds were intravenously inoculated with a virulent strain of P multocida, and the viable bacteria in the blood, liver and spleen were counted. In untreated birds, the bacteria were rapidly removed from the blood, and the majority were recovered from the liver and spleen 120 minutes after inoculation. Neither the transfer of immune serum nor the treatment with macrophage-activating agents significantly influenced the clearance rate of bacteria from the blood. The number of bacteria recovered from the liver 120 minutes after inoculation was slightly lower in the birds treated with macrophage-activating agents and significantly lower in those given immune serum than in the untreated birds. None of the treatments, however, significantly changed the number of bacteria recovered from the spleen 120 minutes after inoculation. The results suggest that the phagocytes in the liver, but not in the spleen, play a crucial role in the intravascular defence against P multocida in the presence of specific antibodies.     

Spondylitis in turkeys associated with experimental Pasteurella anatipestifer infection.

Nine previously vaccinated turkeys were inoculated intravenously with Pasteurella anatipestifer, and blood samples were taken periodically to evaluate the potential of chronically infected turkeys to serve as reservoirs of infection for blood-feeding arthropod vectors. Vertebral osteomyelitis (spondylitis), as yet unreported in the literature in association with infection with the organism, was found in the thoracic vertebrae of five out of nine inoculated turkeys, and P. anatipestifer was isolated from the thoracic vertebrae of three of the five. The organism was isolated from the peripheral blood of six turkeys 24 hours postinoculation and from the peripheral blood of one turkey 7 days postinoculation. The organism was also isolated from the heart blood of two birds at necropsy--from one at 21 days and, following an intramuscular injection of dexamethasone, from the other turkey at 38 days postinoculation.     

Acute airsacculitis in turkeys inoculated with cell-free culture filtrate of Pasteurella multocida

Twenty-six female and 26 male turkeys, inoculated into the caudal thoracic air sacs with cell-free culture filtrate of Pasteurella multocida strain R44/6, were examined from 0 to 6 hours post-inoculation and compared with 26 female and 26 male sham-inoculated control turkeys given brain-heart-infusion broth. The air sac reacted rapidly with exudation of heterophils. Microscopically, low numbers of heterophils were present within air sac blood vessels and also perivascularly by 0.5 hour after inoculation. These became more numerous by 1.5 and 3 hours post-inoculation. By 6 hours post-inoculation, there was severe swelling of air sac epithelial and mesothelial cells and thickening of the air sac by proteinaceous fluid and heterophils. Ultrastructurally, mesothelial and air sac epithelial cells were vacuolated, and interdigitating processes of epithelial cells were separated. Microscopically, in control turkeys, rare heterophils were present perivascularly at 1.5, 3, and 6 hours after inoculation. Ultrastructurally, all features were normal. In turkeys given cell-free culture filtrate, total cell counts in air sac lavage fluids increased markedly by 3 hours post-inoculation in which heterophils predominated (greater than 97%). There were only slight increases in cell counts of air sac lavages from control turkeys. The circulating blood heterophil cell count dropped transiently at 1.5 hours post-inoculation, followed by a return to normal 3 hours after inoculation, and by heterophilia by 6 hours post-inoculation in turkeys given either cell-free culture filtrate or brain-heart-infusion broth. These results indicate cell-free culture filtrate of P. multocida induces hematologic, cytologic, and morphologic changes indistinguishable from those induced by cultures of P. multocida.     

Safety testing of Pasteurella multocida vaccines and bacterins in turkeys

When U.S. Department of Agriculture-licensed Pasteurella multocida vaccines and bacterins were administered to healthy turkeys under controlled laboratory conditions, they did not cause an increase in death loss.     

Association of complement sensitivity with virulence of Pasteurella multocida isolated from turkeys

Two strains of Pasteurella multocida, both derivatives of strain P1059, were compared for virulence for 14-week-old turkeys and sensitivity to turkey plasma. Strain P1059-1, a nalidixic-acid-resistant mutant of P1059 with an LD50 of approximately 10(3) colony-forming units (CFU), was more resistant to the bactericidal effects of fresh turkey plasma at 37 C than avirulent strain P1059-1A. P1059-1A, with an LD50 of approximately 10(8) CFU, is an acapsular variant of P1059-1 that spontaneously arose after prolonged passage on artificial medium. The bactericidal effect on P1059-1A was removed when turkey plasma was treated with heat or with zymosan, maneuvers that removed hemolytic complement activity from turkey plasma.